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1

Thyroid Hormone Regulates the Cell Cycle Inhibitor p27Kip1 in Postnatal Murine Sertoli Cells  

Microsoft Academic Search

Thyroid hormone regulates early postnatal Sertoli cell pro- liferation. Transient neonatal hypothyroidism allows pro- longed postnatal Sertoli cell mitogenesis and doubles adult Sertoli cell numbers, testis weight, and sperm production. The mechanism of this effect is unknown. Cell proliferation is stimulated by cyclins and cyclin-dependent kinases and in- hibited by cyclin-dependent kinase inhibitors (CDKIs). T3 reg- ulates the CDKI p27Kip1

DENISE R. HOLSBERGER; SIWANON JIRAWATNOTAI; HIROAKI KIYOKAWA; PAUL S. COOKE

2003-01-01

2

Sertoli cells as biochambers  

NASA Technical Reports Server (NTRS)

According to the present invention, there is provided a biological chamber system having a biochamber defined by outer walls of Sertoli cells. Also provided is a transplantation facilitator including a biochamber. A method of making biochambers by co-culturing facilitator cells and therapeutic cells and then aggregating the facilitator celes is also provided. Also provided is a method of transplanting cells by incorporating transplant cells into a biochamber and transplanting the biochamber containing the transplant cells.

Cameron, Don F. (Inventor); Sanberg, Paul R. (Inventor); Saporta, Samuel (Inventor); Hushen, Joelle J. (Inventor)

2004-01-01

3

Germ cell binding to rat Sertoli cells in vitro.  

PubMed

The interaction between male germ cells and Sertoli cells was studied in vitro by co-incubation experiments using isolated rat germ cells and primary cultures of Sertoli cells made germ cell-free by the differential sensitivity of germ cells to hypotonic shock. The germ cell/Sertoli cell interaction was examined morphologically with phase-contrast and scanning electron microscopy and then quantified by measuring radioactivity bound to Sertoli cell cultures after co-incubation with added [3H]leucine-labeled germ cells. Germ cell binding to Sertoli cell cultures was the result of specific adhesion between these two cell types, and several features of this specific adhesion were observed. First, germ cells adhered to Sertoli cell cultures under conditions during which spleen cells and red blood cells did not. Second, germ cells had a greater affinity for Sertoli cell cultures than they had for cultures of testicular peritubular cells or cerebellar astrocytes. Third, germ cells fixed with paraformaldehyde adhered to live Sertoli cultures while similarly fixed spleen cells adhered less tightly. Neither live nor paraformaldehyde-fixed germ cells adhered to fixed Sertoli cell cultures. Fourth, germ cell binding to Sertoli cell cultures was not immediate but increased steadily and approached a maximum at 4 h of co-incubation. Saturation of germ cell binding to Sertoli cell cultures occurred when more than 4200 germ cells were added per mm2 of Sertoli cell culture surface. Finally, germ cell binding to Sertoli cell cultures was eliminated when co-incubation was performed on ice. Based on these observations, we concluded that germ cell adhesion to Sertoli cells was specific, temperature-dependent, and required a viable Sertoli cell but not necessarily a viable germ cell. These results have important implications for understanding the complex interaction between Sertoli cells and germ cells within the seminiferous tubule and in the design of future experiments probing details of this interaction. PMID:3442698

DePhilip, R M; Danahey, D G

1987-12-01

4

Sertoli cell plasma membrane polypeptides involved in spermatogenic cell-Sertoli cell adhesion.  

PubMed

This study concerns Sertoli cell-spermatogenic cell adhesive interactions in the seminiferous tubule. Sertoli cell surface polypeptides involved in germ cell-Sertoli cell adhesion were identified by serological inhibition of an in vitro Sertoli-germ cell adhesion assay. This assay was modified from a previously reported adhesion assay, and employs a scanning laser cytometer for quantification of adherent cells. Reactivity of the polyclonal antiserum raised against rat Sertoli cells was also assessed via immunofluorescent microscopy. The addition of antiserum to the adhesion assay resulted in a 42% to 66% inhibition of cell-cell adhesion. Moreover, preincubation of antiserum with Sertoli cell monolayers resulted in a significant reduction of spermatogenic cell binding. Conversely, preincubation of antiserum with germ cells resulted in no reduction. Western blot analysis of the antiserum against purified Sertoli cell membranes indicated reactivity with four polypeptides. The data suggest that one or more of these polypeptides are directly involved in the adhesion of germ cells to Sertoli cell monolayers in vitro. PMID:1597400

Newton, S C; Millette, C F

1992-01-01

5

Giant Sertoli cell nodule of the testis: distinction from other Sertoli cell lesions  

PubMed Central

The case of a 33?year?old man with a clinically suspected testicular neoplasm is reported here. The radical orchidectomy specimen showed a sharply demarcated, firm, yellow–white 1?cm nodule beneath the tunica albuginea at the upper pole. Microscopical examination showed the encapsulated nodule to be composed of tubules lined by immature Sertoli cells with interspersed spermatogonia and an interwoven network of hyalinised basement membrane having foci of calcification. Immunohistochemical studies verified the fetal phenotype of the Sertoli cells and the non?neoplastic nature of the germ cell component. Except for the large size, the findings were identical to those of a Sertoli cell nodule—a typically microscopic, unencapsulated lesion commonly detected in cryptorchid testes. The term “giant Sertoli cell nodule” is used for this unique, hitherto undescribed lesion and its distinction from other Sertoli cell lesions of the testis is considered here.

Barghorn, A; Alioth, H-R; Hailemariam, S; Bannwart, F; Ulbright, T M

2006-01-01

6

N-cadherin mediates Sertoli cell-spermatogenic cell adhesion.  

PubMed

The complex topological association of Sertoli cells and spermatogenic cells in the testis suggests the existence of cell surface adhesion molecules that regulate cellular interactions within the seminiferous epithelium. The recent report of N-cadherin mRNA expression in the mouse testis implies the involvement of this known adhesion molecule in testicular cell binding. Accordingly, here we report that (1) N-cadherin is found on the surface membranes of rat spermatogenic cells and on Sertoli cells, and (2) that N-cadherin is a partial mediator of Sertoli cell-germ cell adhesion as tested in an vitro cell-cell binding assay. Antiserum directed against the N-cadherin cell adhesion recognition sequence was used for Western blot anlaysis of purified plasma membranes from Sertoli cells and from spermatogenic cells. Both membrane preparations exhibited reactivity at an appropriate M(r) of about 130 kDa. In addition, immunofluorescence assays demonstrated that both germ cells and Sertoli cells were labeled by anti-N-cadherin. Finally, the antiserum was included in a cytometer-assisted cell-cell binding test to determine its inhibitory ability. The antiserum consistently reduced specific testicular cell-cell adhesion by 30%-50%. This is the first demonstration that antibodies directed against the cadherin cell adhesion recognition sequence are capable of inhibiting cell-cell interactions. Pre-incubation of either rat Sertoli cells or spermatogenic cells alone was sufficient to achieve statistically significant inhibition of intercellular adhesion. We conclude, therefore, that N-cadherin is expressed by both Sertoli cells and spermatogenic cells and that N-cadherin is one of a number of regulatory molecules mediating local cellular associations in the mammalian seminiferous tubule. PMID:8400407

Newton, S C; Blaschuk, O W; Millette, C F

1993-05-01

7

Reprogramming sertoli cells into pluripotent stem cells.  

PubMed

Abstract Induced pluripotent stem cells (iPSCs) have potential applications in the restoration of fertility, regenerative medicine, and animal biotechnology. In this study, we present the induction of iPSCs from mouse Sertoli cells (SCs) by introducing four factors-Oct4, Sox2, Klf4, and c-Myc. As early as day 3 after induction, expression of these factors was detected and typical embryonic stem-like cells began to form. On day 18, these exogenous genes were silenced and colonies were selected according to morphological characteristics. The iPSCs induced from SCs, termed SCiPSCs, strongly expressed pluripotent markers, showed a normal karyotype, and had proliferation and differentiation characteristics similar to those of embryonic stem cells (ESCs), both in vitro and in vivo. Furthermore, exposure of SCiPSCs to nitric oxide (NO) allowed them to maintain pluripotency through the activation of the pluripotent genes Oct4 and Sox2 and upregulation of Nanog expression. Moreover, NO prevented SCiPSCs from undergoing apoptosis by activating the antiapoptotic genes Bcl2 and Bcl2lll, downregulating the proapoptotic genes Bak1 and Casp7, and blocking the activation of the proapoptotic gene Bac. These effects were reversed by exposure to l-NG-monomethylarginine (l-NMMA), a NO inhibitor. These data demonstrate that iPSCs can be generated from SCs and that the self-renewal and pluripotency of SCiPS cells can be maintained in the presence of NO. PMID:24802333

Sun, Hongyan; Zhang, Guomin; Dong, Fulu; Wang, Feng; Cao, Wenguang

2014-06-01

8

Sertoli cell signaling by Desert hedgehog regulates the male germline  

Microsoft Academic Search

Background: In mammals, testis development is initiated in the embryo in response to the expression of the sex determining gene, Sry, in Sertoli cell precursors. Subsequently, Sertoli cells are thought to play a central role in male-specific cell interactions, including those that occur during spermatogenesis. However, the molecular nature of these interactions is poorly understood. Desert hedgehog (Dhh) encodes a

Mark J Bitgood; Liya Shen; Andrew P McMahon

1996-01-01

9

Purinergic signalling mobilizes mitochondrial Ca2+ in mouse Sertoli cells  

PubMed Central

Abstract Intimate bidirectional communication between Sertoli cells and developing germ cells ensures the integrity and efficiency of spermatogenesis. Yet, a conceptual mechanistic understanding of the physiological principles that underlie Sertoli cell autocrine and paracrine signalling is lacking. Here, we characterize a purinergic Ca2+ signalling network in immature mouse Sertoli cells that consists of both P2X2 and P2Y2 purinoceptor subtypes, the endoplasmic reticulum and, notably, mitochondria. By combining a transgenic mouse model with a dedicated bioluminescence imaging device, we describe a novel method to monitor mitochondrial Ca2+ mobilization in Sertoli cells at subcellular spatial and millisecond temporal resolution. Our data identify mitochondria as essential components of the Sertoli cell signalling ‘toolkit’ that control the shape of purinergic Ca2+ responses, and probably several other paracrine Ca2+-dependent signals.

Veitinger, Sophie; Veitinger, Thomas; Cainarca, Silvia; Fluegge, Daniela; Engelhardt, Corinna H; Lohmer, Stefan; Hatt, Hanns; Corazza, Sabrina; Spehr, Jennifer; Neuhaus, Eva M; Spehr, Marc

2011-01-01

10

Testicular Sertoli cell function in ankylosing spondylitis.  

PubMed

To assess the testicular Sertoli cell function according to inhibin B levels in ankylosing spondylitis (AS) patients and the possible effect of anti-TNF therapy on this hormone production, 20 consecutive AS patients and 24 healthy controls were evaluated. At study entry, AS patients were not receiving sulfasalazine/methotrexate and never have used biological/cytotoxic agents. They were assessed by serum inhibin B levels, hormone profile, urological examination, testicular ultrasound, seminal parameters, and clinical features. Ten of these patients received anti-TNF treatment and they were reevaluated for Sertoli function and disease parameters at 6 months. Four of them agreed to repeat sperm analysis. At study entry, the median of inhibin B (68 vs. 112.9 pg/mL, p = 0.111), follicle-stimulating hormone levels (3.45 vs. 3.65 IU/L, p = 0.795), and the other hormones was comparable in AS patients and controls (p > 0.05). Sperm analysis was similar in AS patients and controls (p > 0.05) with one AS patient presenting borderline low inhibin B levels. Further analysis at 6 months of the 10 patients referred for anti-TNF therapy, including one with borderline inhibin B, revealed that median inhibin B levels remained stable (116.5 vs. 126.5 pg/mL, p = 0.431) with a significant improvement in C-reactive protein (27.8 vs. 2.27 mg/L, p = 0.039). Sperm motility and concentration were preserved in the four patients who repeated this analysis after TNF blockage. In conclusion, this was the first study to report, using a specific marker, a normal testicular Sertoli cell function in AS patients with mild to moderate disease activity. PMID:23417428

Almeida, Breno Pires; Saad, Carla Gonçalves Schahin; Souza, Fernando Henrique Carlos; Moraes, Julio Cesar Bertacini; Nukumizu, Lucia Akemi; Viana, Vilma Santos Trindade; Bonfá, Eloísa; Silva, Clovis Artur

2013-07-01

11

Understanding the role of thyroid hormone in Sertoli cell development: a mechanistic hypothesis  

Microsoft Academic Search

More than a decade of research has shown that Sertoli cell proliferation is regulated by thyroid hormone. Neonatal hypothyroidism lengthens the period of Sertoli cell proliferation, leading to increases in Sertoli cell number, testis weight, and daily sperm production (DSP) when euthyroidism is re-established. In contrast, the neonatal Sertoli cell proliferative period is shortened under hyperthyroid conditions, but the mechanism

Denise R. Holsberger; Paul S. Cooke

2005-01-01

12

In vivo actions of the Sertoli cell glucocorticoid receptor.  

PubMed

We determined the functional role of the Sertoli cell glucocorticoid receptor (GR) in vivo using a transgenic Cre-loxP approach to conditionally disrupt GR expression. Sertoli cell GR knockout (SCGRKO) was shown by absent Sertoli cell-specific GR immunolocalization and reduced levels of glucocorticoid-responsive Stc1 and Tsc22d3 mRNA in SCGRKO relative to control testes. Adult SCGRKO testes exhibited distinct morphological changes, including reduced seminiferous tubular lumen formation, decreased total Sertoli cell numbers, and parallel reductions in meiotic spermatocyte and postmeiotic spermatid numbers. Conversely, tubular diameter was increased and testis size was normal in SCGRKO males. Decreased serum FSH and testicular Fshr mRNA levels were consistent with reduced Sertoli cell number. Adult SCGRKO testes also displayed atypical germ cells and interstitial focal accumulations of hypertrophic lipid-laden, immature-like Leydig cells. Circulating LH, and testicular Lhr mRNA, testosterone, dihydrotestosterone, and 3?/3?-diol levels were all reduced in mature SCGRKO mice, whereas serum testosterone and dihydrotestosterone levels remained normal. Moreover, Sertoli cell GR disruption caused differential changes to steroidogenic enzyme transcripts, with down-regulated testicular Cyp11a1 contrasting with up-regulated Hsd17b3 expression. Reduced SCGRKO testicular expression of Hsd11b2, encoding an enzyme for corticosterone inactivation, supports a dynamic coupling between Hsd11b and androgen production. Our novel SCGRKO model has revealed that Sertoli cell-mediated GR actions support normal testicular function. Sertoli cell GR is required to maintain normal testicular Sertoli/germ cell numbers and circulating gonadotropin levels, as well as optimal Leydig cell maturation and steroidogenesis, providing new insight into gluocorticoid-mediated impact on male reproduction. PMID:24424066

Hazra, Rasmani; Upton, Dannielle; Jimenez, Mark; Desai, Reena; Handelsman, David J; Allan, Charles M

2014-03-01

13

Characterization and functionality of proliferative human Sertoli cells.  

PubMed

It has long been thought that mammalian Sertoli cells are terminally differentiated and nondividing postpuberty. For most previous in vitro studies immature rodent testes have been the source of Sertoli cells and these have shown little proliferative ability when cultured. We have isolated and characterized Sertoli cells from human cadaveric testes from seven donors ranging from 12 to 36 years of age. The cells proliferated readily in vitro under the optimized conditions used with a doubling time of approximately 4 days. Nuclear 5-ethynyl-2'-deoxyuridine (EdU) incorporation confirmed that dividing cells represented the majority of the population. Classical Sertoli cell ultrastructural features, lipid droplet accumulation, and immunoexpression of GATA-4, Sox9, and the FSH receptor (FSHr) were observed by electron and fluorescence microscopy, respectively. Flow cytometry revealed the expression of GATA-4 and Sox9 by more than 99% of the cells, and abundant expression of a number of markers indicative of multipotent mesenchymal cells. Low detection of endogenous alkaline phosphatase activity after passaging showed that few peritubular myoid cells were present. GATA-4 and SOX9 expression were confirmed by reverse transcription polymerase chain reaction (RT-PCR), along with expression of stem cell factor (SCF), glial cell line-derived neurotrophic factor (GDNF), and bone morphogenic protein 4 (BMP4). Tight junctions were formed by Sertoli cells plated on transwell inserts coated with fibronectin as revealed by increased transepithelial electrical resistance (TER) and polarized secretion of the immunoregulatory protein, galectin-1. These primary Sertoli cell populations could be expanded dramatically in vitro and could be cryopreserved. The results show that functional human Sertoli cells can be propagated in vitro from testicular cells isolated from adult testis. The proliferative human Sertoli cells should have important applications in studying infertility, reproductive toxicology, testicular cancer, and spermatogenesis, and due to their unique biological properties potentially could be useful in cell therapy. PMID:21054948

Chui, Kitty; Trivedi, Alpa; Cheng, C Yan; Cherbavaz, Diana B; Dazin, Paul F; Huynh, Ai Lam Thu; Mitchell, James B; Rabinovich, Gabriel A; Noble-Haeusslein, Linda J; John, Constance M

2011-01-01

14

Characterization and Functionality of Proliferative Human Sertoli Cells  

PubMed Central

It has long been thought that mammalian Sertoli cells are terminally differentiated and nondividing postpuberty. For most previous in vitro studies immature rodent testes have been the source of Sertoli cells and these have shown little proliferative ability when cultured. We have isolated and characterized Sertoli cells from human cadaveric testes from seven donors ranging from 12 to 36 years of age. The cells proliferated readily in vitro under the optimized conditions used with a doubling time of approximately 4 days. Nuclear 5-ethynyl-2?-deoxyuridine (EdU) incorporation confirmed that dividing cells represented the majority of the population. Classical Sertoli cell ultrastructural features, lipid droplet accumulation, and immunoexpression of GATA-4, Sox9, and the FSH receptor (FSHr) were observed by electron and fluorescence microscopy, respectively. Flow cytometry revealed the expression of GATA-4 and Sox9 by more than 99% of the cells, and abundant expression of a number of markers indicative of multipotent mesenchymal cells. Low detection of endogenous alkaline phosphatase activity after passaging showed that few peritubular myoid cells were present. GATA-4 and SOX9 expression were confirmed by reverse transcription polymerase chain reaction (RT-PCR), along with expression of stem cell factor (SCF), glial cell line-derived neurotrophic factor (GDNF), and bone morphogenic protein 4 (BMP4). Tight junctions were formed by Sertoli cells plated on transwell inserts coated with fibronectin as revealed by increased transepithelial electrical resistance (TER) and polarized secretion of the immunoregulatory protein, galectin-1. These primary Sertoli cell populations could be expanded dramatically in vitro and could be cryopreserved. The results show that functional human Sertoli cells can be propagated in vitro from testicular cells isolated from adult testis. The proliferative human Sertoli cells should have important applications in studying infertility, reproductive toxicology, testicular cancer, and spermatogenesis, and due to their unique biological properties potentially could be useful in cell therapy.

Chui, Kitty; Trivedi, Alpa; Cheng, C. Yan; Cherbavaz, Diana B.; Dazin, Paul F.; Huynh, Ai Lam Thu; Mitchell, James B.; Rabinovich, Gabriel A.; Noble-Haeusslein, Linda J.; John, Constance M.

2014-01-01

15

GATA4 regulates Sertoli cell function and fertility in adult male mice  

PubMed Central

Transcription factor GATA4 is expressed in Sertoli and Leydig cells and is required for proper development of the murine fetal testis. The role of GATA4 in adult testicular function, however, has remained unclear due to prenatal lethality of mice harboring homozygous mutations in Gata4. To characterize the function of GATA4 in the adult testis, we generated mice in which Gata4 was conditionally deleted in Sertoli cells using Cre-LoxP recombination with Amhr2-Cre. Conditional knockout (cKO) mice developed age-dependent testicular atrophy and loss of fertility, which coincided with decreases in the quantity and motility of sperm. Histological analysis demonstrated Sertoli cell vacuolation, impaired spermatogenesis, and increased permeability of the blood-testis barrier. RT-PCR analysis of cKO testes showed decreased expression of germ cell markers and increased expression of testicular injury markers. Our findings support the premise that GATA4 is a key transcriptional regulator of Sertoli cell function in adult mice.

Kyronlahti, Antti; Euler, Rosemarie; Bielinska, Malgorzata; Schoeller, Erica L.; Moley, Kelle H.; Toppari, Jorma; Heikinheimo, Markku; Wilson, David B.

2011-01-01

16

Extracellular matrix regulates Sertoli cell differentiation, testicular cord formation, and germ cell development in vitro  

PubMed Central

Sertoli cell preparations isolated from 10-day-old rats were cultured on three different substrates: plastic, a matrix deposited by co- culture of Sertoli and peritubular myoid cells, and a reconstituted basement membrane gel from the EHS tumor. When grown on plastic, Sertoli cells formed a squamous monolayer that did not retain contaminating germ cells. Grown on the matrix deposited by Sertoli- myoid cell co-cultures, Sertoli cells were more cuboidal and supported some germ cells but did not allow them to differentiate. After 3 wk however, the Sertoli cells flattened to resemble those grown on plastic. In contrast, the Sertoli cells grown on top of the reconstituted basement membrane formed polarized monolayers virtually identical to Sertoli cells in vivo. They were columnar with an elaborate cytoskeleton. In addition, they had characteristic basally located tight junctions and maintained germ cells for at least 5 wk in the basal aspect of the monolayer. However, germ cells did not differentiate. Total protein, androgen binding protein, transferrin, and type I collagen secretion were markedly greater when Sertoli cells were grown on the extracellular matrices than when they were grown on plastic. When Sertoli cells were cultured within rather than on top of reconstituted basement membrane gels they reorganized into cords. After one week, tight junctional complexes formed between adjacent Sertoli cells, functionally compartmentalizing the cords into central (adluminal) and peripheral (basal) compartments. Germ cells within the cords continued to differentiate. Thus, Sertoli cells cultured on top of extracellular matrix components assume a phenotype and morphology more characteristic of the in vivo, differentiated cells. Growing Sertoli cells within reconstituted basement membrane gels induces a morphogenesis of the cells into cords, which closely resemble the organ from which the cells were dissociated and which provide an environment permissive for germ cell differentiation.

1985-01-01

17

Expression of the Pem Homeobox Gene in Sertoli Cells Increases the Frequency of Adjacent Germ Cells with Deoxyribonucleic Acid Strand Breaks  

Microsoft Academic Search

Pem is a member of the homeobox transcription factor family that is expressed in somatic cells in male and female repro- ductive tissues. In the murine testis, Pem is specifically ex- pressed in Sertoli cells, where it is dramatically induced at the initiation of meiosis during the first wave of spermatogenesis and then later is restricted to stages IV-VIII of

CHAD M. WAYNE; KEITH SUTTON; MILES F. WILKINSON

2010-01-01

18

Lentiviral transduction of rat Sertoli cells as a means to modify gene expression.  

PubMed

Primary cell culture is an established and widely used technique to study Sertoli cell function in vitro. However, the relative difficulty of stably overexpressing or knocking down genes in Sertoli cell culture has limited progress in the field. In this technical report, we present a method to transduce 20 dpp rat Sertoli cell cultures with VSV-G pseudotyped lentiviral based vectors at a high rate (~80%), with stable reporter gene expression. Although high transgene expression is desirable, it was noted that at transduction rates > 60% inter-Sertoli cell tight junction integrity and, hence, Sertoli cell function, were transiently compromised. We envisage that this optimized procedure has the potential to stimulate Sertoli cell research, and motivate the use of Sertoli cells in various cell therapy applications. PMID:23248769

Nicholls, Peter K; Stanton, Peter G; Rainczuk, Katarzyna E; Qian, Hongwei; Gregorevic, Paul; Harrison, Craig A

2012-10-01

19

Gene silencing by RNAi in mouse Sertoli cells  

PubMed Central

Background RNA interference (RNAi) is a valuable tool in the investigation of gene function. The purpose of this study was to examine the availability, target cell types and efficiency of RNAi in the mouse seminiferous epithelium. Methods The experimental model was based on transgenic mice expressing EGFP (enhanced green fluorescent protein). RNAi was induced by in vivo transfection of plasmid vectors encoding for short hairpin RNAs (shRNAs) targeting EGFP. shRNAs were transfected in vivo by microinjection into the seminiferous tubules via the rete testis followed by square wave electroporation. As a transfection reporter, expression of red fluorescent protein (HcRed 1) was used. Cell types, the efficiency of both transfections and RNAi were all evaluated. Results Sertoli cells were the main transfected cells. A reduction of about 40% in the level of EGFP protein was detected in cells successfully transfected both in vivo and in vitro. However, the efficiency of in vivo transfection was low. Conclusion In adult seminiferous epithelial cells, in vivo post-transcriptional gene silencing mediated by RNAi via shRNA is efficient in Sertoli cells. Similar levels of RNAi were detected both in vivo and in vitro. This also indicates that Sertoli cells have the necessary silencing machinery to repress the expression of endogenous genes via RNAi.

Gonzalez-Gonzalez, Emilio; Lopez-Casas, Pedro P; del Mazo, Jesus

2008-01-01

20

Granular transformation of Sertoli cells in testicular disorders.  

PubMed

In order to study the granular transformation of Sertoli cells the following testicular specimens were reviewed: 58 postmortem biopsies from 21 children and 37 young adult males with normal histologic pattern; 165 biopsies from prepubertal cryptorchid testes; 38 biopsies and 18 surgical specimens from postpubertal-cryptorchid testes; bilateral biopsies from eight men with Del Castillo's syndrome, 14 men with retractile testes, and five men with obstructive azospermia; 17 bilateral and seven unilateral biopsies from 24 men with varicocele; seven unilateral biopsies plus five surgical specimens from 12 men with male pseudohermaphroditism; one biopsy and one surgical specimen from two men with macroorchidism; and the autopsy specimens from 28 adult men with acquired immunodeficiency syndrome (AIDS). Sertoli cells with eosinophilic granular cytoplasm were found in the testes of one prepubertal and four postpubertal cryptorchid males, two males with Del Castillo's syndrome, two males with retractile testes, four males with varicocele, two male pseudohermaphrodites, two males with macroorchidism, and one male with AIDS and interstitial orchitis. Histochemical and ultrastructural examination of granular Sertoli cells revealed that these cells accumulate secondary lysosomes and show scant cytoplasmic organelles. In the males with varicocele or retractile testes, these lysosomes were probably heterolysosomes that had degraded the germ cells and testicular fluid accumulated in the lumen of the ectatic seminiferous tubules of these testes. A similar mechanism is also probable in the male with interstitial orchitis that had caused germ cell destruction. In the other cases, in which the tubules showed reduced lumen and severe germ cell depletion, the abundant lysosomes are probably cytolysosomes. The development of these cytolysosomes might be related to the Sertoli cell dysgenesis present in these testes. PMID:1672120

Nistal, M; Garcia-Rodeja, E; Paniagua, R

1991-02-01

21

Autoimmune orchitis can be induced by thymic lymphocytes autosensitized against syngeneic sertoli cells in vitro.  

PubMed

Thymic lymphocytes from normal inbred Lewis/Wistar rats were cocultured with syngeneic Sertoli cell-peritubular cell preparations in the presence of heterologous or allogeneic serum. Thymic cells cultured in this manner bound to Sertoli cells, became autosensitized , and markedly altered syngeneic Sertoli cell surface properties and remodeling functions in vitro. In contrast, control thymic cells incubated with Sertoli cells in autologous or syngeneic serum did not become sensitized. Coculture of autosensitized thymic cells with syngeneic seminiferous tubule segments, or local transfer of such lymphocytes into syngeneic rat testes, resulted in intratubular infiltration by "light cells." Intratesticular injection of autosensitized thymic cells was followed by derangement of the seminiferous epithelium, and by morphologic changes characteristic of experimental autoimmune orchitis. Thymic cells incubated with Sertoli cells in autologous or syngeneic serum did not elicit these changes. Thymic cells incubated with peritubular cells in heterologous or autologous serum behaved like control thymocytes, and were not sensitized. Data presented indicate that thymic cells are potentially capable of recognizing syngeneic Sertoli cell self-antigens. We speculate that factors normally present in serum may inhibit the recognition by thymic lymphocytes of antigenic determinants present on Sertoli cells. We discuss the possibility that the modulation of interactions between immature thymic lymphocytes and Sertoli cells is implicated in the prevention of autoimmune reactions against the testis. PMID:6203564

Tung, P S; Fritz, I B

1984-05-01

22

Polycyclic Aromatic Hydrocarbon-Induced Cytotoxicity in Cultured Rat Sertoli Cells Involves Differential Apoptotic Response  

Microsoft Academic Search

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous and persistent environmental contami- nants. Some PAHs are carcinogens and may affect the male reproductive system. Therefore, we exposed cultured rat Sertoli cells to a variety of PAHs to determine possible direct toxic effects on the cells of the seminiferous epithelium. Sertoli cells were chosen because they support germ cell development and maintain spermatogenesis.

Samir S. Raychoudhury; Dana Kubinski

2002-01-01

23

Effects of simulated microgravity on mouse Sertoli cells in culture  

NASA Astrophysics Data System (ADS)

With the advent of space flights questions concerning the effects of microgravity (0xG) on hu-man reproduction physiology have got priority Spermatogenesis is a complex, highly ordered process of cell division and differentiation by which spermatogonial cells give rise to mature spermatozoa. Sertoli cells play a crucial role in the development of germ cells and the regulation of spermatogenesis. In this study the influence of 0xG on Sertoli cells was evaluated. A Sertoli cell line from mouse testis (42GPA9) was analyzed for cytoskeletal (using the 3D reconstruction generated from a stack of confocal images) and SHBG changes by immunohistochemistry, for antioxidant agents by RT-PCR and for culture medium lactate concentrations by wet chemistry. Cells were cultured for 6, 24 and 48 hrs on a three-dimensional Random Positioning Machine (3D-RPM); static controls (1xG) were positioned on the supporting frame. At the end of each experiment, cultured cells were either fixed in paraformaldehyde or RNA-extracted or used for culture medium lactate measurements as needed. At 0xG Sertoli cytoskeleton got disorganized, microtubules fragmented and SHBG undetectable already after 24 hrs, with alterations wors-ening further until 48 hrs; various antioxidant systems (SOD, GST, PARP, MTs) appreciably increased during the first 24 hrs but significantly decreased at 48 hrs. No changes occurred in 1xG samples. At least initially, 0xG seems to perturb antioxidant protection strategies allowing the testes to support sperm production, thus generating an aging-like state of oxidative stress. Lactate production at 0xG slightly decreased only after 24 hrs. Further experiments need to be carried out in space to investigate upon steroidogenesis and germ cell differentiation within the testis, to rule out eventually pending male infertility consequences, which would be a problem nowadays, when life expectancy increases and male fertility might become a social issue often extending into 60 years and over. (experiment funded by ASI, through a grant within the OSMA project).

Angela, Masini Maria; Prato, Paola; Linda, Scarabelli; Lanza, Cristina; Palmero, Silvio; Pointis, Georges; Ricci, Franco; Strollo, Felice

24

Regulation by Thyroid Hormone of the Expression of Basement Membrane Components in Rat Prepubertal Sertoli Cells  

Microsoft Academic Search

The present study reports the modulation of basement membrane (BM) components, laminin, entactin, and type IV collagen, expression in prepubertal rat Sertoli cell by the thyroid hormone T3. Immuno- cytochemical studies of permeabilized Sertoli cells in culture showed that T3 treatment (10 27 M for 24 h) increased the number of cells staining positive for laminin and\\/or entactin (from 58

SALVATORE ULISSE; NADIA RUCCI; ELEONORA CAROSA; FILOMENA M. GRAZIANO; ANTONIO PAVAN; PIERGIUSEPPE CEDDIA; MARIO ARIZZI; PAOLA MUZI; LUISA CIRONI; LUCIO GNESSI; EMMANUELE A. JANNINI

1998-01-01

25

In vitro effects of simulated microgravity on Sertoli cell function  

NASA Astrophysics Data System (ADS)

With the advent of space flights questions concerning the effects of microgravity (0×G) on human reproductive physiology have received great attention. The aim of this study was to evaluate the influence of 0×G on Sertoli cells. A Sertoli cell line from mouse testis (42GPA9) was analyzed for cytoskeletal and Sex Hormone Binding Globilin (SHBG) changes by immunohistochemistry, for antioxidant content by RT-PCR and for culture medium lactate concentrations by protein chemistry. Cells were cultured for 6, 24 and 48 h on a three-dimensional Random Positioning Machine (3D-RPM); static controls (1×G) were positioned on the supporting frame. At the end of each experiment, cultured cells were either fixed in paraformaldehyde or lysed and RNA-extracted or used for culture medium lactate measurements as needed. At 0×G, Sertoli cytoskeleton became disorganized, microtubules fragmented and SHBG undetectable already after 24 h, with alterations worsening by 48 h. It was evident that various antioxidant systems appreciably increased during the first 24 h but significantly decreased at 48 h. No changes occurred in the 1×G samples. Initially, 0×G seemed to disturb antioxidant protection strategies allowing the testes to support sperm production, thus generating an aging-like state of oxidative stress. Lactate production at 0×G slightly decreased after 24 h. Further experiments are needed in space to investigate upon steroidogenesis and germ cell differentiation within the testis, to rule out male infertility as a possible consequence, which could be a problem, as life expectancy increases.

Masini, M. A.; Prato, P.; Scarabelli, L.; Lanza, C.; Palmero, S.; Pointis, G.; Ricci, F.; Strollo, F.

2011-02-01

26

Sertoli-Sertoli and Sertoli-germ cell interactions and their significance in germ cell movement in the seminiferous epithelium during spermatogenesis.  

PubMed

Spermatogenesis is the process by which a single spermatogonium develops into 256 spermatozoa, one of which will fertilize the ovum. Since the 1950s when the stages of the epithelial cycle were first described, reproductive biologists have been in pursuit of one question: How can a spermatogonium traverse the epithelium, while at the same time differentiating into elongate spermatids that remain attached to the Sertoli cell throughout their development? Although it was generally agreed upon that junction restructuring was involved, at that time the types of junctions present in the testis were not even discerned. Today, it is known that tight, anchoring, and gap junctions are found in the testis. The testis also has two unique anchoring junction types, the ectoplasmic specialization and tubulobulbar complex. However, attention has recently shifted on identifying the regulatory molecules that "open" and "close" junctions, because this information will be useful in elucidating the mechanism of germ cell movement. For instance, cytokines have been shown to induce Sertoli cell tight junction disassembly by shutting down the production of tight junction proteins. Other factors such as proteases, protease inhibitors, GTPases, kinases, and phosphatases also come into play. In this review, we focus on this cellular phenomenon, recapping recent developments in the field. PMID:15466940

Mruk, Dolores D; Cheng, C Yan

2004-10-01

27

Direct reprogramming of fibroblasts into embryonic Sertoli-like cells by defined factors  

PubMed Central

SUMMARY Sertoli cells are considered the “supporting cells” of the testis that play an essential role in sex determination during embryogenesis and in spermatogenesis during adulthood. Their essential roles in male fertility along with their immunosuppressive and neurotrophic properties make them an attractive cell type for therapeutic applications. Here we demonstrate the generation of embryonic sertoli-like cells (ieSCs) by ectopic expression of five transcription factors. We characterize the role of specific transcription factor combinations in the transition from fibroblasts to ieSCs and identify key steps in the process. Initially, transduced fibroblasts underwent a mesenchymal to epithelial transition and then, acquired the ability to aggregate, formed tubular-like structures, and expressed embryonic Sertoli specific markers. These Sertoli-like cells facilitated neuronal differentiation and self-renewal of NPC, supported the survival of germ cells in culture and cooperated with endogenous embryonic Sertoli and primordial germ cells in the generation of testicular cords in the fetal gonad.

Buganim, Yosef; Itskovich, Elena; Hu, Yueh-Chiang; Cheng, Albert W.; Ganz, Kibibi; Sarkar, Sovan; Fu, Dongdong; Welstead, Grant; Page, David C.; Jaenisch, Rudolf

2012-01-01

28

A survey of Sertoli cell differentiation in men after gonadotropin suppression and in testicular cancer  

PubMed Central

It is widely held that the somatic cell population that is responsible for sperm development and output (Sertoli cells) is terminally differentiated and unmodifiable in adults. It is postulated, with little evidence, that Sertoli cells are not terminally differentiated in some phenotypes of infertility and testicular cancer. This study sought to compare markers of Sertoli cell differentiation in normospermic men, oligospermic men (undergoing gonadotropin suppression) and testicular carcinoma in situ (CIS) and seminoma samples. Confocal microscopy was used to assess the expression of markers of proliferation (PCNA and Ki67) and functional differentiation (androgen receptor). As additional markers of differentiation, the organization of Sertoli cell tight junction and associated proteins were assessed in specimens with carcinoma in situ. In normal men, Sertoli cells exhibited a differentiated phenotype (i.e., PCNA and Ki67 negative, androgen 40 receptor positive). However, after long-term gonadotropin suppression, 1.7 ± 0.6% of Sertoli cells exhibited PCNA reactivity associated with a diminished immunoreactivity in androgen receptor, suggesting an undifferentiated phenotype. Ki67-positive Sertoli cells were also observed. PCNA-positive Sertoli cells were never observed in tubules with carcinoma in situ, and only rarely observed adjacent to seminoma. Tight junction protein localization (claudin 11, JAM-A and ZO-1) was altered in CIS, with a reduction in JAM-A reactivity in Sertoli cells from tubules with CIS and the emergence of strong JAM-A reactivity in seminoma. These findings indicate that adult human Sertoli cells exhibit characteristics of an undifferentiated state in oligospermic men and patients with CIS and seminoma in the presence of germ cell neoplasia.

Tarulli, Gerard A.; Stanton, Peter G.; Loveland, Kate L.; Meyts, Ewa Rajpert-De; McLachlan, Robert I.; Meachem, Sarah J.

2013-01-01

29

A survey of Sertoli cell differentiation in men after gonadotropin suppression and in testicular cancer.  

PubMed

It is widely held that the somatic cell population that is responsible for sperm development and output (Sertoli cells) is terminally differentiated and unmodifiable in adults. It is postulated, with little evidence, that Sertoli cells are not terminally differentiated in some phenotypes of infertility and testicular cancer. This study sought to compare markers of Sertoli cell differentiation in normospermic men, oligospermic men (undergoing gonadotropin suppression) and testicular carcinoma in situ (CIS) and seminoma samples. Confocal microscopy was used to assess the expression of markers of proliferation (PCNA and Ki67) and functional differentiation (androgen receptor). As additional markers of differentiation, the organization of Sertoli cell tight junction and associated proteins were assessed in specimens with carcinoma in situ. In normal men, Sertoli cells exhibited a differentiated phenotype (i.e., PCNA and Ki67 negative, androgen 40 receptor positive). However, after long-term gonadotropin suppression, 1.7 ± 0.6% of Sertoli cells exhibited PCNA reactivity associated with a diminished immunoreactivity in androgen receptor, suggesting an undifferentiated phenotype. Ki67-positive Sertoli cells were also observed. PCNA-positive Sertoli cells were never observed in tubules with carcinoma in situ, and only rarely observed adjacent to seminoma. Tight junction protein localization (claudin 11, JAM-A and ZO-1) was altered in CIS, with a reduction in JAM-A reactivity in Sertoli cells from tubules with CIS and the emergence of strong JAM-A reactivity in seminoma. These findings indicate that adult human Sertoli cells exhibit characteristics of an undifferentiated state in oligospermic men and patients with CIS and seminoma in the presence of germ cell neoplasia. PMID:23687617

Tarulli, Gerard A; Stanton, Peter G; Loveland, Kate L; Meyts, Ewa Rajpert-De; McLachlan, Robert I; Meachem, Sarah J

2013-01-01

30

Intrauterine growth retardation associated with precocious puberty and sertoli cell hyperplasia.  

PubMed

The original description of patients with Russell-Silver syndrome included precocious puberty, the mechanism of which was unclear. We describe a child with a Russell-Silver syndrome-like phenotype who presented with precocious puberty that was associated with hyperplasia of the Sertoli cells. The patient was found to have an immature cryptorchid testicle; hyperplastic Sertoli cells were also aneuploid carrying trisomy 8. This chromosomal abnormality was present in Sertoli cells only and could not be detected in peripheral lymphocytes, tunica vaginalis, or other, normal, testicular tissue. Sertoli cells in culture showed excess aromatization providing an explanation for the rapid advancement of the patient's bone age. We conclude that in a patient with a Russell-Silver syndrome-like phenotype, Sertoli cell hyperplasia was associated with somatic trisomy 8, increased aromatization, and gonadotropin-independent precocious puberty. PMID:20411478

Lodish, M B; Gartner, L A; Albini, P; Sabnis, G; Brodie, A; Meck, J M; Meloni-Ehrig, A M; Hill, S; Tsilou, E; Valera, V A; Walter, B A; Merino, M J; Stratakis, C A

2010-08-01

31

Intrauterine growth retardation associated with precocious puberty and Sertoli cell hyperplasia  

PubMed Central

The original description of patients with Russell-Silver syndrome included precocious puberty, the mechanism of which was unclear. We describe a child with a Russell-Silver syndrome-like phenotype who presented with precocious puberty that was associated with hyperplasia of the Sertoli cells. The patient was found to have an immature cryptorchid testicle; hyperplastic Sertoli cells were also aneuploid carrying trisomy 8. This chromosomal abnormality was present in Sertoli cells only and could not be detected in peripheral lymphocytes, tunica vaginalis, or other, normal, testicular tissue. Sertoli cells in culture showed excess aromatization providing an explanation for the rapid advancement of the patient’s bone age. We conclude that in a patient with a Russell-Silver syndrome-like phenotype, Sertoli cell hyperplasia was associated with somatic trisomy 8, increased aromatization and gonadotropin-independent precocious puberty.

Lodish, Maya B.; Gartner, Lou Ann; Albini, Paul; Brodie, Angela; Meck, Jeanne M.; Meloni-Ehrig, Aurelia M; Hill, Suvimol; Tsilou, Ekaterini; Carney, J. Aidan; Valera, Vladimir A.; Walter, Beatriz A.; Merino, Maria J.; Stratakis, Constantine A.

2012-01-01

32

Heat treatment induces liver receptor homolog-1 expression in monkey and rat sertoli cells.  

PubMed

We demonstrated in this study that liver receptor homolog-1 (LRH-1) was expressed in the round spermatids in normal monkey testis, and no LRH-1 signal was observed in the Sertoli cells. After local warming (43 C) the monkey testis, however, LRH-1 expression was induced in the Sertoli cells in coincidence with activation of cytokeratin 18 (CK-18), a Sertoli cell dedifferentiated marker. Furthermore, we isolated rat primary Sertoli cells from testes at various stages of development and treated with 43 C water in vitro. The changes in LRH-1 as well as CK-18 expression were analyzed by confocal immunohistochemistry and Western blot. The results showed that LRH-1 was stage-dependently expressed in the Sertoli cells; no LRH-1-positive signal was detected in the cells obtained from the testes of adult rat on d 60 after birth when mature spermatozoa in the testis was completed. However, the mature Sertoli cells were warmed at the 43 C water bath for 15 min, and the LRH-1 signal was remarkably induced in a time-dependent manner, just like the changes of CK-18 expression in the Sertoli cells, suggesting that the heat-induced dedifferentiation of the mature Sertoli cells might be related to LRH-1 regulation. LRH-1 expression induced by the heat treatment was completely inhibited by the addition of ERK inhibitor U0126 in the culture, indicating that the heat-induced LRH-1 expression in the Sertoli cells may be regulated via ERK1/2 activation pathway. Testosterone was found to have no such effect on LRH-1 expression in the monkey and rat Sertoli cells. PMID:17170099

Guo, Jian; Tao, Shi-Xin; Chen, Min; Shi, Yu-Qiang; Zhang, Zhu-Qiang; Li, Yin-Chuan; Zhang, Xue-Sen; Hu, Zhao-Yuan; Liu, Yi-Xun

2007-03-01

33

Sertoli Cell Behaviors in Developing Testis Cords and Postnatal Seminiferous Tubules of the Mouse1  

PubMed Central

Sertoli cells are the primary structural component of the fetal testis cords and postnatal seminiferous tubules. Live imaging technologies facilitate the visualization of cell morphologies and behaviors through developmental processes. A transgenic mouse line was generated using a fragment of the rat Gata4 gene to direct the expression of a dual-color fluorescent protein reporter in fetal and adult Sertoli cells. The reporter encoded a red fluorescent protein, monomeric Cherry (mCherry), fused to histone 2B and enhanced green fluorescent protein (EGFP) fused to a glycosylphosphatidylinositol sequence, with a self-cleaving 2A polypeptide separating the two fusion proteins. After translation, the red and green fluorescent proteins translocated to the nucleus and plasma membrane, respectively, of Sertoli cells. Transgene expression in testes was first detected by fluorescent microscopy around Embryonic Day 12.0. Sertoli cell division and migration were visualized during testis cord formation in organ culture. Initially, the Sertoli cells had mesenchyme-like morphologies and behaviors, but later, the cells migrated to the periphery of the testis cords to become epithelialized. In postnatal seminiferous tubules, Sertoli nuclei were evenly spaced when viewed from the external surface of tubules, and Sertoli cytoplasm and membranes were associated with germ cells basally in a rosette pattern. This mouse line was bred to previously described transgenic mouse lines expressing EGFP in Sertoli cytoplasm or a nuclear cyan fluorescent protein (Cerulean) and mCherry in plasma membranes of germ cells. This revealed the physical relationship between Sertoli and germ cells in developing testis cords and provided a novel perspective on Sertoli cell development.

Nel-Themaat, Liesl; Jang, Chuan-Wei; Stewart, M. David; Akiyama, Haruhiko; Viger, Robert S.; Behringer, Richard R.

2010-01-01

34

Sertoli cell behaviors in developing testis cords and postnatal seminiferous tubules of the mouse.  

PubMed

Sertoli cells are the primary structural component of the fetal testis cords and postnatal seminiferous tubules. Live imaging technologies facilitate the visualization of cell morphologies and behaviors through developmental processes. A transgenic mouse line was generated using a fragment of the rat Gata4 gene to direct the expression of a dual-color fluorescent protein reporter in fetal and adult Sertoli cells. The reporter encoded a red fluorescent protein, monomeric Cherry (mCherry), fused to histone 2B and enhanced green fluorescent protein (EGFP) fused to a glycosylphosphatidylinositol sequence, with a self-cleaving 2A polypeptide separating the two fusion proteins. After translation, the red and green fluorescent proteins translocated to the nucleus and plasma membrane, respectively, of Sertoli cells. Transgene expression in testes was first detected by fluorescent microscopy around Embryonic Day 12.0. Sertoli cell division and migration were visualized during testis cord formation in organ culture. Initially, the Sertoli cells had mesenchyme-like morphologies and behaviors, but later, the cells migrated to the periphery of the testis cords to become epithelialized. In postnatal seminiferous tubules, Sertoli nuclei were evenly spaced when viewed from the external surface of tubules, and Sertoli cytoplasm and membranes were associated with germ cells basally in a rosette pattern. This mouse line was bred to previously described transgenic mouse lines expressing EGFP in Sertoli cytoplasm or a nuclear cyan fluorescent protein (Cerulean) and mCherry in plasma membranes of germ cells. This revealed the physical relationship between Sertoli and germ cells in developing testis cords and provided a novel perspective on Sertoli cell development. PMID:20944081

Nel-Themaat, Liesl; Jang, Chuan-Wei; Stewart, M David; Akiyama, Haruhiko; Viger, Robert S; Behringer, Richard R

2011-02-01

35

Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). Black-Right-Pointing-Pointer Presence of SCs dramatically increased proliferation and migration of UCMSCs. Black-Right-Pointing-Pointer Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of 'nurse' cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.

Zhang, Fenxi, E-mail: fxzhang0824@gmail.com [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People's Republic of China (China)] [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People's Republic of China (China); Hong, Yan; Liang, Wenmei [Department of Histology and Embryology, Guiyang Medical University, Guizhou 550004, People's Republic of China (China)] [Department of Histology and Embryology, Guiyang Medical University, Guizhou 550004, People's Republic of China (China); Ren, Tongming [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People's Republic of China (China)] [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People's Republic of China (China); Jing, Suhua [ICU Center, The Third Hospital of Xinxiang Medical University, Henan 453003, People's Republic of China (China)] [ICU Center, The Third Hospital of Xinxiang Medical University, Henan 453003, People's Republic of China (China); Lin, Juntang [Stem Cell Center, Xinxiang Medical University, Henan 453003, People's Republic of China (China)] [Stem Cell Center, Xinxiang Medical University, Henan 453003, People's Republic of China (China)

2012-10-12

36

Highly vectorial secretion of inhibin by primate Sertoli cells in vitro.  

PubMed

Having recently demonstrated highly vectorial and FSH-stimulated inhibin secretion by immature rat Sertoli cells in vitro, we wished to determine if vectorial secretion of inhibin was also characteristic of primate Sertoli cells. By adapting techniques for isolation of Sertoli cells from testes of the immature rat and cynomolgus monkey. Sertoli cells were isolated from immature baboon testes. Sertoli cells were then plated onto matrix-impregnated porous filters and cultured in twin chamber assemblies in fully defined, supplemented HEPES-buffered Eagles medium. Inhibin was measured in conditioned culture media by an heterologous RIA validated for primate inhibins. Throughout 28 days of culture immunoreactive inhibin was readily detectable in the upper chambers whereas inhibin was undetectable or just detectable in the lower chambers. The median ratio of upper to lower chamber inhibin content was 15.3 under basal conditions rising to 41 under FSH stimulation. Inhibin secretion into the upper chamber was increased 2.5 +/- 0.4 times by stimulation with ovine FSH (100 ng/ml). We conclude that immature Sertoli cells from a nonhuman primate demonstrate FSH-responsive and highly vectorial secretion of inhibin almost exclusively into the upper chamber. These data suggest that during maturation mammalian Sertoli cells secrete inhibin vectorially mainly from the apical surface of the cell towards the seminiferous tubular lumen. The predominance of inhibin secretion into the seminiferous tubule during testicular maturation suggests that inhibin may have an important paracrine or autocrine role in the developmental biology of spermiogenesis. PMID:2121770

Handelsman, D J; Spaliviero, J A; Phippard, A F

1990-11-01

37

Intracellular signaling pathways involved in the relaxin-induced proliferation of rat Sertoli cells.  

PubMed

Regulation of Sertoli cell number is a key event to determine normal spermatogenesis. We have previously shown that relaxin and its G-protein coupled receptor RXFP1 are expressed in rat Sertoli cells, and that relaxin stimulates Sertoli cell proliferation. This study examined the mechanisms underlying the mitogenic effect of relaxin in a primary culture of Sertoli cells removed from testes of immature rats. Stimulation with exogenous relaxin increased Sertoli cell number and the expression of the proliferating cell nuclear antigen (PCNA), but did not affect the mRNA level of the differentiation markers cadherins 1 and 2. Relaxin-induced Sertoli cell proliferation was blocked by inhibition of MEK/ERK1/2 or PI3K/AKT pathways, but not by inhibition of PKC or EGFR activity. Relaxin induced a rapid and transient activation of ERK1/2 phosphorylation, which was MEK and SRC-dependent, and involved upstream activation of G(i). AKT activation could be detected 5 min after relaxin stimulation, and was still detected after 24h of stimulation with relaxin. Relaxin-induced AKT phosphorylation was G(i)- but not PKA-dependent, and it was blocked by both PI3K and MEK inhibitors. In conclusion, the mitogenic effect of relaxin in Sertoli cell involves coupling to G(i) and activation of both MEK/ERK1/2 and PI3K/AKT pathways. PMID:22819701

Nascimento, Aline Rosa; Pimenta, Maristela Taliari; Lucas, Thais F G; Royer, Carine; Porto, Catarina Segreti; Lazari, Maria Fatima Magalhaes

2012-09-15

38

Sertoli-Leydig Cell Tumor of Ovary- A Diagnostic Dilemma  

PubMed Central

Sertoli Leydig Cell Tumours (SLCTs) are rare, unilateral, sex cord stromal tumours of ovary, which constitute less than 1% of all the ovarian neoplasms. These tumours can be functionally diverse and they may have heterologous elements. We aim to report a case of a 25-year- old woman who presented with suprapubic pain of 5 days duration, a unilateral adnexal mass, hypertestosteronism without virilization. Intraoperative frozen section of the unilateral salpingo-oophorectomy specimen was suggestive of granulosa cell tumour. Histopathological examination, supplemented with alpha-inhibin immunohistochemistry, was diagnostic of Meyer’s type II SLCT. Clinical presentation, pathology and the diagnostic pitfalls in the present case have been presented with a brief review of literature.

C.S, Rohini Dhanya; Padhi, Somanath; Varghese, Renu G'Boy

2014-01-01

39

Activation of PPAR ? and PPAR ?/? regulates Sertoli cell metabolism.  

PubMed

The purpose of this study was to evaluate the existence of a possible simultaneous regulation of fatty acid (FA) metabolism and lactate production by PPAR ? and PPAR ?/? activation in Sertoli cells (SC). SC cultures obtained from 20-day-old rats were incubated with WY14643 or GW0742-pharmacological activators of PPAR ? and PPAR ?/? respectively. The fatty acid transporter CD36, carnitine palmitoyltransferase 1, long- and medium-chain 3-hydroxyacyl-CoA dehydrogenases mRNA levels were analyzed. An increase in the above-mentioned genes in response to activation of both nuclear receptors was observed. Additionally, PPAR ?/? activation increased lactate production as a consequence of increased pyruvate availability by inhibiting the Pyruvate Dehydrogenase Complex. Altogether, these results suggest that in SC, PPAR ? activation participates in the regulation of FA metabolism. On the other hand, PPAR ?/? activation regulates FA metabolism and lactate production ensuring simultaneously the energetic metabolism for SC and germ cells. PMID:24128860

Regueira, M; Riera, M F; Galardo, M N; Pellizzari, E H; Cigorraga, S B; Meroni, S B

2014-01-25

40

Anchoring device between Sertoli cells and late spermatids in rat seminiferous tubules.  

PubMed

Near the end of spermiogensis, the late spermatids remain attached to the superficial layer of the seminiferous epithelium for an appreciable period of time (i.e., 3 to 4 days). Ths sickle-shaped heads of the spermatids are embedded in an apical process of Sertoli cell cytoplasm which is connected to the rest of the cell by a narrow stalk. In the concavity of the head several long (2-3 mum) and very narrow (50 nm) tubular projections of the spermatid's plasma membrane invaginate the Sertoli cell cytoplasm. These tubular processes terminate by a bulbous swelling which may measure up to 1 mum in diameter. Along the process the plasma membrane of the Sertoli cell is closely apposed to the spermatid's membrane, the intracellular space being only 6-8 nm wide. In the Sertoli cytoplasm immediately surrounding the tubular portion of the structure there is an accumulation of filamentous material, while next to the bulbous extremity there are, at a shrot distance, smooth surfaced cisternae of endoplasmic reticulum. The whole structure was referred to as a tubulobulbar complex. These complexes, of which there are up to 24 per spermatid, appear as these cells complete their migration toward the apex of the Sertoli cells. They disappear just before the release of the spermatids in the lumen of the seminiferous tubule as a result of the fragmentation of the spermatid's plasma membrane followed by a resorption of the Sertoli plasma membrane. Morphological evidence suggests that the Tubulobulbar complexes serve as anchoring devices that retain the spermatids at the surface of the seminiferous epithelium while their dissolution contributes in part to the process of spermiation. Similar tubulobulbar complexes were also formed by the plasma membranes of two adjacent Sertoli cells close to the Sertoli-Sertoli tight junctions near the tubular limiting membrane. PMID:937734

Russell, L; Clermont, Y

1976-07-01

41

Late morfofunctional alterations of the Sertoli cell caused by doxorubicin administered to prepubertal rats  

PubMed Central

Background Doxorubicin is a potent chemotherapeutic drug used against a variety of cancers. It acts through interaction with polymerases and topoisomerase II and free radical production. Doxorubicin activity is not specific to cancer cells and can also damage healthy cells, especially those undergoing rapid proliferation, such as spermatogonia. In previous studies our group showed that etoposide, another topoisomarese II poison, causes irreversible damage to Sertoli cells. Thus, the aim of this study was to address the effects of doxorubicin on Sertoli cell morphology and function and on the seminiferous epithelium cycle when administered to prepubertal rats. Methods Prepubertal rats received the dose of 5?mg/Kg of doxorubicin, which was fractioned in two doses: 3?mg/Kg at 15dpp and 2?mg/Kg at 22dpp. The testes were collected at 40, 64 and 127dpp, fixed in Bouin’s liquid and submitted to transferrin immunolabeling for Sertoli cell function analysis. Sertoli cell morphology and the frequency of the stages of the seminiferous epithelium cycle were analyzed in PAS?+?H-stained sections. Results The rats treated with doxorubicin showed reduction of transferrin labeling in the seminiferous epithelium at 40 and 64dpp, suggesting that Sertoli cell function is altered in these rats. All doxorubicin-treated rats showed sloughing and morphological alterations of Sertoli cells. The frequency of the stages of the seminiferous epithelium cycle was also affected in all doxorubicin-treated rats. Conclusions and discussion These data show that doxorubicin administration during prepuberty causes functional and morphological late damage to Sertoli cells; such damage is secondary to the germ cell primary injury and contributed to enhance the spermatogenic harm caused by this drug. However, additional studies are required to clarify if there is also a direct effect of doxorubicin on Sertoli cells producing a primary damage on these cells.

2012-01-01

42

Sertoli Cell-Specific Deletion of the Androgen Receptor Compromises Testicular Immune Privilege in Mice1  

PubMed Central

In the mammalian testis, meiotic and postmeiotic germ cell antigens are granted immune privilege. Both local immune suppression and specialized intercellular junctions between somatic Sertoli cells have been proposed to contribute to a highly restricted and effective blood-testis barrier (BTB) that helps maintain tolerance to germ cell antigens. Several studies have suggested that androgens play a role in immune suppression, although direct evidence for this is lacking. We previously reported that Sertoli cell-specific ablation of the androgen receptor (Ar) decreases expression of Cldn3, an androgen-regulated gene and component of Sertoli cell tight junctions, and increases the permeability of the BTB to biotin, a small-molecular-weight tracer. The physiological consequences of Sertoli cell-specific Ar (S-Ar) ablation on immune privilege are unknown. Here we show that in the testes of S-Ar mutant mice, the ultrastructure of Sertoli cell tight junctions is defective and testicular IgG levels are elevated. The interstitium of S-Ar mutant testes becomes populated with macrophages, neutrophils, plasma cells, and eosinophils, and serum samples of mutant mice contain antibodies against germ cell antigens. Together, these results suggest that Sertoli cell-specific deletion of the androgen receptor results in loss of testicular immune privilege. Suppressed levels of androgen signaling may be a contributing factor in idiopathic male infertility.

Meng, Jing; Greenlee, Anne R.; Taub, Chloe J.; Braun, Robert E.

2011-01-01

43

Krüppel-like factor 4 is involved in functional differentiation of testicular Sertoli cells.  

PubMed

Krüppel-like factor 4 (KLF4) is a pleiotropic zinc finger transcription factor that regulates genes being involved in differentiation and cell-cycle control. Knockout studies revealed a critical function for KLF4 in the terminal differentiation of many epithelial cells. In testicular Sertoli cells, Klf4 is strongly inducible by the glycoprotein follicle stimulating hormone (FSH). Because KLF4 is essential for postnatal survival in mice, we deleted Klf4 specifically in Sertoli cells using the Cre/loxP system. Importantly, around postnatal day 18, a critical period of terminal Sertoli cell differentiation, mutant seminiferous tubules exhibited a disorganized germinal epithelium and delayed lumen formation. The ultrastructural finding of highly vacuolized Sertoli cell cytoplasm and the identification of differentially expressed genes, which are known to play roles during vesicle transport and fusion or for maintenance of the differentiated cell state, suggest impaired apical secretion of the Sertoli cell. Interestingly, a high proportion of all identified genes was localized in a small subregion of chromosome 7, suggesting coordinated regulation. Intriguingly, adult mutant mice are fertile and show normal testicular morphology, although the testosterone levels are decreased. In summary, KLF4 plays a significant role for proper and timely Sertoli cell differentiation in pubertal mice. PMID:18243172

Godmann, Maren; Katz, Jonathan P; Guillou, Florian; Simoni, Manuela; Kaestner, Klaus H; Behr, Rüdiger

2008-03-15

44

RAB13 regulates Sertoli cell permeability barrier dynamics through protein kinase A.  

PubMed

In mammalian testes, the blood-testis barrier (BTB), created by specialized junctions between Sertoli cells near the basement membrane of the seminiferous epithelium, provides an indispensable immune-privileged microenvironment for spermatid development. However, the BTB must experience restructuring during the epithelial cycle to facilitate the transit of preleptotene spermatocytes upon the testosterone-induced new TJ fibrils forming behind these cells, which is intimately related to the extensive dynamics of junction protein complexes between Sertoli cells. As key regulators of protein traffic, Rab GTPases participate in delivery of proteins between distinct cellular sites and cross talk with proteins that constitute tight junction and adherens junction. Using primarily cultured Sertoli cells in vitro with an established tight junction permeability barrier that mimics the BTB in vivo, RAB13 was shown to decrease during the testosterone-induced TJ integrity enhancement, accompanied with an increment in protein kinase A (PKA) activity. Furthermore, knockdown of Rab13 was found to resemble the effect of testosterone on Sertoli cell TJ permeability by reinforcing filamentous actin and occludin distribution at the cell-cell interface and promoting the direct interaction between ZO-1 and occludin. Interestingly, the effects of testosterone and Rab13 knockdown on Sertoli cell epithelium were revealed to be antagonized by PKA activity inhibition. In summary, RAB13 serves as a regulatory component in the assembly and restructuring of the TJ fibrils between adjacent Sertoli cells. PMID:23419316

Su, Wenhui; Liu, Xinchun

2013-06-01

45

Constitutive activation of NOTCH1 signaling in Sertoli cells causes gonocyte exit from quiescence  

PubMed Central

Notch signaling components have long been detected in Sertoli and germ cells in the developing and mature testis. However, the role of this pathway in testis development and spermatogenesis remains unknown. Using reporter mice expressing green fluorescent protein following Notch receptor activation, we found that Notch signaling was active in Sertoli cells at various fetal, neonatal, and adult stages. Since Notch signaling specifies stem cell fate in many developing and mature organ systems, we hypothesized that maintenance and differentiation of gonocytes and/or spermatogonial stem cells would be modulated through this pathway in Sertoli cells. To this end, we generated mutant mice constitutively expressing the active, intracellular domain of NOTCH1 (NICD1) in Sertoli cells. We found that mutant Sertoli cells were morphologically normal before and after birth, but presented a number of functional changes that drastically affected gonocyte numbers and physiology. We observed aberrant exit of gonocytes from mitotic arrest, migration toward cord periphery, and premature differentiation before birth. These events, presumably unsupported by the cellular microenvironment, were followed by gonocyte apoptosis and near complete disappearance of the gonocytes by day 2 after birth. Molecular analysis demonstrated that these effects are correlated with a dysregulation of Sertoli-expressed genes that are required for germ cell maintenance, such as Cyp26b1 and Gdnf. Taken together, our results demonstrate that Notch signaling is active in Sertoli cells throughout development and that proper regulation of Notch signaling in Sertoli cells is required for the maintenance of gonocytes in an undifferentiated state during fetal development.

Garcia, Thomas Xavier; DeFalco, Tony; Capel, Blanche; Hofmann, Marie-Claude

2013-01-01

46

The microtubule plus end-binding protein EB1 is involved in Sertoli cell plasticity in testicular seminiferous tubules.  

PubMed

Sertoli cells of testis belong to a unique type of polarized epithelial cells and are essential for spermatogenesis. They form the blood-testis barrier at the base of seminiferous tubule. Their numerous long, microtubule-rich processes extend inward and associate with developing germ cells to sustain germ cell growth and differentiation. How Sertoli cells develop and maintain their elaborate processes has been an intriguing question. Here we showed that, by microinjecting lentiviral preparations into mouse testes of 29 days postpartum, we were able to specifically label individual Sertoli cells with GFP, thus achieving a clear view of their natural configurations together with associated germ cells in situ. Moreover, compared to other microtubule plus end-tracking proteins such as CLIP-170 and p150(Glued), EB1 was highly expressed in Sertoli cells and located along microtubule bundles in Sertoli cell processes. Stable overexpression of a GFP-tagged dominant-negative EB1 mutant disrupted microtubule organizations in cultured Sertoli cells. Furthermore, its overexpression in testis Sertoli cells altered their shapes. Sertoli cells in situ became rod-like, with decreased basal and lateral cell processes. Seminiferous tubule circularity and germ cell number were also reduced. These data indicate a requirement of proper microtubule arrays for Sertoli cell plasticity and function in testis. PMID:17964570

Wang, Fubin; Zhang, Qiangge; Cao, Jingli; Huang, Qiongping; Zhu, Xueliang

2008-01-01

47

Co-culture of Mouse Embryonic Stem Cells with Sertoli Cells Promote in vitro Generation of Germ Cells  

PubMed Central

Objective(s): Sertoli cells support in vivo germ cell production; but, its exact mechanism has not been well understood. The present study was designed to analyze the effect of Sertoli cells in differentiation of mouse embryonic stem cells (mESCs) to germ cells. Materials and Methods: A fusion construct composed of a Stra8 gene promoter and the coding region of enhanced green fluorescence protein was produced to select differentiated mESCs. To analyze sertoli cells’ effect in differentiation process, mESCs were separated into two groups: the first group was cultured on gelatin with retinoic acid treatment and the second group was co-cultured with sertoli cell feeder without retinoic acid induction. Expressions of pre-meiotic (Stra8), meiotic (Dazl and Sycp3) and post-meiotic (Prm1) genes were evaluated at different differentiation stages (+7, +12 and +18 days of culture). Results: In the first group, expressions of meiotic and post-meiotic genes started 12 and 18 days after induction with retinoic acid, respectively. In the second group, 7 days after co-culturing with Sertoli cells, expression of meiotic and post-meiotic genes was observed. Conclusion: These results show that differentiation process to germ cells is supported by Sertoli cells. Our findings provide a novel effective approach for generation of germ cell in vitro and studying the interaction of germ cells with their niche.

Miryounesi, Mohammad; Nayernia, Karim; Dianatpour, Mahdi; Mansouri, Fatemeh; Modarressi, Mohammad Hossein

2013-01-01

48

Regulation by thyroid hormone of the expression of basement membrane components in rat prepubertal Sertoli cells.  

PubMed

The present study reports the modulation of basement membrane (BM) components, laminin, entactin, and type IV collagen, expression in prepubertal rat Sertoli cell by the thyroid hormone T3. Immunocytochemical studies of permeabilized Sertoli cells in culture showed that T3 treatment (10[-7] M for 24 h) increased the number of cells staining positive for laminin and/or entactin (from 58 +/- 5.3% to 86.4 +/- 6.5%, P < 0.01). In contrast, a strong inhibition of type IV collagen immunopositivity was observed. Western blot analysis of Sertoli cell-conditioned media indicated that T3 treatment significantly (P < 0.01) increased the level of secreted entactin by 60-65% without affecting the levels of laminin A and B1/B2 chains. Moreover, thyroid hormone treatment of Sertoli cells significantly reduced type IV collagen secretion by 62% (P < 0.05). Slot blot analysis of poly-A RNA demonstrated a significant (P < 0.01) increase in the level of entactin messenger RNA (mRNA) by 140% (P < 0.01) and a 50% reduction of type IV collagen alpha1 chain mRNA after thyroid hormone treatment. No effect of the hormone was observed on the accumulation of the laminin B1 and B2 chain mRNAs in Sertoli cell cultures. These effects cannot be ascribed to changes in the degradation of BM components, because no effect of thyroid hormone was observed on plasminogen activators or metalloproteinase secretion by Sertoli cells. These observations indicate the Sertoli cell as a source of entactin within the testis, demonstrate the ability of T3 to differentially regulate the expression of BM components, and can be regarded as a part of the integrated mechanism by which thyroid hormone affects testicular development and differentiation. PMID:9449648

Ulisse, S; Rucci, N; Piersanti, D; Carosa, E; Graziano, F M; Pavan, A; Ceddia, P; Arizzi, M; Muzi, P; Cironi, L; Gnessi, L; D'Armiento, M; Jannini, E A

1998-02-01

49

Altered lipid homeostasis in Sertoli cells stressed by mild hyperthermia.  

PubMed

Spermatogenesis is known to be vulnerable to temperature. Exposures of rat testis to moderate hyperthermia result in loss of germ cells with survival of Sertoli cells (SC). Because SC provide structural and metabolic support to germ cells, our aim was to test the hypothesis that these exposures affect SC functions, thus contributing to germ cell damage. In vivo, regularly repeated exposures (one of 15 min per day, once a day during 5 days) of rat testes to 43 °C led to accumulation of neutral lipids. This SC-specific lipid function took 1-2 weeks after the last of these exposures to be maximal. In cultured SC, similar daily exposures for 15 min to 43 °C resulted in significant increase in triacylglycerol levels and accumulation of lipid droplets. After incubations with [3H]arachidonate, the labeling of cardiolipin decreased more than that of other lipid classes. Another specifically mitochondrial lipid metabolic function, fatty acid oxidation, also declined. These lipid changes suggested that temperature affects SC mitochondrial physiology, which was confirmed by significantly increased degrees of membrane depolarization and ROS production. This concurred with reduced expression of two SC-specific proteins, transferrin, and Wilms' Tumor 1 protein, markers of SC secretion and differentiation functions, respectively, and with an intense SC cytoskeletal perturbation, evident by loss of microtubule network (?-tubulin) and microfilament (f-actin) organization. Albeit temporary and potentially reversible, hyperthermia-induced SC structural and metabolic alterations may be long-lasting and/or extensive enough to respond for the decreased survival of the germ cells they normally foster. PMID:24690895

Vallés, Ana S; Aveldaño, Marta I; Furland, Natalia E

2014-01-01

50

Altered Lipid Homeostasis in Sertoli Cells Stressed by Mild Hyperthermia  

PubMed Central

Spermatogenesis is known to be vulnerable to temperature. Exposures of rat testis to moderate hyperthermia result in loss of germ cells with survival of Sertoli cells (SC). Because SC provide structural and metabolic support to germ cells, our aim was to test the hypothesis that these exposures affect SC functions, thus contributing to germ cell damage. In vivo, regularly repeated exposures (one of 15 min per day, once a day during 5 days) of rat testes to 43°C led to accumulation of neutral lipids. This SC-specific lipid function took 1–2 weeks after the last of these exposures to be maximal. In cultured SC, similar daily exposures for 15 min to 43°C resulted in significant increase in triacylglycerol levels and accumulation of lipid droplets. After incubations with [3H]arachidonate, the labeling of cardiolipin decreased more than that of other lipid classes. Another specifically mitochondrial lipid metabolic function, fatty acid oxidation, also declined. These lipid changes suggested that temperature affects SC mitochondrial physiology, which was confirmed by significantly increased degrees of membrane depolarization and ROS production. This concurred with reduced expression of two SC-specific proteins, transferrin, and Wilms' Tumor 1 protein, markers of SC secretion and differentiation functions, respectively, and with an intense SC cytoskeletal perturbation, evident by loss of microtubule network (?-tubulin) and microfilament (f-actin) organization. Albeit temporary and potentially reversible, hyperthermia-induced SC structural and metabolic alterations may be long-lasting and/or extensive enough to respond for the decreased survival of the germ cells they normally foster.

Valles, Ana S.; Aveldano, Marta I.; Furland, Natalia E.

2014-01-01

51

Androgens and Postmeiotic Germ Cells Regulate Claudin-11 Expression in Rat Sertoli Cells  

Microsoft Academic Search

In the present study we investigated whether fetal exposure to flutamide affected messenger and protein levels of clau- din-11, a key Sertoli cell factor in the establishment of the hemotesticular barrier, at the time of two key events of postnatal testis development: 1) before puberty (postnatal d 14) during the establishment of the hemotesticular barrier, and 2) at the adult

Anne Florin; Magali Maire; Aline Bozec; Ali Hellani; Sonia Chater; Remi Bars; Franck Chuzel; Mohamed Benahmed

2004-01-01

52

Cadmium suppresses the proliferation of piglet Sertoli cells and causes their DNA damage, cell apoptosis and aberrant ultrastructure  

PubMed Central

Objective Very little information is known about the toxic effects of cadmium on somatic cells in mammalian testis. The objective of this study is to explore the toxicity of cadmium on piglet Sertoli cells. Methods Sertoli cells were isolated from piglet testes using a two-step enzyme digestion and followed by differential plating. Piglet Sertoli cells were identified by oil red O staining and Fas ligand (FasL) expression as assayed by immunocytochemistry and expression of transferrin and androgen binding protein by RT-PCR. Sertoli cells were cultured in DMEM/F12 supplemented with 10% fetal calf serum in the absence or presence of various concentrations of cadmium chloride, or treatment with p38 MAPK inhibitor SB202190 and with cadmium chloride exposure. Apoptotic cells in seminiferous tubules of piglets were also performed using TUNEL assay in vivo. Results Cadmium chloride inhibited the proliferation of Piglet Sertoli cells as shown by MTT assay, and it increased malondialdehyde (MDA) but reduced superoxide dismutase (SOD) and Glutathione peroxidase (GSH-Px) activity. Inhibitor SB202190 alleviated the proliferation inhibition of cadmium on piglet Sertoli cells. Comet assay revealed that cadmium chloride caused DNA damage of Piglet Sertoli cells and resulted in cell apoptosis as assayed by flow cytometry. The in vivo study confirmed that cadmium induced cell apoptosis in seminiferous tubules of piglets. Transmission electronic microscopy showed abnormal and apoptotic ultrastructure in Piglet Sertoli cells treated with cadmium chloride compared to the control. Conclusion cadmium has obvious adverse effects on the proliferation of piglet Sertoli cells and causes their DNA damage, cell apoptosis, and aberrant morphology. This study thus offers novel insights into the toxicology of cadmium on male reproduction.

2010-01-01

53

Arylsulfatase A deficiency causes seminolipid accumulation and a lysosomal storage disorder in Sertoli cells[S  

PubMed Central

Sulfogalactosylglycerolipid (SGG) is the major sulfoglycolipid of male germ cells. During spermatogenesis, apoptosis occurs in >50% of total germ cells. Sertoli cells phagocytose these apoptotic germ cells and degrade their components using lysosomal enzymes. Here we demonstrated that SGG was a physiological substrate of Sertoli lysosomal arylsulfatase A (ARSA). SGG accumulated in Sertoli cells of Arsa?/? mice, and at 8 months of age, this buildup led to lysosomal swelling and other cellular abnormalities typical of a lysosomal storage disorder. This disorder likely compromised Sertoli cell functions, manifesting as impaired spermatogenesis and production of sperm with near-zero fertilizing ability in vitro. Fecundity of Arsa?/? males was thus reduced when they were older than 5 months. Sperm SGG is known for its roles in fertilization. Therefore, the minimal sperm fertilizing ability of 8-month-old Arsa?/? males may be explained by the 50% reduction of their sperm SGG levels, a result that was also observed in testicular germ cells. These unexpected decreases in SGG levels might be partly due to depletion of the backbone lipid palmitylpalmitoylglycerol that is generated from the SGG degradation pathway in Sertoli cells and normally recycled to new generations of primary spermatocytes for SGG synthesis.

Xu, Hongbin; Kongmanas, Kessiri; Kadunganattil, Suraj; Smith, Charles E.; Rupar, Tony; Goto-Inoue, Naoko; Hermo, Louis; Faull, Kym F.; Tanphaichitr, Nongnuj

2011-01-01

54

Testes of Astyanax altiparanae: The Sertoli cell functions in a semicystic spermatogenesis.  

PubMed

The Astyanax altiparanae (lambari) is a South American freshwater fish belonging to the family Characidae. Although some authors have described reproductive aspects of this species, this is the first study about the morphology of the testes throughout the annual reproductive cycle of A. altiparanae. Fish spermatogenesis differs from that in mammals as it occurs in cysts whose borders are defined by cytoplasmic processes of Sertoli cells, thus creating a favorable environment for spermatogenesis. The functions commonly attributed to fish Sertoli cells were investigated using stereological, light and electron microscopy in A. altiparanae. Results showed that when the Sertoli cells of A. altiparanae are in contact with germ cells, they plan a support function that culminates in the production of spermatozoa. After releasing spermatozoa, modified Sertoli cells form the duct epithelium, transform into secretory cells and release a secretion into the duct lumen where spermatids and sperm are located. Thus, the present study revealed important aspects of the testes of A. altiparanae, and propose a sequence of functions played by the Sertoli cells in this species. PMID:24792443

Costa, F G; Adolfi, M C; Gomes, C C; Jesus, L W O; Batlouni, S R; Borella, M I

2014-06-01

55

Germ cell control of testin production is inverse to that of other Sertoli cell products.  

PubMed

Recent studies have shown that germ cells can regulate testins, two newly identified Sertoli cell proteins that are associated with junctional complexes. To investigate this possibility, several parameters of Sertoli cell function were investigated over 2-120 days post exposure of the rat testes to x-rays (3 Grays). The irradiation-induced loss of spermatogonia resulted in a maturation-depletion process progressively affecting all germ cell classes. Testis weight began to decrease when the most numerous germ cell type (spermatids) began to decline. A complete or near complete recovery of spermatogenesis and of the testis weight had occurred by day 120 post irradiation. There was no significant change in FSH, epididymal androgen-binding protein, and tubule fluid levels during the first weeks after irradiation, when the seminiferious epithelium was depleted of spermatogonia and germ cells up to early spermatids. In contrast, when the number of the more mature forms of spermatids declined (between day 21 and 54), FSH rose and androgen-binding protein as well as fluid production declined. The subsequent recovery of these parameters was also highly correlated with the number of late spermatids. By contrast, testicular testin contents reacted to the depletion of germ cells with a biphasic increase; a doubling occurred when spermatogonia, spermatocytes, and early spermatids were absent (days 4-28), and a 7-fold rise occurred by day 37 when the number of late spermatids had decreased by 50%. By day 54, when the sperm counts had reached a nadir, testin contents had returned to levels corresponding to about four times the control levels; they progressively recovered thereafter. These observations support the postulate that germ cells negatively regulate testins. This possibility was investigated with in vitro experiments showing that addition of germ cell-conditioned medium to Sertoli cell monolayers inhibited testin secretion in a dose-dependent manner. In conclusion this study; 1) highlights the complex interplay between the various germ cell classes in the control of the Sertoli cell function in the adult testis; 2) establishes that germ cell effects may be opposite on different Sertoli cell products; 3) demonstrates that several classes of germ cells negatively control testicular testin contents; and 4) emphasizes the particular role of late spermatids in Sertoli cell regulation. PMID:8504757

Jégou, B; Pineau, C; Velez de la Calle, J F; Touzalin, A M; Bardin, C W; Cheng, C Y

1993-06-01

56

The Sertoli cell junctional specializations and their relationship to the germinal epithelium as observed after efferent ductule ligation.  

PubMed

Seminiferous tubules from testes of normal and efferent ductule ligated mice were examined with the electron microscope. The tubules in the ligated animals were markedly distended and at most stages of the seminiferous cycle the epithelium exhibited a series of circumferentially-oriented ridges. Cross-sectional profiles of these ridges were studied with particular emphasis on the Sertoli cell junctional specializations and their relationship to the germinal cells. In the ligated specimen the basal cytoplasm of the Sertoli cells is highly attenuated, often appearing as a thin process resting on the basement lamina. Where the cytoplasm of one Sertoli cell ends, it meets in apposition with the cytoplasm of an adjoining Sertoli cell, and at these sites, junctional specializations are present. The ridges are comprised of a stalk of apical Sertoli cell cytoplasm, often appearing like an inverted cone, with young spermatids aligned along the lateral surfaces and the more mature spermatid population embedded within the apical cytoplasm. Junctional specializations were observed along these lateral Sertoli cell surfaces. In some instances, they formed a free surface, but usually early spermatids were in contact with the junctional specializations. With respect to the more mature spermatids, the acrosomal component was typically found in relation to a junctional specialization. Germ cells at the spermatocyte stage were also noted in relation to the Sertoli cell junctional specializations. The findings suggest that spermatocytes cross the Sertoli cell barrier and gain access to the adluminal compartment of the seminiferous tubule through the disengagement of the inter-Sertoli cell junctional complex. It is proposed that when the inter-Sertoli cell junctional specializations separate, the spermatocytes come in apposition with the newly freed junctional surfaces and remain in relation with them through the ensuing divisions. It appears that at some point, firm adhesion between germ cells and the junctional specializations occurs; the spermatid progeny may thus maintain contact with the original inter-Sertoli cell junctional specializations until their release into the tubule lumen. PMID:1200403

Ross, M H; Dobler, J

1975-10-01

57

[Effects of anti-cancer agents on cultured rat Sertoli cells].  

PubMed

We performed primary immature Sertoli cell culture to investigate whether or not anti-cancer agents would have a direct effect on rat Sertoli cells. Sertoli cells, isolated from testes of 18-day-old rats, were cultured in pellets with medium for 5 days. The concentration of transferrin in cultured medium were measured as the function of Sertoli cells. The anti-cancer agents cis-diamminedichloroplatinum (CDDP), adriamycin and vinblastine were selected for this study, and added to the culture medium. CDDP decreased the level of transferrin concentration in cultured medium, namely 0.5 microgram/ml of CDDP resulted in 54.9% of the transferrin concentration in the medium compared with that without any anti-cancer agents (p < 0.05), and 1.0 micrograms/ml of CDDP produced transferrin concentrations of 57.5% and 46.2% (p < 0.05), respectively. Adriamycin (0.4 microgram/ml) and vinblastine (0.5 microgram/ml) produced transferrin concentrations of 35.2% and 31.3% in cultured medium (p < 0.05), respectively. These findings revealed that anti-cancer agents have direct damaging effects on rat Sertoli cells. PMID:7609355

Nambu, A; Kumamoto, Y; Mikuma, N

1995-06-01

58

Effects of PFNA exposure on expression of junction-associated molecules and secretory function in rat Sertoli cells  

Microsoft Academic Search

Perfluorononanoic acid (PFNA, C9), a synthetic perfluorinated chemical containing nine carbons, has been identified in humans and wildlife worldwide. Sertoli cell plays a key role in spermatogenesis; however, the toxic effects of PFNA on Sertoli cells have not been studied. The aim of this study was to investigate the effects of PFNA exposure on cell junction molecules and factors specifically

Yixing Feng; Xuemei Fang; Zhimin Shi; Muqi Xu; Jiayin Dai

2010-01-01

59

Selective Ablation of the Androgen Receptor in Mouse Sertoli Cells Affects Sertoli Cell Maturation, Barrier Formation and Cytoskeletal Development  

PubMed Central

The observation that mice with a selective ablation of the androgen receptor (AR) in Sertoli cells (SC) (SCARKO mice) display a complete block in meiosis supports the contention that SC play a pivotal role in the control of germ cell development by androgens. To delineate the physiological and molecular mechanism responsible for this control, we compared tubular development in pubertal SCARKO mice and littermate controls. Particular attention was paid to differences in SC maturation, SC barrier formation and cytoskeletal organization and to the molecular mediators potentially involved. Functional analysis of SC barrier development by hypertonic perfusion and lanthanum permeation techniques and immunohistochemical analysis of junction formation showed that SCARKO mice still attempt to produce a barrier separating basal and adluminal compartment but that barrier formation is delayed and defective. Defective barrier formation was accompanied by disturbances in SC nuclear maturation (immature shape, absence of prominent, tripartite nucleoli) and SC polarization (aberrant positioning of SC nuclei and cytoskeletal elements such as vimentin). Quantitative RT-PCR was used to study the transcript levels of genes potentially related to the described phenomena between day 8 and 35. Differences in the expression of SC genes known to play a role in junction formation could be shown from day 8 for Cldn11, from day 15 for Cldn3 and Espn, from day 20 for Cdh2 and Jam3 and from day 35 for ZO-1. Marked differences were also noted in the transcript levels of several genes that are also related to cell adhesion and cytoskeletal dynamics but that have not yet been studied in SC (Actn3, Ank3, Anxa9, Scin, Emb, Mpzl2). It is concluded that absence of a functional AR in SC impedes the remodeling of testicular tubules expected at the onset of spermatogenesis and interferes with the creation of the specific environment needed for germ cell development.

Willems, Ariane; Batlouni, Sergio R.; Esnal, Arantza; Swinnen, Johannes V.; Saunders, Philippa T. K.; Sharpe, Richard M.; Franca, Luiz R.; De Gendt, Karel; Verhoeven, Guido

2010-01-01

60

Inhibin-? immunohistochemical expression in mature and immature canine Sertoli and Leydig cells.  

PubMed

Formalin-fixed, paraffin-embedded sections from 32 canine pairs of testes were immunohistochemically examined for Inhibin-? (INH?). Samples were subdivided into two groups (group 1, neonates; group 2, puppies and adults) and results statistically compared. Inhibin-? was significantly expressed only in Sertoli cells of neonatal testes, while in Leydig cells it was expressed without significant difference between groups. These results suggest that, in dogs, INH? expression switches from Sertoli to Leydig cells during testicular maturation and that, in adult, Leydig cells represent the main source of INH?. PMID:21492261

Grieco, V; Banco, B; Ferrari, A; Rota, A; Faustini, M; Veronesi, M C

2011-10-01

61

Retinoic acid metabolism links the periodical differentiation of germ cells with the cycle of Sertoli cells in mouse seminiferous epithelium.  

PubMed

Homeostasis of tissues relies on the regulated differentiation of stem cells. In the epithelium of mouse seminiferous tubules, the differentiation process from undifferentiated spermatogonia (A(undiff)), which harbor the stem cell functions, to sperm occurs in a periodical manner, known as the "seminiferous epithelial cycle". To identify the mechanism underlying this periodic differentiation, we investigated the roles of Sertoli cells (the somatic supporting cells) and retinoic acid (RA) in the seminiferous epithelial cycle. Sertoli cells cyclically change their functions in a coordinated manner with germ cell differentiation and support the entire process of spermatogenesis. RA is known to play essential roles in this periodic differentiation, but its precise mode of action and its regulation remains largely obscure. We showed that an experimental increase in RA signaling was capable of both inducing A(undiff) differentiation and resetting the Sertoli cell cycle to the appropriate stage. However, these actions of exogenous RA signaling on A(undiff) and Sertoli cells were strongly interfered by the differentiating germ cells of intimate location. Based on the expression of RA metabolism-related genes among multiple cell types - including germ and Sertoli cells - and their regulation by RA signaling, we propose here that differentiating germ cells play a primary role in modulating the local RA metabolism, which results in the timed differentiation of A(undiff) and the appropriate cycling of Sertoli cells. Similar regulation by differentiating progeny through the modulation of local environment could also be involved in other stem cell systems. PMID:22200512

Sugimoto, Ryo; Nabeshima, Yo-ichi; Yoshida, Shosei

2012-01-01

62

Somatostatin and its receptors: functional regulation in the development of mice Sertoli cells.  

PubMed

Recently, Sertoli cells have been ascertained as the target for the regulatory peptide somatostatin (SST). Therefore, the present study investigated the expression of somatostatin receptors, their age-related alterations and homologous regulation by in vitro treatment with SRIF14 on mice Sertoli cells; furthermore, it dealt with SRIF14 action on growth progression, apoptotic activity and related gene expressions in these cells. We found that mice Sertoli cells expressed all SST1-5 receptors with differential intensities. Age-related real-time PCR of all somatostatin receptors identified abundance of SSTR2 and SSTR5 mRNA level during Sertoli cell developmental period. Furthermore, higher level of these two receptors was independent of SRIF14, as treatment with SRIF14 failed to induce both receptor expressions when compared with control. Somatostatin treatment elicited a dose-dependent decrease in forskolin stimulated cAMP production. Low (100pM and 10nM) and high dosage (1?M) groups of SRIF14 significantly promoted apoptosis, while all treatment groups led to dose dependent cessation (P<0.05) of G1 phase of cell cycle as further validated by increase in casp3, decrease in bcl2, elevation of P21 (all by western blot) and decrease in Igf1 expressions, similarly, SST treatment caused a dose dependent suppression in the mRNA level of kitl gene, which is important in the regulation of spermatogenesis. These findings suggest that somatostatin and its receptors (SSTR2 and SSTR5) are important markers in the regulation and development of Sertoli cell; furthermore, it portrays physiological inhibitory role in Sertoli cell development by inducing apoptosis and cell cycle arrest. PMID:23831358

Riaz, Hasan; Liang, Aixin; Khan, Muhammad Kasib; Dong, Ping; Han, Li; Shahzad, Muhammad; Chong, Zhenlu; Ahmad, Sibtain; Hua, Guohua; Yang, Liguo

2013-11-01

63

Icariin stimulates the proliferation of rat Sertoli cells in an ERK1/2-dependent manner in vitro.  

PubMed

Icariin (ICA), a major constituent of flavonoids from the Chinese medical herb Epimedium brevicornum Maxim, is found to be protective for male reproductive ability, with the underlying mechanism largely unknown. Our study here investigated the effects of ICA on Sertoli cells, which act as nurse cells for the germ cells developing. Icariin was found to stimulate Sertoli cell proliferation in a dose-dependent manner. Further study revealed that Icariin induced an obvious phosphorylation of ERK in Sertoli cells. Inhibition of activation of ERK by the ERK inhibitor U0126 nearly blocked the Icariin-induced proliferation of Sertoli cells. Taken together, our results suggest that Icariin promotes the proliferation of Sertoli cells in vitro by activating the ERK1/2 signal pathway, which might at least partially, explain the protective role of Icariin on male reproductive ability. PMID:23134192

Nan, Y; Zhang, X; Yang, G; Xie, J; Lu, Z; Wang, W; Ni, X; Cao, X; Ma, J; Wang, Z

2012-11-01

64

Initiation of testicular tubulogenesis is controlled by neurotrophic tyrosine receptor kinases in a three-dimensional Sertoli cell aggregation assay.  

PubMed

The first morphological sign of testicular differentiation is the formation of testis cords. Prior to cord formation, newly specified Sertoli cells establish adhesive junctions, and condensation of somatic cells along the surface epithelium of the genital ridge occurs. Here, we show that Sertoli cell aggregation is necessary for subsequent testis cord formation, and that neurotrophic tyrosine kinase receptors (NTRKs) regulate this process. In a three-dimensional cell culture assay, immature rat Sertoli cells aggregate to form large spherical aggregates (81.36+/-7.34 microm in diameter) in a highly organized, hexagonal arrangement (376.95+/-21.93 microm average distance between spherical aggregates). Exposure to NTRK inhibitors K252a and AG879 significantly disrupted Sertoli cell aggregation in a dose-dependent manner. Sertoli cells were prevented from establishing cell-cell contacts and from forming spherical aggregates. In vitro-derived spherical aggregates were xenografted into immunodeficient nude mice to investigate their developmental potential. In controls, seminiferous tubule-like structures showing polarized single-layered Sertoli cell epithelia, basement membranes, peritubular myoid cells surrounding the tubules, and lumen were observed in histological sections. By contrast, grafts from treatment groups were devoid of tubules and only few single Sertoli cells were present in xenografts after 4 weeks. Furthermore, the grafts were significantly smaller when Sertoli cell aggregation was disrupted by K252a in vitro (20.87 vs 6.63 mg; P<0.05). We conclude from these results that NTRK-regulated Sertoli-Sertoli cell contact is essential to the period of extensive growth and remodeling that occurs during testicular tubulogenesis, and our data indicate its potential function in fetal and prepubertal testis differentiation. PMID:18660385

Gassei, Kathrin; Ehmcke, Jens; Schlatt, Stefan

2008-10-01

65

The kinase DYRKIA regulates pre-mRNA splicing in spermatogonia and proliferation of spermatogonia and Sertoli cells by phosphorylating a spliceosomal component, SAP155, in postnatal murine testes  

Microsoft Academic Search

SAP155 is an essential component of the spliceosome and its phosphorylation is required for splicing catalysis, but little\\u000a is known concerning its function and regulation during spermatogenesis in postnatal murine testes. We report that inhibition\\u000a of dual-specificity tyrosine-phosphorylation regulated kinase (DYRK) IA strongly suppressed the mitogen-stimulated SAP155\\u000a phosphorylation and constitutive splicing of I?B pre-mRNA as well as the proliferation of

Ko EtoYoshiyuki; Yoshiyuki Sonoda; Shin-ichi Abe

2011-01-01

66

Mono(2-ethylhexyl) Phthalate Rapidly Alters both Sertoli Cell Vimentin Filaments and Germ Cell Apoptosis in Young Rat Testes  

Microsoft Academic Search

Mono-(2-ethylhexyl) phthalate (MEHP) is a widely studied Sertoli cell toxicant. Here we describe alterations in Sertoli cell vimentin filament distribution and the incidence of testicular germ cell apoptosis in young (28-day-old) Fischer rats that were treated with MEHP (2 g\\/kg, po) and killed 0, 3, 6, or 12 hr after exposure. A collapse in vimentin filaments was observed 3 hr

John H. Richburg; Kim Boekelheide

1996-01-01

67

Growth differentiation factor 9 is a germ cell regulator of Sertoli cell function.  

PubMed

Oocyte-secreted growth differentiation factor (GDF) 9 and bone morphogenetic protein (BMP) 15 are critical regulatory factors in female reproduction. Together, they promote granulosa cell proliferation and stimulate the maturation of preovulatory follicles. Despite their importance in female fertility, GDF9 and BMP15 expression patterns and function during spermatogenesis have not been investigated. In this study we show that the expression and stage-specific localization of both factors are limited to the germ cells of the rat seminiferous epithelium, with GDF9 being principally localized in round spermatids and BMP15 in gonocytes and pachytene spermatocytes. To identify potential cellular targets for GDF9 actions, cells of the seminiferous tubule were isolated and screened for the expression of signaling receptors [activin-like kinase (ALK) 5, ALK6, and BMP receptor, type II)]. Individual receptor types were expressed throughout the seminiferous epithelium, but coexpression of ALK5 and BMP receptor, type II was limited to Sertoli cells and round spermatids. Based on the reproductive actions of related TGFbeta ligands in the ovary and testis, GDF9 was assessed for its ability to regulate tight junction function and inhibin B production in rat Sertoli cell cultures. When recombinant mouse GDF9 was added to immature Sertoli cell cultures, it inhibited membrane localization of the junctional proteins claudin-11, occludin, and zonula occludens-1, thereby disrupting tight junction integrity. Concomitantly, GDF9 up-regulated inhibin subunit expression and significantly stimulated dimeric inhibin B protein production. Together, these results demonstrate that GDF9 and BMP15 are germ cell-specific factors in the rat testis, and that GDF9 can modulate key Sertoli cell functions. PMID:19106224

Nicholls, Peter K; Harrison, Craig A; Gilchrist, Robert B; Farnworth, Paul G; Stanton, Peter G

2009-05-01

68

Lipopolysaccharide inhibits the self-renewal of spermatogonial stem cells in vitro via downregulation of GDNF expression in Sertoli cells.  

PubMed

Lipopolysaccharide (LPS) can reduce sperm count and sperm quality. The molecular mechanisms underlying this process are not fully understood. In this report, we investigated the effects of LPS-treated Sertoli cells on self-renewal and differentiation of spermatogoinial stem cells (SSCs). Sertoli cell cultures were established and incubated with LPS (10?g/ml) for 1, 2 or 3 days, respectively. The culture media were collected and used as conditioned media (CM) to culture SSCs. The expression of glial cell-derived neurotrophic factor (GDNF), stem cell factor (SCF) and bone morphogenetic protein 4 (BMP4) in Sertoli cells treated with LPS was analyzed by RT-PCR and Western blotting. The results showed that the expression of SSC differentiation markers, c-kit and Sohlh2, was increased, while the expression of SSC self-renewal markers, plzf, oct4, and PCNA, was repressed when cultured in CM from LPS-treated Sertoli cells. GDNF levels in Sertoli cells and CM reduced dramatically after LPS treatments, while SCF and BMP4 levels did not show any significant changes. Moreover, correlated with the GDNF levels in CM, GDNF target genes, Bcl6b and Etv5, were reduced markedly in SSCs. Our results suggest that LPS inhibits the expression of GDNF in Sertoli cells, and might prevent the SSC self-renewal via down-regulation of GDNF target genes. PMID:24503145

Zhang, Xiaoli; Shi, Kun; Li, Yi; Zhang, Haiyu; Hao, Jing

2014-06-01

69

Nesprin-3 connects plectin and vimentin to the nuclear envelope of Sertoli cells but is not required for Sertoli cell function in spermatogenesis.  

PubMed

Nesprin-3 is a nuclear envelope protein that connects the nucleus to intermediate filaments by interacting with plectin. To investigate the role of nesprin-3 in the perinuclear localization of plectin, we generated nesprin-3-knockout mice and examined the effects of nesprin-3 deficiency in different cell types and tissues. Nesprin-3 and plectin are coexpressed in a variety of tissues, including peripheral nerve and muscle. The expression level of nesprin-3 in skeletal muscle is very low and decreases during myoblast differentiation in vitro. Of interest, plectin was concentrated at the nuclear envelope in only a few cell types. This was most prominent in Sertoli cells of the testis, in which nesprin-3 is required for the localization of both plectin and vimentin at the nuclear perimeter. Testicular morphology and the position of the nucleus in Sertoli cells were normal, however, in the nesprin-3-knockout mice and the mice were fertile. Furthermore, nesprin-3 was not required for the polarization and migration of mouse embryonic fibroblasts. Thus, although nesprin-3 is critical for the localization of plectin to the nuclear perimeter of Sertoli cells, the resulting link between the nuclear envelope and the intermediate filament system seems to be dispensable for normal testicular morphology and spermatogenesis. PMID:23761073

Ketema, Mirjam; Kreft, Maaike; Secades, Pablo; Janssen, Hans; Sonnenberg, Arnoud

2013-08-01

70

Structural response of the hamster Sertoli cell to hypophysectomy: a correlative morphometric and endocrine study.  

PubMed

Reproductively active hamsters were hypophysectomized and examined 6 or 20 days later in a combined morphometric and endocrine study of the Sertoli cell to determine 1) the morphological and endocrine effects of hypophysectomy of both short- and long-term duration, 2) if regression of Sertoli cells after hypophysectomy in a seasonal breeder resembles regression due to seasonal changes, and 3) if effects of hypophysectomy in a seasonal breeder are equivalent to the effects of hypophysectomy in a nonseasonal breeder. Six days after hypophysectomy, at a period when germ cell degeneration is first noted, there was a significant decrease in testis weight, interstitial space, tubule diameter and length, volume of seminiferous tubule, and tubular lumen. There were no significant changes in Sertoli cell nuclear and cytoplasmic volume although cell surface area was decreased significantly. Most organelles exhibited no significant change in volume or surface area except for secondary lysosomes which expectedly increased in volume as the result of phagocytosis of germinal cells. Thus at an early time period when functional changes in germ cells and Leydig cells are clearly evident (Russell et al. [1992] Endocrinology), the Sertoli cell shows minimal changes. Twenty days after hypophysectomy, the cell, nuclear and cytoplasmic volumes and surface area of the Sertoli cells, and volumes and surface areas of nearly all organelles were significantly decreased from values measured in normal and in short-term hypophysectomized hamsters. The exceptions were the total volumes of lipid which increased significantly and lysosomes which were similar to normal but significantly lower than short-term hypophysectomized animals. The long-term hypophysectomized hamster Sertoli cell, like that of the short-day hamster (Sinha Hikim et al. [1989b] Endocrinology, 125:1829-1843) is structurally regressed as a whole rather than exhibiting selective decreases in cellular and subcellular components. The size of the Sertoli cell in pituitary-intact, long- and short-term hypophysectomized animals showed positive and significant correlations with the volumes and surface areas of all its cytoplasmic organelles except the volume of lipid which showed a negative, significant correlation. Comparisons of long-term hypophysectomized hamsters (in long-day light exposure) and short-day exposed animals (Sinha Hikim et al. [1989b] (Endocrinology, 125:1829-1843) suggested that hypophysectomy, in general, resulted in similar, but slightly more severe regressive changes in the testis and germ cell population than those seen during seasonal regression.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:1456454

Ghosh, S; Bartke, A; Grasso, P; Reichert, L E; Russell, L D

1992-12-01

71

Protein secretory patterns of rat Sertoli and peritubular cells are influenced by culture conditions  

SciTech Connect

An approach combining two-dimensional gel electrophoresis and autoradiography was used to correlate patterns of secretory proteins in cultures of Sertoli and peritubular cells with those observed in the incubation medium from segments of seminiferous tubules. Sertoli cells in culture and in seminiferous tubules secreted three proteins designated S70 (Mr 72,000-70,000), S45 (Mr 45,000), and S35 (Mr 35,000). Cultured Sertoli and peritubular cells and incubated seminiferous tubules secreted two proteins designated SP1 (Mr 42,000) and SP2 (Mr 50,000). SP1 and S45 have similar Mr but differ from each other in isoelectric point (pI). Cultured peritubular cells secreted a protein designated P40 (Mr 40,000) that was also seen in intact seminiferous tubules but not in seminiferous tubules lacking the peritubular cell wall. However, a large number of high-Mr proteins were observed only in the medium of cultured peritubular cells but not in the incubation medium of intact seminiferous tubules. Culture conditions influence the morphology and patterns of protein secretion of cultured peritubular cells. Peritubular cells that display a flat-stellate shape transition when placed in culture medium free of serum (with or without hormones and growth factors), accumulate various proteins in the medium that are less apparent when these cells are maintained in medium supplemented with serum. Two secretory proteins stimulated by follicle-stimulating hormone (FSH) (designated SCm1 and SCm2) previously found in the medium of cultured Sertoli cells, were also observed in the incubation medium of seminiferous tubular segments stimulated by FSH. Results of this study show that, although cultured Sertoli and peritubular cells synthesize and secrete proteins also observed in segments of incubated seminiferous tubules anther group of proteins lacks seminiferous tubular correlates.

Kierszenbaum, A.L.; Crowell, J.A.; Shabanowitz, R.B.; DePhilip, R.M.; Tres, L.L.

1986-08-01

72

Analysis of Sertoli cell efficiency allows the differentiation between two fundamentally different forms of feline teratospermia.  

PubMed

Teratospermia is a common phenomenon within felid species and has been attributed to reduction in genetic diversity. Testes from teratospermic domestic cats show enhanced spermatogenesis accompanied by remarkably reduced germ cell apoptosis. In the present study we investigated whether free-range teratospermic tom cats exhibit a similar testicular phenotype as proven permanently teratospermic males. Randomly collected teratospermic cats were compared with normal (normospermic; >60% morphologically normal sperm per ejaculate) and a well-characterized population of permanently teratospermic domestic cats, with respect to their spermatogenic potential. Histomorphologic assessment of testes from randomly collected teratospermic cats revealed no differences compared with normospermic donors. These two groups, however, were both different from permanently teratospermic cats, which exhibit fewer Sertoli cells and increased numbers of round spermatids per tubule cross-section resulting in a remarkably increased Sertoli cell efficiency (ratio of round spermatids to Sertoli cells). In conclusion, we can distinguish at least two fundamentally different forms of feline teratospermia. One subtype, found in most of the randomly collected tom cats, but not associated with altered quantitative spermatogenic parameters. Another subtype, found in all permanently teratospermic felids, is manifested by an impairment of Sertoli cell efficiency. We suggest that spermatogenic output should be analyzed before using random source domestic cats to study the phenomenon of teratospermia. PMID:23174773

Jewgenow, K; Pukazhenthi, B S; Schoen, J

2013-01-15

73

Androgen-dependent sertoli cell tight junction remodeling is mediated by multiple tight junction components.  

PubMed

Sertoli cell tight junctions (SCTJs) of the seminiferous epithelium create a specialized microenvironment in the testis to aid differentiation of spermatocytes and spermatids from spermatogonial stem cells. SCTJs must be chronically broken and rebuilt with high fidelity to allow the transmigration of preleptotene spermatocytes from the basal to adluminal epithelial compartment. Impairment of androgen signaling in Sertoli cells perturbs SCTJ remodeling. Claudin (CLDN) 3, a tight junction component under androgen regulation, localizes to newly forming SCTJs and is absent in Sertoli cell androgen receptor knockout (SCARKO) mice. We show here that Cldn3-null mice do not phenocopy SCARKO mice: Cldn3(-/-) mice are fertile, show uninterrupted spermatogenesis, and exhibit fully functional SCTJs based on imaging and small molecule tracer analyses, suggesting that other androgen-regulated genes must contribute to the SCARKO phenotype. To further investigate the SCTJ phenotype observed in SCARKO mutants, we generated a new SCARKO model and extensively analyzed the expression of other tight junction components. In addition to Cldn3, we identified altered expression of several other SCTJ molecules, including down-regulation of Cldn13 and a noncanonical tight junction protein 2 isoform (Tjp2iso3). Chromatin immunoprecipitation was used to demonstrate direct androgen receptor binding to regions of these target genes. Furthermore, we demonstrated that CLDN13 is a constituent of SCTJs and that TJP2iso3 colocalizes with tricellulin, a constituent of tricellular junctions, underscoring the importance of androgen signaling in the regulation of both bicellular and tricellular Sertoli cell tight junctions. PMID:24825397

Chakraborty, Papia; William Buaas, F; Sharma, Manju; Smith, Benjamin E; Greenlee, Anne R; Eacker, Stephen M; Braun, Robert E

2014-07-01

74

Loss of Dicer in Sertoli Cells Has a Major Impact on the Testicular Proteome of Mice  

PubMed Central

Sertoli cells (SCs) are the central, essential coordinators of spermatogenesis, without which germ cell development cannot occur. We previously showed that Dicer, an RNaseIII endonuclease required for microRNA (miRNA) biogenesis, is absolutely essential for Sertoli cells to mature, survive, and ultimately sustain germ cell development. Here, using isotope-coded protein labeling, a technique for protein relative quantification by mass spectrometry, we investigated the impact of Sertoli cell-Dicer and subsequent miRNA loss on the testicular proteome. We found that, a large proportion of proteins (50 out of 130) are up-regulated by more that 1.3-fold in testes lacking Sertoli cell-Dicer, yet that this protein up-regulation is mild, never exceeding a 2-fold change, and is not preceeded by alterations of the corresponding mRNAs. Of note, the expression levels of six proteins of interest were further validated using the Absolute Quantification (AQUA) peptide technology. Furthermore, through 3?UTR luciferase assays we identified one up-regulated protein, SOD-1, a Cu/Zn superoxide dismutase whose overexpression has been linked to enhanced cell death through apoptosis, as a likely direct target of three Sertoli cell-expressed miRNAs, miR-125a-3p, miR-872 and miR-24. Altogether, our study, which is one of the few in vivo analyses of miRNA effects on protein output, suggests that, at least in our system, miRNAs play a significant role in translation control.

Papaioannou, Marilena D.; Lagarrigue, Melanie; Vejnar, Charles E.; Rolland, Antoine D.; Kuhne, Francoise; Aubry, Florence; Schaad, Olivier; Fort, Alexandre; Descombes, Patrick; Neerman-Arbez, Marguerite; Guillou, Florian; Zdobnov, Evgeny M.; Pineau, Charles; Nef, Serge

2011-01-01

75

Morphological and histometric study on the human Sertoli cell from birth to the onset of puberty.  

PubMed Central

In order to evaluate the changes in the number and form of the Sertoli cell from birth to the onset of puberty, a histometric and ultrastructural study was carried out in normal children. Ultrastructural findings revealed that immature Sertoli cells, present from birth to puberty, show round to elliptical nuclei, with regular outlines and small nucleoli. The cytoplasm exhibits well developed rough endoplasmic reticulum, prominent supranuclear Golgi complexes and vesicles at the luminal face; smooth endoplasmic reticulum was scarce. Histometric study revealed a progressive decrease in the Sertoli cell number per transverse tubular section as well as per unit area of testicular parenchyma, mainly from 3 years onwards. However, this decrease seemed to be the result of the progressive increase in the testicular volume as well as in the length and width of the seminiferous tubules, without change in the total Sertoli cell number per testis. This number may be considered constant, not only for adult testes but also for postnatal developing testes. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5

Nistal, M; Abaurrea, M A; Paniagua, R

1982-01-01

76

Dynamin 2 is required for actin assembly in phagocytosis in Sertoli cells  

SciTech Connect

Dynamin 2 has been reported to be implicated in phagocytosis. However, the mode of action of dynamin is poorly understood. In this study, we examined whether dynamin 2 participates in actin assembly during phagocytosis in Sertoli cells. In the presence of dynasore, a dynamin inhibitor, phagocytosis was reduced by 60-70% in Sertoli cells and macrophages. Scanning electron microscopy revealed that Sertoli cells treated with dynasore were unable to form phagocytic cups. In addition, dysfunction of dynamin 2 reduced both actin polymerization and recruitment of actin and dynamin 2 to phosphatidylinositol (4,5) bisphosphate [PI(4,5)P{sub 2}]-containing liposomes. The formation of dynamin 2-positive ruffles of Sertoli cells was decreased by 60-70% by sequestering PI(4,5)P{sub 2} either by expression of PH domain of PLC{delta} or treatment with neomycin. These results strongly suggest that dynamin 2 is involved in actin dynamics and the formation of dynamin 2-positive ruffles during phagocytosis.

Otsuka, Atsushi; Abe, Tadashi [Department of Neuroscience, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558 (Japan); Watanabe, Masami [Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558 (Japan); Yagisawa, Hitoshi [Laboratory of Biological Signaling, Graduate School of Life Science, University of Hyogo, Harima Science Garden City, Kouto 3-chome, Kamigori, Hyogo 678-1297 (Japan); Takei, Kohji [Department of Neuroscience, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558 (Japan); Yamada, Hiroshi [Department of Neuroscience, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558 (Japan)], E-mail: hiroyama@md.okayama-u.ac.jp

2009-01-16

77

Basolateral Uptake of Nucleosides by Sertoli Cells Is Mediated Primarily by Equilibrative Nucleoside Transporter 1  

PubMed Central

The blood-testis barrier (BTB) prevents the entry of many xenobiotic compounds into seminiferous tubules thereby protecting developing germ cells. Understanding drug transport across the BTB may improve drug delivery into the testis. Members of one class of drug, nucleoside reverse transcriptase inhibitors (NRTIs), do penetrate the BTB, presumably through interaction with physiologic nucleoside transporters. By investigating the mechanism of nucleoside transport, it may be possible to design other drugs to bypass the BTB in a similar manner. We present a novel ex vivo technique to study transport at the BTB that employs isolated, intact seminiferous tubules. Using this system, we found that over 80% of total uptake by seminiferous tubules of the model nucleoside uridine could be inhibited by 100 nM nitrobenzylmercaptopurine riboside (NBMPR, 6-S-[(4-nitrophenyl)methyl]-6-thioinosine), a concentration that selectively inhibits equilibrative nucleoside transporter 1 (ENT1) activity. In primary cultured rat Sertoli cells, 100 nM NBMPR inhibited all transepithelial transport and basolateral uptake of uridine. Immunohistochemical staining showed ENT1 to be located on the basolateral membrane of human and rat Sertoli cells, whereas ENT2 was located on the apical membrane of Sertoli cells. Transepithelial transport of uridine by rat Sertoli cells was partially inhibited by the NRTIs zidovudine, didanosine, and tenofovir disoproxil fumarate, consistent with an interaction between these drugs and ENT transporters. These data indicate that ENT1 is the primary route for basolateral nucleoside uptake into Sertoli cells and a possible mechanism for nucleosides and nucleoside-based drugs to undergo transepithelial transport.

Klein, David M.; Evans, Kristen K.; Hardwick, Rhiannon N.; Dantzler, William H.; Wright, Stephen H.

2013-01-01

78

CTNNB1 Signaling in Sertoli Cells Downregulates Spermatogonial Stem Cell Activity via WNT4  

PubMed Central

Constitutive activation of the WNT signaling effector CTNNB1 (?-catenin) in the Sertoli cells of the Ctnnb1tm1Mmt/+;Amhr2tm3(cre)Bhr/+ mouse model results in progressive germ cell loss and sterility. In this study, we sought to determine if this phenotype could be due to a loss of spermatogonial stem cell (SSC) activity. Reciprocal SSC transplants between Ctnnb1tm1Mmt/+;Amhr2tm3(cre)Bhr/+ and wild-type mice showed that SSC activity is lost in Ctnnb1tm1Mmt/+;Amhr2tm3(cre)Bhr/+ testes over time, whereas the mutant testes could not support colonization by wild-type SSCs. Microarray analyses performed on cultured Sertoli cells showed that CTNNB1 induces the expression of genes associated with the female sex determination pathway, which was also found to occur in Ctnnb1tm1Mmt/+;Amhr2tm3(cre)Bhr/+ testes. One CTNNB1 target gene encoded the secreted signaling molecule WNT4. We therefore tested the effects of WNT4 on SSC-enriched germ cell cultures, and found that WNT4 induced cell death and reduced SSC activity without affecting cell cycle. Conversely, conditional inactivation of Wnt4 in the Ctnnb1tm1Mmt/+;Amhr2tm3(cre)Bhr/+ model rescued spermatogenesis and male fertility, indicating that WNT4 is the major effector downstream of CTNNB1 responsible for germ cell loss. Furthermore, WNT4 was found to signal via the CTNNB1 pathway in Sertoli cells, suggesting a self-reinforcing positive feedback loop. Collectively, these data indicate for the first time that ectopic activation of a signaling cascade in the stem cell niche depletes SSC activity through a paracrine factor. These findings may provide insight into the pathogenesis of male infertility, as well as embryonic gonadal development.

Boyer, Alexandre; Yeh, Jonathan R.; Zhang, Xiangfan; Paquet, Marilene; Gaudin, Aurore; Nagano, Makoto C.; Boerboom, Derek

2012-01-01

79

Endosulfan induced cell death in Sertoli-germ cells of male Wistar rat follows intrinsic mode of cell death.  

PubMed

Health of germ cells may affect production of quality gametes either due to endogenous or exogenous factors. Pesticides are among the exogenous factors that can enter the organisms through various routes of exposure and also can affect the reproductive system of an organism. Endosulfan is an organochlorine cyclodiene pesticide used widely for controlling agricultural pests. It has been shown to induce reproductive dysfunctions such as sperm abnormalities, reduced intracellular spermatid count in exposed organisms. Germ cells being the progenitor cells for male gametes and Sertoli cells as their nourishing cells, we examined whether endosulfan induces cell death in Sertoli-germ cells of male rats. Sertoli-germ cells, isolated from 28 d old male Wistar rats, were exposed to endosulfan (2.0, 20.0 and 40.0 ?g mL(-1)) for 24-72 h. Cytotoxicity, endosulfan concentration, reactive oxygen species (ROS) generation, oxidative stress parameters were measured in these cells in the absence or presence of endosulfan for the above mentioned exposure periods and subsequently, cell death endpoints were measured. We detected endosulfan in the exposed cells and demonstrated increased cell death in exposed Sertoli-germ cells as evidenced by a significant increase in annexin-V staining, depolarization of mitochondrial membrane, caspase-9 and -3 activities and BAD and PARP cleavage activities and DNA ladder formation along with non-significant increase in autophagic cell death. The study suggests that endosulfan can cause cell death in exposed Sertoli-germ cells due to higher oxidative damage with the activation of intrinsic cell death pathway which may eventually affect the production of quality gametes. PMID:24125708

Rastogi, Divya; Narayan, R; Saxena, D K; Chowdhuri, D Kar

2014-01-01

80

Concomitant Sertoli and Leydig Cell Tumor of the Testis: A Case Report  

PubMed Central

A rare intratubular gonadal stromal tumor was present in the testis of a 45-year-old man who was admitted to our hospital with the chief complaint of gradual enlargement of the left testis. Tumoral markers were negative and no extension was observed. The tumor comprised an intratubular mixture of two types of tumor cells with intercellular junctions: the predominant tumor cells were consistent with a Sertoli cell origin and cells comprising the minor population consistent with a Leydig cell origin. The patient is disease free after 6-month follow-up. The case is considered to be a testicular mixed tubular Sertoli-Leydig cell tumor. It highlights a rare type of primary tumor of the testis that features a good prognosis.

Tazi, Mohammed Fadl; Ahallal, Youness; Khallouk, Abdelhak; Elfatemi, Hinde; Bendahou, Mohcine; Tazi, Elmehdi; El Fassi, Mohammed Jamal; Farih, Moulay Hassan

2011-01-01

81

Rat testicular germ cells and sertoli cells release different types of bioactive transforming growth factor beta in vitro  

PubMed Central

Several in vivo studies have reported the presence of immunoreactive transforming growth factor-?'s (TGF-?'s) in testicular cells at defined stages of their differentiation. The most pronounced changes in TGF-?1 and TGF-?2 immunoreactivity occurred during spermatogenesis. In the present study we have investigated whether germ cells and Sertoli cells are able to secrete bioactive TGF-?'s in vitro, using the CCl64 mink lung epithelial cell line as bioassay for the measurement of TGF-?. In cellular lysates, TGF-? bioactivity was only observed following heat-treatment, indicating that within these cells TGF-? is present in a latent form. To our surprise, active TGF-? could be detected in the culture supernatant of germ cells and Sertoli cells without prior heat-treatment. This suggests that these cells not only produce and release TGF-? in a latent form, but that they also release a factor which can convert latent TGF-? into its active form. Following heat-activation of these culture supernatant's, total TGF-? bioactivity increased 6- to 9-fold. Spermatocytes are the cell type that releases most bioactive TGF-? during a 24 h culture period, although round and elongated spermatids and Sertoli cells also secrete significant amounts of TGF-?. The biological activity of TGF-? could be inhibited by neutralizing antibodies against TGF-?1 (spermatocytes and round spermatids) and TGF-?2 (round and elongating spermatids). TGF-? activity in the Sertoli cell culture supernatant was inhibited slightly by either the TGF-?1 and TGF-?2 neutralizing antibody. These in vitro data suggest that germ cells and Sertoli cells release latent TGF-?'s. Following secretion, the TGF-?'s are converted to a biological active form that can interact with specific TGF-? receptors. These results strengthen the hypothesis that TGF-?'s may play a physiological role in germ cell proliferation/differentiation and Sertoli cell function.

Haagmans, Bart L; Hoogerbrugge, Jos W; Themmen, Axel PN; Teerds, Katja J

2003-01-01

82

Environmentally Induced Epigenetic Transgenerational Inheritance of Altered Sertoli Cell Transcriptome and Epigenome: Molecular Etiology of Male Infertility  

PubMed Central

Environmental toxicants have been shown to induce the epigenetic transgenerational inheritance of adult onset disease, including testis disease and male infertility. The current study was designed to determine the impact of an altered sperm epigenome on the subsequent development of an adult somatic cell (Sertoli cell) that influences the onset of a specific disease (male infertility). A gestating female rat (F0 generation) was exposed to the agriculture fungicide vinclozolin during gonadal sex determination and then the subsequent F3 generation progeny used for the isolation of Sertoli cells and assessment of testis disease. As previously observed, enhanced spermatogenic cell apoptosis was observed. The Sertoli cells provide the physical and nutritional support for the spermatogenic cells. Over 400 genes were differentially expressed in the F3 generation control versus vinclozolin lineage Sertoli cells. A number of specific cellular pathways were identified to be transgenerationally altered. One of the key metabolic processes affected was pyruvate/lactate production that is directly linked to spermatogenic cell viability. The Sertoli cell epigenome was also altered with over 100 promoter differential DNA methylation regions (DMR) modified. The genomic features and overlap with the sperm DMR were investigated. Observations demonstrate that the transgenerational sperm epigenetic alterations subsequently alters the development of a specific somatic cell (Sertoli cell) epigenome and transcriptome that correlates with adult onset disease (male infertility). The environmentally induced epigenetic transgenerational inheritance of testis disease appears to be a component of the molecular etiology of male infertility.

Guerrero-Bosagna, Carlos; Savenkova, Marina; Haque, Md. Muksitul; Nilsson, Eric; Skinner, Michael K.

2013-01-01

83

Expression of a novel factor, short-type PB-cadherin, in Sertoli cells and spermatogenic stem cells of the neonatal rat testis  

Microsoft Academic Search

In the rodent testis, contact-mediated interactions between gonocytes, or neonatal stem cells, and Sertoli cells are critical for development. Previously, we showed that the neural cell adhesion molecule (NCAM) serves as a Sertoli cell-gonocyte attachment factor in neonates. Its expression decreases dramatically by 1 week of age and eventually disappears in vivo, and appears to be down- regulated by thyroid

J Wu; W F Jester Jr; A L Laslett; A Meinhardt; J M Orth

2003-01-01

84

Notch signaling in Sertoli cells regulates cyclical gene expression of Hes1 but is dispensable for mouse spermatogenesis.  

PubMed

Mammalian spermatogenesis is a highly regulated system dedicated to the continuous production of spermatozoa from spermatogonial stem cells, and the process largely depends on microenvironments created by Sertoli cells, unique somatic cells that reside within a seminiferous tubule. Spermatogenesis progresses with a cyclical program known as the "seminiferous epithelial cycle," which is accompanied with cyclical gene expression changes in Sertoli cells. However, it is unclear how the cyclicity in Sertoli cells is regulated. Here, we report that Notch signaling, which is known to play an important role for germ cell development in Drosophila and Caenorhabditis elegans, is cyclically activated in Sertoli cells and regulates stage-dependent gene expression of Hes1. To elucidate the regulatory mechanism of stage-dependent Hes1 expression and the role of Notch signaling in mouse spermatogenesis, we inactivated Notch signaling in Sertoli cells by deleting protein O-fucosyltransferase 1 (Pofut1), using the cre-loxP system, and found that stage-dependent Hes1 expression was dependent on the activation of Notch signaling. Unexpectedly, however, spermatogenesis proceeded normally. Our results thus indicate that Notch signaling regulates cyclical gene expression in Sertoli cells but is dispensable for mouse spermatogenesis. This highlights the evolutionary divergences in regulation of germ cell development. PMID:22037762

Hasegawa, Kazuteru; Okamura, Yoshiaki; Saga, Yumiko

2012-01-01

85

RiboTag Analysis of Actively Translated mRNAs in Sertoli and Leydig Cells In Vivo  

PubMed Central

Male spermatogenesis is a complex biological process that is regulated by hormonal signals from the hypothalamus (GnRH), the pituitary gonadotropins (LH and FSH) and the testis (androgens, inhibin). The two key somatic cell types of the testis, Leydig and Sertoli cells, respond to gonadotropins and androgens and regulate the development and maturation of fertilization competent spermatozoa. Although progress has been made in the identification of specific transcripts that are translated in Sertoli and Leydig cells and their response to hormones, efforts to expand these studies have been restricted by technical hurdles. In order to address this problem we have applied an in vivo ribosome tagging strategy (RiboTag) that allows a detailed and physiologically relevant characterization of the “translatome” (polysome-associated mRNAs) of Leydig or Sertoli cells in vivo. Our analysis identified all previously characterized Leydig and Sertoli cell-specific markers and identified in a comprehensive manner novel markers of Leydig and Sertoli cells; the translational response of these two cell types to gonadotropins or testosterone was also investigated. Modulation of a small subset of Sertoli cell genes occurred after FSH and testosterone stimulation. However, Leydig cells responded robustly to gonadotropin deprivation and LH restoration with acute changes in polysome-associated mRNAs. These studies identified the transcription factors that are induced by LH stimulation, uncovered novel potential regulators of LH signaling and steroidogenesis, and demonstrate the effects of LH on the translational machinery in vivo in the Leydig cell.

Sanz, Elisenda; Evanoff, Ryan; Quintana, Albert; Evans, Elizabeth; Miller, Jeremy A.; Ko, Chemyong; Amieux, Paul S.; Griswold, Michael D.; McKnight, G. Stanley

2013-01-01

86

Bilateral Sertoli-Leydig Cell Tumor in a Primigravida: A Rare Case  

PubMed Central

We present a unique case of incidentally discovered bilateral Sertoli Leydig cell tumour in a primigravida who displayed no features of virilization. The apha fetoprotein levels were elevated. Magnetic resonance imaging was suggestive of ovarian tumors, possibly germ cell tumor. Bilateral salpingo-oophorectomy was performed and histopathology showed features of Sertoli Leydig cell tumor with intermediate to poor differentiation. Immunohistochemistry was positive for calretinin and inhibin, while cytokeratin was negative. Four courses of bleomycin-, etoposide- and cisplatin-based chemotherapy regimen was started, but the patient aborted while receiving the second cycle of chemotherapy. She received the remaining two cycles of chemotherapy and is now on close follow up with monitoring of serum inhibin levels to detect any tumor recurrence. Bilateral Sertloli Leydig cell tumor has not been reported previously in a pregnant female. The aim of this article is to describe the clinical, radiological and pathological features and management of this rare entity.

Tyagi, Ruchita; Agrawal, Parimal; Nijhawan, Raje; Prasad, GRV

2014-01-01

87

Multiple Neoplasia in a Captive Jungle Cat ('Felis chaus)-Thyroid Adenocarcinoma, Gastric Adenocarcinoma, Renal Adenoma, and Sertoli Cell Tumor.  

National Technical Information Service (NTIS)

A 23-year-old captive male jungle cat (Felis chaus) was euthanatized because of declining health and advanced age. The following neoplasms were identified at necropsy: adenocarcinoma of the thyroid gland, adenocarcinoma of the stomach, Sertoli cell tumor,...

F. M. Garner J. W. Sagartz R. M. Sauer

1972-01-01

88

Nuclear localization of the actin regulatory protein Palladin in sertoli cells.  

PubMed

In the testis, F-actin structures are involved in spermatid nuclear remodeling and cytoplasm reduction, maintenance of the blood-testis barrier, support of the spermatogonial stem cell niche, and release of spermatids into the tubular lumen. To gain a better understanding of actin regulation in Sertoli-germ cell interactions, we investigated the expression of the Palladin (Palld) gene, which encodes a widely expressed phosphoprotein that localizes to actin-rich cytoplasmic structures, including focal adhesions, cell-cell junctions, podosomes, and stress fibers, and serves as a molecular scaffold to bundle actin fibers. In germ cells, PALLD was concentrated along the tubulin- and F-actin-containing cytoplasmic manchette that forms adjacent to the elongating spermatid nucleus during spermiogenesis. To our surprise, PALLD relocated from the cytoplasm to the nucleus of Sertoli cells in the juvenile testis, coincident with the onset of puberty, and this localization was maintained in the adult. We provide evidence that the 140 kDa isoform of PALLD predominates in Sertoli cells, and that it is apparently cleaved, with the C-terminus localizing to the nucleus while the N-terminus remains cytoplasmic. We investigated the nuclear localization of the C-terminus of PALLD and found that it is regulated by a putative nuclear export signal. These results provide the foundation for future work employing Sertoli cell- and spermatid-specific Palld-knockout mice to study diverse roles of PALLD as both a nuclear-actin regulatory protein and as a potential regulator of manchette formation during spermatogenesis. PMID:23559268

Niedenberger, Bryan A; Chappell, Vesna K; Kaye, Evelyn P; Renegar, Randall H; Geyer, Christopher B

2013-05-01

89

AKAP9 Is Essential for Spermatogenesis and Sertoli Cell Maturation in Mice  

PubMed Central

Mammalian male fertility relies on complex inter- and intracellular signaling during spermatogenesis. Here we describe three alleles of the widely expressed A-kinase anchoring protein 9 (Akap9) gene, all of which cause gametogenic failure and infertility in the absence of marked somatic phenotypes. Akap9 disruption does not affect spindle nucleation or progression of prophase I of meiosis but does inhibit maturation of Sertoli cells, which continue to express the immaturity markers anti-Mullerian hormone and thyroid hormone receptor alpha in adults and fail to express the maturation marker p27Kip1. Furthermore, gap and tight junctions essential for blood–testis barrier (BTB) organization are disrupted. Connexin43 (Cx43) and zona occludens-1 are improperly localized in Akap9 mutant testes, and Cx43 fails to compartmentalize germ cells near the BTB. These results identify and support a novel reproductive tissue-specific role for Akap9 in the coordinated regulation of Sertoli cells in the testis.

Schimenti, Kerry J.; Feuer, Sky K.; Griffin, Laurie B.; Graham, Nancy R.; Bovet, Claire A.; Hartford, Suzanne; Pendola, Janice; Lessard, Carl; Schimenti, John C.; Ward, Jeremy O.

2013-01-01

90

Distribution of vimentin-type intermediate filaments in Sertoli cells of the human testis, normal and pathologic  

Microsoft Academic Search

Summary  The presence, distribution and spatial arrangement of vimentin-type intermediate filaments in Sertoli cells from human testis\\u000a biopsies, were studied in semithin and ultrathin sections using a polyclonal rabbit antiserum. At the ultrastructural level,\\u000a vimentin immunoreactivity was seen concentrated around the nuclei, along fibrillary material within the cytoplasm and at the\\u000a ectoplasmic specializations of the Sertoli cell junctions, as well as

Gerhard Aumiiller; Manfred Steinbriick; Walter Krause; Hans-Joachim Wagner

1988-01-01

91

Immunoactive Inhibin as a Marker of Sertoli Cell Function following Cytotoxic Damage to the Human Testis  

Microsoft Academic Search

Sertoli and Leydig cell functions were evaluated in men with testicular damage due either to cytotoxic chemotherapy (CCT) or radiotherapy (XRT). Serum immunoactive inhibin, follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone concentrations were measured in 15 men (19–50 years) who had received 6–10 courses of combination CCT (mustine, vinblastine, procarbazine and prednisolone) for Hodgkin’s disease 1–8 years earlier and

A. Tsatsoulis; S. M. Shalet; I. D. Morris; D. M. de Kretser

1990-01-01

92

Correlative morphology and endocrinology of Sertoli cells in hamster testes in active and inactive states of spermatogenesis.  

PubMed

The seasonally breeding golden (Syrian) hamster, which exhibits photoperiod-dependent transitions between active and inactive states of spermatogenesis, was used as a model to study Sertoli cell structure in the two extreme phases of gonadal activity. The structural parameters of the Sertoli cell and its subcellular organelles were assessed using accepted stereological procedures during active and inactive states of spermatogenesis, and the results correlated with a battery of endocrine parameters obtained from the same animals. Short photoperiod-induced testicular involution was associated with a significant decrease in virtually all morphological parameters of the Sertoli cell, including a dramatic decrease in the volumes and surface areas of the Sertoli cells and their major subcellular organelles. Sertoli cell size and surface area were significantly and positively correlated with the testicular weight, volume of the seminiferous tubule, tubular lumena, tubule diameter, and germ cell numbers. Similar correlations were recorded between the number of germ cells and nearly all subcellular parameters of the Sertoli cell. Only those structural elements that are related to degredative processes (lysosomes and lipid) did not show significant volumetric differences between gonadally active and inactive animals. The observed changes in the structural parameters of the Sertoli cells were significantly correlated with the reduction in plasma levels of FSH, LH, and testosterone and intratesticular levels of testosterone. Exposure of hamsters to a short photoperiod was also associated with an increase in concentration (femtomoles per mg protein), but a decrease in the total content (femtomoles per testis) of testicular FSH receptors. The dissociation of changes in the content and concentration of FSH receptors appears to be related to changes in basal compartment plasma membrane surface areas of the Sertoli cells during testicular regression. The striking changes in Sertoli cell morphology between active and inactive states of spermatogenesis are structural manifestations of alterations in the function of these cells in response to the concomitant endocrine changes in the testis and indicate a virtual shut-down of Sertoli cell function during short photoperiod-induced testicular regression. PMID:2791968

Hikim, A P; Amador, A G; Klemcke, H G; Bartke, A; Russell, L D

1989-10-01

93

Retinol and retinoic acid increase MMP-2 activity by different pathways in cultured Sertoli cells.  

PubMed

Diseases such as atherosclerosis, arthritis and cancer have been related with imbalance in ROS production and failures in regulation of the MMPs. Authors suggested a relationship between MPP activity and ROS. Our research group has demonstrated that retinol 7 microM induced changes in Sertoli cell metabolism linking retinol treatment and oxidative stress. We verified MMP activity in Sertoli cells treated with vitamin A using gelatin zymography. We found that retinol (7 microM) and retinoic acid (1 nM) induced MMP-2 activity in Sertoli cells. Antioxidants reversed retinol-induced but not retinoic acid-induced MMP-2 activity. Moreover, retinol but not retinoic acid increased ROS production quantified by DCFH-DA oxidation. We found that retinol and retinoic acid induced ERK1/2 phosphorylation, but only retinol-increased MMP-2 activity was inhibited by UO126, an ERK1/2 phosphorylation inhibitor. Our findings suggested that retinol-induced MMP-2 activity, but not retinoic acid-induced MMP-2 activity, was related to ERK1/2 phosphorylation and ROS production. PMID:18075836

Dalmolin, Rodrigo J S; Zanotto-Filho, Alfeu; De Oliveira, Ramatis B; Duarte, Roxane F; Pasquali, Matheus A B; Moreira, José C F

2007-12-01

94

Low density lipoprotein receptor-related protein-1 expression in the testis: regulated expression in Sertoli cells.  

PubMed

The low density lipoprotein receptor-related protein (LRP-1) is a multiligand receptor capable of mediating endocytosis of a wide array of ligands that relate to both lipoprotein metabolism and proteinase regulation. Many of its ligands are proteinases, proteinase-inhibitor complexes, and lipoproteins known to be contained within the luminal fluid of the seminiferous tubules or in the interstitial spaces of the testis. Immunocytochemical analysis was performed to characterize the pattern of expression of LRP-1 in cells of the rat testis. Immunoperoxidase staining localized LRP-1 to the cytoplasm of Sertoli cells. The distribution and intensity of the Sertoli cell staining was found to vary according to the stages of the cycle of the seminiferous epithelium. Staining was weak in the basal cytoplasm of Sertoli cells during stages II-VIII and strong and granular in the supranuclear cytoplasm during stages XII-XIV and stage I of the cycle. Immunogold labeling showed gold particles associated with the basal and adluminal plasma membranes, with endocytic vesicles, and with endosome membranes. Labeling was also observed on the plasma membrane and membranes of the endocytic apparatus in macrophages and Leydig cells in the interstitial space. Infusion of 125I-Labeled LRP-1 antibody into seminiferous tubules followed by radioautography showed silver grains overlaying the ad-luminal plasma membrane of Sertoli cells at time 0 and in endocytic vesicles and endosomes in the supranuclear region of Sertoli cells 10-minutes postinjection. When the 125I-Labeled LRP-1 antibody was injected into the interstitial space, silver grains overlayed the basal plasma membrane and coated endocytic pits of Sertoli cells at time 0 and, at 10 minutes, the grains labeled endosomes located in the basal pole of Sertoli cells. 125I-Labeled LRP-1 antibody also labeled the plasma membrane and the endocytic apparatus of macrophages and Leydig cells. The absence of immunogold labeling and radioautographic silver grains within lysosomes of Sertoli cells, Leydig cells, and macrophages suggests that internalized LRP-1 is recycled back to the cell surface. The presence of LRP-1 in the endocytic compartment of these cells is consistent with its functioning in the clearance of proteases involved in seminiferous tubule remodeling and/ or the uptake of cholesterol-bound lipoproteins necessary for the biosynthesis of testosterone. In conclusion, the results of these studies demonstrated for the first time the presence of LRP-1 receptor in the endocytic compartments of Sertoli cells and interstitial cells of the rat testis. PMID:9283953

Igdoura, S A; Argraves, W S; Morales, C R

1997-01-01

95

miRNA regulation of cytotoxic effects in mouse Sertoli cells exposed to nonylphenol  

PubMed Central

Background It is known that some environmental chemicals affect the human endocrine system. The harmful effects of endocrine disrupting chemical (EDC) nonylphenol (NP) have been studied since the 1980s. It is known that NP adversely affects physiological functions by mimicking the natural hormone 17 beta-estradiol. In the present study, we analyzed the expression of miRNAs and their target genes in mouse Sertoli TM4 cells to better understand the regulatory roles of miRNAs on Sertoli cells after NP exposure. Methods Mouse TM4 Sertoli cells were treated with NP for 3 or 24 h, and global gene and miRNA expression were analyzed using Agilent mouse whole genome and mouse miRNA v13 arrays. Results We identified genes that were > 2-fold differentially expressed in NP-treated cells and control cells (P < 0.05) and analyzed their functions through Gene Ontology analysis. We also identified miRNAs that were differentially expressed in NP-treated and control cells. Of the 186 miRNAs the expression of which differed between NP-treated and control cells, 59 and 147 miRNAs exhibited 1.3-fold increased or decreased expression at 3 and 24 h, respectively. Network analysis of deregulated miRNAs suggested that Ppara may regulate the expression of certain miRNAs, including miR-378, miR-125a-3p miR-20a, miR-203, and miR-101a, after exposure to NP. Additionally, comprehensive analysis of predicted target genes for miRNAs showed that the expression of genes with roles in cell proliferation, the cell cycle, and cell death were regulated by miRNA in NP-treated TM4 cells. Levels of expression of the miRNAs miR-135a* and miR-199a-5p were validated by qRT-PCR. Finally, miR-135a* target gene analysis suggests that the generation of reactive oxygen species (ROS) following exposure to NP exposure may be mediated by miR-135a* through regulation of the Wnt/beta-catenin signaling pathway. Conclusions Collectively, these data help to determine NP's actions on mouse TM4 Sertoli cells and increase our understanding of the molecular mechanisms underlying the adverse effects of xenoestrogens on the reproductive system.

2011-01-01

96

Morphological changes of Sertoli cells during the male reproductive cycle of the teleost Piaractus mesopotamicus (Holmberg, 1887).  

PubMed

An investigation of the histological and ultrastructural changes of Sertoli cells during the male reproductive cycle in Piaractus mesopotamicus was made. The results showed that the Sertoli cell development is closely related with germ cell maturation. Therefore, these cells may have some role in germ cell maturation during the reproductive cycle of this species, whether in forming a tissue framework for the developing spermatogenic cysts, aiding in testes reorganization for a new reproductive cycle, in addition to other possible functions discussed in the text. PMID:16097726

Cruz-Landim, C; Abdalla, F C; Cruz-Höflinq, M A

2005-05-01

97

The Death of Sertoli Cells and the Capacity to Phagocytize Elongated Spermatids During Testicular Regression due to Short Photoperiod in Syrian Hamster (Mesocricetus auratus).  

PubMed

In the Syrian hamster (Mesocricetus auratus), an animal that displays testicular regression due to short photoperiod, germ cells are removed by apoptosis during this process and the apoptotic remains are phagocytized by Sertoli cells. The aim of this work was to investigate morphologically whether the testicular regression process due to short photoperiod leads to the apoptosis of Sertoli cells, and whether, during testicular regression, the elongated spermatids are eliminated through phagocytosis by Sertoli cells. To this end, we studied testis sections during testicular regression in Syrian hamster subjected to short photoperiod by means of several morphological techniques using conventional light microscopy (hematoxylin and eosin [H&E], semi-thin section vimentin, immunohistochemistry, SBA lectin, and TUNEL staining), fluorescence microscopy, and transmission electron microscopy (TEM). H&E and semi-thin sections identified Sertoli cells with a degenerated morphology. Greater portion of Sertoli cells that were positive for TUNEL staining were observed especially during the mild regression (MR) and strong regression (SR) phases. In addition, TEM identified the characteristic apoptotic changes in the nucleus and cytoplasm of Sertoli cells. Moreover, during testicular regression and using light microscopy, some elongated spermatids were seen in basal position next to the Sertoli cell nucleus. This Sertoli phagocytic activity was higher in MR and SR phases. TEM confirmed this to be the result of the phagocytic activity of Sertoli cells. In conclusion, during testicular regression in Syrian hamster due to short photoperiod, when germ cells are known to be lost through apoptosis, there is morphological evidences that Sertoli cells are also lost through apoptosis, while some elongated spermatids are phagocytized and eliminated by the Sertoli cells. PMID:24719257

Seco-Rovira, Vicente; Beltrán-Frutos, Esther; Ferrer, Concepción; Sáez, Francisco José; Madrid, Juan Francisco; Pastor, Luis Miguel

2014-05-01

98

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced cytotoxicity accompanied by oxidative stress in rat Sertoli cells: Possible role of mitochondrial fractions of Sertoli cells  

SciTech Connect

TCDD, as an endocrine disruptor, is known to impair testicular functions and fertility. To elucidate the mechanism(s) underlying the testicular effects of TCDD, the potential toxicity of TCDD on Sertoli cells was investigated. Furthermore, the study aims to delineate whether mitochondrial fractions of Sertoli cells are involved in mediating the testicular effects of TCDD. Adult rat Sertoli cells were incubated with (5, 10 or 15 nM) of TCDD for 6, 12 or 24 h. Cell viability, lactate and LDH leakage into media along with lipid peroxidation, ROS generation, SOD, CAT, GPx, GR, {gamma}-GT and {beta}-glucuronidase activities, GSH content and {Delta}{psi}{sub m} were measured. Superoxide anion production, COX and cardiolipin content were measured in mitochondrial fractions. Cell viability was significantly decreased while lactate and LDH leakage into media were increased. ROS generation along with lipid peroxidation was also increased. SOD, CAT, GPx, GR activities and GSH content were significantly decreased. {gamma}-GT and {beta}-glucuronidase activities were also decreased. Superoxide anion production was increased while COX activity and cardiolipin content were decreased in mitochondrial fractions. Moreover, the {Delta}{psi}{sub m} was significantly decreased as measured in Sertoli cells. In conclusion, TCDD impairs Sertoli cell functions and this effect is, at least in part, attributed to oxidative stress. We have also found that TCDD increases mitochondrial superoxide anion production and decreases {Delta}{psi}{sub m}, COX activity and mitochondrial cardiolipin content. Our findings suggest that mitochondria may play an important role in ROS production, leading to the TCDD-induced oxidative stress response and resulting toxicological consequences in rat Sertoli cells.

Aly, Hamdy A.A., E-mail: hamdyaali@yahoo.com [Department of Pharmacology and Toxicology, Faculty of Pharmacy, King Abdulaziz University, Jeddah (Saudi Arabia); Khafagy, Rasha M. [Physics Department, Girls College for Arts, Science and Education, Ain Shams University, Cairo (Egypt)

2011-05-01

99

1?,25(OH) 2Vitamin D 3 stimulation of secretion via chloride channel activation in Sertoli cells  

Microsoft Academic Search

Sertoli cell secretory activities are highly dependent on ion channel functions and critical to spermatogenesis. The steroid hormone 1?,25(OH)2-vitamin D3 (1,25(OH)2-D3) stimulates exocytosis in different cell systems by activating a nongenotropic vitamin D receptor (VDR). Here, we described 1,25(OH)2-D3 stimulation of secretion via Cl? channel activation in the mouse immature Sertoli cell line TM4. 1,25(OH)2-D3 potentiation of chloride currents was

Danusa Menegaz; Antonio Barrientos-Duran; Andrew Kline; Fatima R. M. B. Silva; Anthony W. Norman; Mathew T. Mizwicki; Laura P. Zanello

2010-01-01

100

Sertoli Leydig cell ovarian tumour and gastric polyps as presenting features of Peutz-Jeghers syndrome.  

PubMed

We report a case of Peutz-Jeghers syndrome (PJS) in a 2-year old with precocious puberty secondary to a Sertoli-Leydig cell tumour. Family history of PJS and other neoplasms were discovered. The tumour was excised and the STK11 gene deletion identified in both patient and father. Screening revealed hamartomatous gastric polyps, which were removed. Current recommendations for screening of children with PJS begin at age 8 years, based on reported occurrence of complications 1. This report illustrates the importance of considering early screening, along with close clinical review and patient/parent education, for detection of life threatening neoplasms and complications. PMID:20310004

Howell, Lisa; Bader, Abdulgader; Mullassery, Dhanya; Losty, Paul; Auth, Marcus; Kokai, George

2010-07-15

101

Follicle-stimulating hormone regulates both Sertoli cell and spermatogonial populations in the adult photoinhibited Djungarian hamster testis.  

PubMed

The hormones that regulate spermatogonial development are ill defined, in part due to lack of appropriate experimental models. The photoinhibited hamster model provides a rich source of spermatogonia, thus making it an ideal model to study their control. This study aimed to assess the effects of FSH, in the absence of testosterone, on the reinitiation of Sertoli cell and spermatogonial development in the photosensitive adult Djungarian hamster. Hamsters raised under long photoperiods (LD, 16L:8D) were exposed to short photoperiods (SD, 8L:16D) for 11 wk, leading to suppression of gonadotropins and regression of testicular function. Groups of 10 animals then received FSH alone or in combination with the antiandrogen, flutamide, for 7 days. Two control groups maintained either under long or short photoperiods were treated with vehicle. Sertoli and germ cell number were then determined using the optical disector (sic) stereological technique. The number of Sertoli cells, type A spermatogonia, type B spermatogonia/preleptotene spermatocytes, and leptotene/zygotene spermatocytes were suppressed in SD controls to 66%, 34%, 19%, and 10% (all P < 0.01) of long-day control values, respectively. Later germ cell types were not detected. FSH treatment, with or without flutamide, increased Sertoli cell number (P < 0.01) to normal long-day values. Similarly, FSH treatment in the absence/presence of flutamide increased type A spermatogonia, type B spermatogonia/preleptotene spermatocytes, and leptotene/zygotene spermatocytes to approximately 85%, 69%, and 80% (all P < 0.01) of long-day controls, respectively. Our data demonstrate that the reinitiation of spermatogonial maturation in this model is dependent on FSH in the presence of an antiandrogen. Surprisingly, the adult Sertoli cell population in this model is also hormone dependent. This naturally occurring model provides a unique opportunity to understand the mechanisms (apoptotic and/or proliferative) by which FSH regulates Sertoli and germ cell development in the adult animal. PMID:15659702

Meachem, Sarah J; Stanton, Peter G; Schlatt, Stefan

2005-05-01

102

Sertoli-sertoli junctions and Sertoli-spermatid junctions after efferent ductule ligation and lanthanum treatment.  

PubMed

Seminiferous tubules, partially dilated by ligation of the efferent ductules, were examined after treatment with lanthanum. Lanthanum penetrated the intercellular spaces of the seminiferous epithelium, but only to the level of the Sertoli-Sertoli junctions. Further penetration from the interstitial surface of the tubule was restricted by membrane fusions (tight junctions) at the junctional complex. Lanthanum also penetrated the epithelium from the luminal surface permeating the adluminal intercellular spaces, including the site of the Sertoli-spermatid junction. The lanthanum occupying the Sertoli-spermatid junctional site appeared as a slightly narrower electron-opaque zone than that found in the non-specialized intercellular areas. The findings clearly reveal that only the Sertoli-Sertoli junctional site forms a restrictive barrier. In contrast to the specializations of plasma membrane which form the tight junction, the associated filaments and cisterna of endoplasmic reticulum may be components more directly related to maintaining and regulating cell adhesion. PMID:842475

Ross, M H

1977-01-01

103

Retinoic acid induces Sertoli cell paracrine signals for spermatogonia differentiation but cell autonomously drives spermatocyte meiosis  

PubMed Central

Direct evidence for a role of endogenous retinoic acid (RA), the active metabolite of vitamin A in the initial differentiation and meiotic entry of spermatogonia, and thus in the initiation of spermatogenesis is still lacking. RA is synthesized by dedicated enzymes, the retinaldehyde dehydrogenases (RALDH), and binds to and activates nuclear RA receptors (RARA, RARB, and RARG) either within the RA-synthesizing cells or in the neighboring cells. In the present study, we have used a combination of somatic genetic ablations and pharmacological approaches in vivo to show that during the first, prepubertal, spermatogenic cycle (i) RALDH-dependent synthesis of RA by Sertoli cells (SC), the supporting cells of the germ cell (GC) lineage, is indispensable to initiate differentiation of A aligned into A1 spermatogonia; (ii) RARA in SC mediates the effects of RA, possibly through activating Mafb expression, a gene whose Drosophila homolog is mandatory to GC differentiation; (iii) RA synthesized by premeiotic spermatocytes cell autonomously induces meiotic initiation through controlling the RAR-dependent expression of Stra8. Furthermore, we show that RA of SC origin is no longer necessary for the subsequent spermatogenic cycles but essential to spermiation. Altogether, our data establish that the effects of RA in vivo on spermatogonia differentiation are indirect, via SC, but direct on meiotic initiation in spermatocytes, supporting thereby the notion that, contrary to the situation in the female, RA is necessary to induce meiosis in the male.

Raverdeau, Mathilde; Gely-Pernot, Aurore; Feret, Betty; Dennefeld, Christine; Benoit, Gerard; Davidson, Irwin; Chambon, Pierre; Mark, Manuel; Ghyselinck, Norbert B.

2012-01-01

104

Retinoic acid induces Sertoli cell paracrine signals for spermatogonia differentiation but cell autonomously drives spermatocyte meiosis.  

PubMed

Direct evidence for a role of endogenous retinoic acid (RA), the active metabolite of vitamin A in the initial differentiation and meiotic entry of spermatogonia, and thus in the initiation of spermatogenesis is still lacking. RA is synthesized by dedicated enzymes, the retinaldehyde dehydrogenases (RALDH), and binds to and activates nuclear RA receptors (RARA, RARB, and RARG) either within the RA-synthesizing cells or in the neighboring cells. In the present study, we have used a combination of somatic genetic ablations and pharmacological approaches in vivo to show that during the first, prepubertal, spermatogenic cycle (i) RALDH-dependent synthesis of RA by Sertoli cells (SC), the supporting cells of the germ cell (GC) lineage, is indispensable to initiate differentiation of A aligned into A1 spermatogonia; (ii) RARA in SC mediates the effects of RA, possibly through activating Mafb expression, a gene whose Drosophila homolog is mandatory to GC differentiation; (iii) RA synthesized by premeiotic spermatocytes cell autonomously induces meiotic initiation through controlling the RAR-dependent expression of Stra8. Furthermore, we show that RA of SC origin is no longer necessary for the subsequent spermatogenic cycles but essential to spermiation. Altogether, our data establish that the effects of RA in vivo on spermatogonia differentiation are indirect, via SC, but direct on meiotic initiation in spermatocytes, supporting thereby the notion that, contrary to the situation in the female, RA is necessary to induce meiosis in the male. PMID:23012458

Raverdeau, Mathilde; Gely-Pernot, Aurore; Féret, Betty; Dennefeld, Christine; Benoit, Gérard; Davidson, Irwin; Chambon, Pierre; Mark, Manuel; Ghyselinck, Norbert B

2012-10-01

105

Can electrons travel through actin microfilaments and generate oxidative stress in retinol treated Sertoli cell?  

PubMed

In early reports our research group has demonstrated that 7 microM retinol (vitamin A) treatment leads to many changes in Sertoli cell metabolism, such as up-regulation of antioxidant enzyme activities, increase in damage to biomolecules, abnormal cellular division, pre-neoplasic transformation, and cytoskeleton conformational changes. These effects were observed to be dependent on the production of reactive oxygen species (ROS), suggesting extra-nuclear (non-genomic) effects of retinol metabolism. Besides 7 microM retinol treatment causing oxidative stress, we have demonstrated that changes observed in cytoskeleton of Sertoli cells under these conditions were protective, and seem to be an adaptive phenomenon against a pro-oxidant environment resulting from retinol treatment. We have hypothesized that the cytoskeleton can conduct electrons through actin microfilaments, which would be a natural process necessary for cell homeostasis. In the present study we demonstrate results correlating retinol metabolism, actin architecture, mitochondria physiology and ROS, in order to demonstrate that the electron conduction through actin microfilaments might explain our results. We believe that electrons produced by retinol metabolism are dislocated through actin microfilaments to mitochondria, and are transferred to electron transport chain to produce water. When mitochondria capacity to receive electrons is overloaded, superoxide radical production is increased and the oxidative stress process starts. Our results suggested that actin cytoskeleton is essential to oxidative stress production induced by retinol treatment, and electrons conduction through actin microfilaments can be the key of this correlation. PMID:17203241

de Oliveira, Ramatis Birnfeld; de Bittencourt Pasquali, Matheus Augusto; Filho, Alfeu Zanotto; Dalmolin, Rodrigo Juliani Siqueira; Gelain, Daniel Pens; Gottfried, Carmem; Rodrigues, José Luiz; Klamt, Fábio; Moreira, José Cláudio Fonseca

2007-07-01

106

Retinoic acid signaling in Sertoli cells regulates organization of the blood-testis barrier through cyclical changes in gene expression.  

PubMed

Mammalian spermatogenesis contributes a constant production of large numbers of spermatozoa, which is achieved by a cyclically regulated program known as the seminiferous epithelial cycle. Sertoli cells, functionally unique somatic cells, create a microenvironment to support the continuous differentiation of germ cells especially through the formation of a blood-testis barrier (BTB). The BTB is essential for maintaining homeostasis in seminiferous tubules and opens transiently at stages VII-VIII to ensure constant differentiation of spermatogenic cells. However, it is poorly understood how the dynamic organization of BTB is regulated. In our current study, we find that the overexpression of a dominant-negative form of RAR? (dnRAR?) in Sertoli cells disrupts the BTB at stages VII-XII and causes the large-scale apoptosis of differentiating germ cells. These abnormal events are found to be associated with cyclical gene expression changes in Sertoli cells, which can be represented by abnormal activation and repression of genes showing peaks of expression during stages I-VI and VII-XII, respectively. We find that one such gene, Ocln, encoding a tight junction component, partly contributes to the BTB disruption caused by dnRAR?. Taken together, our data suggest that the cyclical activation of RA signaling in Sertoli cells during stages VII-XII contributes to a periodic organization of the BTB through changes in stage-dependent gene expression. PMID:23095883

Hasegawa, Kazuteru; Saga, Yumiko

2012-12-01

107

Effect of sodium molybdate on the binding of androgen-receptor complexes to germ cell and Sertoli cell chromatin.  

PubMed

This study explores the effect of molybdate on androgen receptors in relation to chromatin binding. Two fractions of testicular and prostatic cytosol were obtained by ammonium sulfate precipitation at 15-37% (I) and 37-47% (II) saturation. Both fractions from either tissue exhibited specific binding of [3H]-androgens which was thermolabile. With the exception of testicular fraction II, all fractions bound to Sertoli cell and germ cell chromatin. Prostatic [3H]-androgen-receptor complexes isolated from cytosols in the presence of 10 mM sodium molybdate showed a decreased ability (only 20-40%) to bind to either Sertoli cell or germ cell chromatin. Similarly, testicular [3H]-androgen-receptor complexes isolated in the presence of molybdate bound less well (60-70%) to Sertoli cell chromatin. On the other hand, inclusion of molybdate subsequent to ammonium sulfate precipitation did not significantly alter the binding ability of the receptor complexes to either chromatin. Also, the presence of molybdate during the receptor-chromatin interaction did not significantly decrease the ability of either prostatic or testicular androgen-receptor complexes to bind to Sertoli cell chromatin. These results indicate that the addition of molybdate prior to ammonium sulfate precipitation may impede the activation of androgen-receptor complexes by salt, resulting in a decreased ability of the steroid-receptor complexes to bind to chromatin acceptor sites. The data also indicate that molybdate exerts its inhibitory effect directly on the nonactivated receptor complexes and not on the chromatin acceptor sites. PMID:7109600

Tsai, Y H; Steinberger, A

1982-08-01

108

Mouse Sertoli cells sustain de novo generation of regulatory T cells by triggering the notch pathway through soluble JAGGED1.  

PubMed

FOXP3(+) regulatory T cells (Tregs) are central to the maintenance of immunological homeostasis and tolerance. It has long been known that Sertoli cells are endowed with immune suppressive properties; however, the underlying mechanisms as well as the effective nature and role of soluble factors secreted by Sertoli cells have not been fully elucidated as yet. We hypothesized that conditioned medium from primary mouse Sertoli cells (SCCM) may be able and sufficient to induce Tregs. By culturing CD4(+)CD25(-)EGFP(-) T splenocytes purified from FOXP3-EGFP knock-in mice in SCCM, here we show, by flow cytometry and suppression assay, the conversion of peripheral CD4(+)FOXP3(-) T cells into functional CD4(+)FOXP3(+) Tregs. We also demonstrate that the Notch/Jagged1 axis is involved in regulating the de novo generation of Tregs although this process is transforming growth factor-beta1 (TGF-B) dependent. In particular, we identified by Western blot analysis a soluble form of JAGGED1 (JAG1) in SCCM that significantly influences the induction of Tregs, as demonstrated by performing the conversion assay in presence of a JAG1-specific neutralizing antibody. In addition, we show that SCCM modulates the Notch pathway in converted Tregs by triggering the recruitment of the Notch-specific transcription factor CSL/RBP-Jk to the Foxp3 promoter and by inducing the Notch target gene Hey1, as shown by chromatin immunoprecipitation assay and by real time-RT-PCR experiments, respectively. Overall, these results contribute to a better understanding of the molecular mechanisms involved in Sertoli cell-mediated immune tolerance and provide a novel approach to generate ex vivo functional Tregs for therapeutic purpose. PMID:24478388

Campese, Antonio Francesco; Grazioli, Paola; de Cesaris, Paola; Riccioli, Anna; Bellavia, Diana; Pelullo, Maria; Padula, Fabrizio; Noce, Claudia; Verkhovskaia, Sofia; Filippini, Antonio; Latella, Giovanni; Screpanti, Isabella; Ziparo, Elio; Starace, Donatella

2014-03-01

109

Involvement of a membrane skeletal protein, 4.1G, for Sertoli/germ cell interaction.  

PubMed

We previously reported that a membrane skeletal protein, 4.1G (also known as EPB41L2), is immunolocalized in mouse seminiferous tubules. In this study, the 4.1G immunolocalizaiton was precisely evaluated at various stages of the mouse seminiferous epithelial cycle with 'in vivo cryotechnique' and also with pre-embedding immunoelectron microscopy in testicular tissues whose ultrastructures were well preserved with glycerol treatment before cryosectioning. In addition, 4.1G-deficient mice were produced, and the morphology of their seminiferous tubules was also evaluated. The 4.1G immunolocalization was different among stages, indicating that it was not only along cell membranes of Sertoli cells, but also those of spermatogonia and early spermatocytes. To confirm the 4.1G immunolocalization in germ cells, in vitro culture of spermatogonial stem cells (SSCs) was used for immunocytochemistry and immunoblotting analysis. In the cultured SSCs, 4.1G was clearly expressed and immunolocalized along cell membranes, especially at mutual attaching regions. In testicular tissues, cell adhesion molecule-1 (CADM1), an intramembranous adhesion molecule, was colocalized on basal parts of the seminiferous tubules and immunoprecipitated with 4.1G in the tissue lysate. Interestingly, in the 4.1G-deficient mice, histological manifestation of the seminiferous tubules was not different from that in wild-type mice, and the CADM1 was also immunolocalized in the same pattern as that in the wild-type. Moreover, the 4.1G-deficient male mice were fertile. These results were probably due to functional redundancy of unknown membrane skeletal molecules in germ cells. Thus, a novel membrane skeletal protein, 4.1G, was found in germ cells, and considering its interaction with CADM family, it probably has roles in attachment of both Sertoli-germ and germ-germ cells. PMID:20200204

Terada, Nobuo; Ohno, Nobuhiko; Saitoh, Sei; Saitoh, Yurika; Komada, Masayuki; Kubota, Hiroshi; Ohno, Shinichi

2010-05-01

110

Vectorial (transcellular) transport of potassium (86Rb+) by cultured Sertoli cells.  

PubMed

Sertoli cells from rats aged 25 days were grown on Millipore filters (pore diameter 0.5 micron) for 7 days and were then used for determination of transport of 86Rb+ through the cells (base to apex); this procedure is referred to as measuring transcellular or vectorial transport. Sertoli cells were also used to measure apical efflux of 86Rb+ by loading the cells with the isotope to steady state and then incubating cells so that the apical surfaces were in contact with medium not containing 86Rb+, from which samples were taken. Basal efflux was measured in the same way except that the opposite surface of the cells was in contact with the medium. Cells grown on filters treated with collagen IV plus fibronectin showed transcellular transport of 86Rb+; t1/2 for equilibration across the cells was 9-12 min. The rate of transport was accelerated by addition of (Bu)2cAMP, forskolin, or FSH to the incubation medium. Half-maximal responses were seen with (Bu)2cAMP at 0.2 mM and with forskolin at 20 microM. Apical efflux (t1/2 9.8 +/- 2.1 min) was not influenced by the presence or absence of K+ in the medium nor by azide or (Bu)2cAMP. Basal efflux showed similar values for t1/2 in the presence of K+ (9.7 +/- 1.9 min) and values of 21.4 +/- 4.2 min in the absence of K+. Vectorial transport of 86Rb+ by these cells may account for the K+ gradient seen in the seminiferous tubule and appears to result from a basolateral potassium pump together with an apical membrane that is permeable to K+. PMID:2843358

Muffly, K E; Hall, P F

1988-10-01

111

Vectorial (transcellular) transport of potassium (/sup 86/Rb+) by cultured Sertoli cells  

SciTech Connect

Sertoli cells from rats aged 25 days were grown on Millipore filters (pore diameter 0.5 micron) for 7 days and were then used for determination of transport of 86Rb+ through the cells (base to apex); this procedure is referred to as measuring transcellular or vectorial transport. Sertoli cells were also used to measure apical efflux of 86Rb+ by loading the cells with the isotope to steady state and then incubating cells so that the apical surfaces were in contact with medium not containing 86Rb+, from which samples were taken. Basal efflux was measured in the same way except that the opposite surface of the cells was in contact with the medium. Cells grown on filters treated with collagen IV plus fibronectin showed transcellular transport of 86Rb+; t1/2 for equilibration across the cells was 9-12 min. The rate of transport was accelerated by addition of (Bu)2cAMP, forskolin, or FSH to the incubation medium. Half-maximal responses were seen with (Bu)2cAMP at 0.2 mM and with forskolin at 20 microM. Apical efflux (t1/2 9.8 +/- 2.1 min) was not influenced by the presence or absence of K+ in the medium nor by azide or (Bu)2cAMP. Basal efflux showed similar values for t1/2 in the presence of K+ (9.7 +/- 1.9 min) and values of 21.4 +/- 4.2 min in the absence of K+. Vectorial transport of 86Rb+ by these cells may account for the K+ gradient seen in the seminiferous tubule and appears to result from a basolateral potassium pump together with an apical membrane that is permeable to K+.

Muffly, K.E.; Hall, P.F.

1988-10-01

112

Morphometry and morphology of nucleus of the Sertoli and interstitial cells of the tambaqui Colossoma macropomum (Cuvier, 1881) (Pisces: Characidae) during the reproductive cycle.  

PubMed

This study allowed the characterization of the tambaqui Colossoma macropomum testes structural organization, emphasizing Sertoli and interstitial cells and analyzing morphometrically the Sertoli cell nucleus diameter and the interstitial tissue area during the reproductive cycle. Fragments of tambaqui testes were collected in the following reproductive cycle stages: immature, resting, maturation I and II, mature, and regression, and were histologically processed. The Sertoli cells were found at the periphery of the cysts of germinative lineage cells and the nuclei were shown to be smaller as these cells developed. The interstitial cells were better observed between the seminiferous lobules next to vessels in the interstitial tissue of maturing testes. PMID:12914420

Nakaghi, L S O; Mitsuiki, D; Santos, H S L; Pacheco, M R; Ganeco, L N

2003-02-01

113

Extracellular inosine participates in tumor necrosis factor-alpha induced nitric oxide production in cultured Sertoli cells.  

PubMed

Recent reports have described purinergic modulation of tumor necrosis factor-alpha (TNF-alpha) signaling in neutrophils and astrocytes. In Sertoli cells, both TNF-R1 and TNF-R2 TNF-alpha receptors are present and this cytokine modulates many functions of these cells related to the maintenance of spermatogenesis. Sertoli cells express distinct purinoreceptors and previous work has shown that these cells secrete extracellular nucleotides and their metabolites. In this work, we studied the possible role of extracellular purines in TNF-alpha signaling in cultured Sertoli cells. This cytokine increased inosine concentration from 30 min to 6 h, with no effect at 24 h. Both TNF-alpha and inosine increased nitrite accumulation and nitric oxide synthase activity. Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), an adenosine deaminase inhibitor, abolished the TNF-alpha induced inosine increase, nitrite accumulation and nitric oxide synthase activity. These results suggest that extracellular inosine acts as intermediary in TNF-alpha stimulated nitric oxide production in cultured Sertoli cells. PMID:16328964

de Souza, Luiz Fernando; Gelain, Daniel Pens; Jardim, Fernanda Rafaela; Ribeiro, Gisele Roncheti; Zim, Marcelo; Bernard, Elena Aida

2006-01-01

114

A light microscopic and morphometric analysis of the Sertoli cell during the spermatogenic cycle of the rat  

Microsoft Academic Search

Testes from 8 adult rats were perfusion-fixed with buffered glutaraldehyde and semi-thin sections of the seminiferous epithelium at all stages of the spermatogenic cycle were subjected to morphometric analysis at the light microscope level. Methods are presented to derive the volume occupied by the Sertoli cells within the total volume of the seminiferous epithelium at each stage of the cycle

Jeffrey B. Kerr

1988-01-01

115

The role of androgens in sertoli cell proliferation and functional maturation: studies in mice with total or Sertoli cell-selective ablation of the androgen receptor.  

PubMed

The role of androgens in the proliferation and maturation of Sertoli cells (SC) and the development of their capacity to support spermatogenesis remains poorly understood. We evaluated these functions in complete androgen receptor knockout (ARKO) and SC-selective androgen receptor knockout (SCARKO) mice. Compared with controls, ARKO mice exhibited a progressive reduction in SC number/testis, whereas SCARKOs showed minor changes, suggesting that androgen effects on SC number are not mediated via direct action on SCs. Immunoexpression of anti-Mullerian hormone (AMH), p27(kip1), GATA-1, and sulfated glycoprotein-2, which changes according to SC maturational status, occurred normally in ARKOs and SCARKOs. Functional capacity of SCs to support spermatogonia was similar in SCARKOs and controls, whereas ARKOs showed reduced capacity with age. SC capacity to support total germ cells revealed major deficits in ARKO and SCARKO adults, particularly with respect to postmeiotic germ cells. Using quantitative RT-PCR, the expression of SC markers was compared in d 50 testes. In ARKOs, expression of Pem, fatty acid binding protein, platelet-derived growth factor-A, and transferrin were all significantly reduced, whereas FSH receptor and AMH were increased. In SCARKOs, there were modest reductions in expression of cystatin-related gene highly expressed in testis and epididymis (cystatin-TE) and claudin-11, whereas expression of Pem, fatty acid binding protein, and platelet-derived growth factor-A was markedly reduced, highlighting these as potentially androgen-regulated SC genes that merit further study. In conclusion, androgen action is not required for maturation-dependent changes in immunoexpression of the SC markers AMH, p27(kip1), GATA-1, and sulfated glycoprotein-2 but is essential for expression of other SC genes, the attainment of normal SC number, and the support of meiotic and postmeiotic germ cell development. PMID:15761038

Tan, Karen A L; De Gendt, Karel; Atanassova, Nina; Walker, Marion; Sharpe, Richard M; Saunders, Philippa T K; Denolet, Evi; Verhoeven, Guido

2005-06-01

116

Co-culture of spermatogonial stem cells with sertoli cells in the presence of testosterone and FSH improved differentiation via up-regulation of post meiotic genes.  

PubMed

Spermatogonial stem cells (SSCs) maintain spermatogenesis throughout life in the male. Maintenance of SSCs and induction of spermiogenesis in vitro may provide a therapeutic strategy to treat male infertility. This study investigated in vitro differentiation of mouse SSCs in presence or absence of Sertoli cells, hormones and vitamins. Spermatogonial populations were enriched from testes of 4-6 week old males by magnetic activated cell sorting and anti-Thy-1 antibody. Sertoli cells isolated from 6-8 week old testes were enriched using lectin-DSA-coated plates. Isolated SSCs were cultured in the presence of Leukemia inhibitory factor (LIF) for 7 days in gelatin-coated dishes, then dissociated and cultured for 7 days in media lacking LIF in the presence or absence of Sertoli cells, with or without FSH, testosterone and vitamins. After one week, the effects of Sertoli cells ± supplementary media on SSC differentiation was evaluated by microscopy and expression of meiotic and postmeiotic transcripts using RT-PCR. SSC colonies had limited development after LIF removal alone, exhibiting low expression of meiotic (Scp3, Th2b) but not postmeiotic transcript, and loss of Stra8 and Dazl expression. SSCs co-cultured with Sertoli cells, hormones and vitamins developed spermatid-like cells expressing postmeiotic markers (TP1, TP2, Prm1) at levels over 2-fold higher than Sertoli cells or hormone/vitamins alone. Our present SSC-Sertoli co-culture provides conditions that may allow efficient in vitro differentiation of SSCs for the treatment of male infertility. PMID:23456578

Minaee Zanganeh, Bagher; Rastegar, Tayebeh; Habibi Roudkenar, Mehryar; Ragerdi Kashani, Iraj; Amidi, Fardin; Abolhasani, Farid; Barbarestani, Mohammad

2013-01-01

117

Malignant ovarian Sertoli-Leydig cell tumor localized with selective ovarian vein sampling.  

PubMed

Sertoli-Leydig cell tumors (SLCT) are rare, comprising less than 0.5% of ovarian neoplasms. They are most often diagnosed in premenopausal women and may produce androgens, resulting in hirsuitism, voice deepening, frontal balding, terminal hair growth, and clitoromegaly. SLCT are malignant in 15%-20% of cases. We discuss a 25-year-old patient with persistent hyperandrogenemia. Noninvasive imaging cannot conclusively differentiate between SCLT and other diagnoses such as polycystic ovary syndrome, ovarian hyperthecosis, idiopathic hyperandrogenism, idiopathic hirsuitism, and 21-hydroxylase-deficient nonclassic adrenal hyperplasia. Selective ovarian vein sampling revealed a 15-fold greater testosterone production from the right ovary compared with the left, which guided appropriate surgical management. PMID:23084689

Dunne, Caitlin; Havelock, Jon C

2012-01-01

118

Basigin null mutant male mice are sterile and exhibit impaired interactions between germ cells and Sertoli cells.  

PubMed

Basigin (BSG) is a multifunctional glycoprotein that plays an important role in male reproduction since male knockout (KO) mice are sterile. The Bsg KO testis lacks elongated spermatids and mature spermatozoa, a phenotype similar to that of alpha-mannosidase IIx (MX) KO mice. MX regulates formation of N-acetylglucosamine (GlcNAc) terminated N-glycans that participate in germ cell-Sertoli cell adhesion. Results showed that Bsg KO spermatocytes displayed normal homologous chromosome synapsis and progression through meiosis. However, only punctate expression of the round spermatid marker SP-10 in the acrosomal granule of germ cells of Bsg KO mice was detected indicating that spermatogenesis in Bsg KO mice was arrested at the early round spermatid stages. We observed a large increase in the number of germ cells undergoing apoptosis in Bsg KO testes. Using lectin blotting, we determined that GlcNAc terminated N-glycans are linked to BSG. GlcNAc terminated N-glycans were significantly reduced in Bsg KO testes. These observations indicate that BSG may act as a germ cell-Sertoli cell attachment molecule. Loss of BSG significantly reduced adhesion between GC-2 and SF7 cells. Moreover, wild type testes showed strong expression of N-cadherin (CDH2) while expression was greatly reduced in the testes of Bsg KO mice. In addition, the integrity of the blood-testis barrier (BTB) was compromised in Bsg KO testes. In conclusion, although some Bsg KO spermatogonia can undergo normal progression to the spermatocyte stage, BSG-mediated germ cell-Sertoli cell interactions appear to be necessary for integrity of the BTB and spermatocyte progression to mature spermatozoa. PMID:23727514

Bi, Jiajia; Li, Yanfen; Sun, Fengyun; Saalbach, Anja; Klein, Claudia; Miller, David J; Hess, Rex; Nowak, Romana A

2013-08-15

119

Therapy of experimental type 1 diabetes by isolated Sertoli cell xenografts alone  

PubMed Central

Type I diabetes mellitus is caused by autoimmune destruction of pancreatic ? cells, and effective treatment of the disease might require rescuing ? cell function in a context of reinstalled immune tolerance. Sertoli cells (SCs) are found in the testes, where their main task is to provide local immunological protection and nourishment to developing germ cells. SCs engraft, self-protect, and coprotect allogeneic and xenogeneic grafts from immune destruction in different experimental settings. SCs have also been successfully implanted into the central nervous system to create a regulatory environment to the surrounding tissue which is trophic and counter-inflammatory. We report that isolated neonatal porcine SC, administered alone in highly biocompatible microcapsules, led to diabetes prevention and reversion in the respective 88 and 81% of overtly diabetic (nonobese diabetic [NOD]) mice, with no need for additional ? cell or insulin therapy. The effect was associated with restoration of systemic immune tolerance and detection of functional pancreatic islets that consisted of glucose-responsive and insulin-secreting cells. Curative effects by SC were strictly dependent on efficient tryptophan metabolism in the xenografts, leading to TGF-?–dependent emergence of autoantigen-specific regulatory T cells and recovery of ? cell function in the diabetic recipients.

Fallarino, Francesca; Luca, Giovanni; Calvitti, Mario; Mancuso, Francesca; Nastruzzi, Claudio; Fioretti, Maria C.; Grohmann, Ursula; Becchetti, Ennio; Burgevin, Anne; Kratzer, Roland; van Endert, Peter; Boon, Louis; Calafiore, Riccardo

2009-01-01

120

Role of androgen and gonadotrophins in the development and function of the Sertoli cells and Leydig cells: Data from mutant and genetically modified mice  

Microsoft Academic Search

Development and maintenance of the male phenotype and establishment of fertility are all dependent upon the activity of the Sertoli cells and Leydig cells of the testis. This review examines the regulation and function of these cell during fetal and post-natal development. Fetal Leydig cells are sensitive to both luteinising hormone (LH) and adrenocorticotrophic hormone (ACTH) but Leydig cell function

P. J. O'Shaughnessy; I. D. Morris; I. Huhtaniemi; P. J. Baker; M. H. Abel

2009-01-01

121

Follicle-stimulating hormone induces hyperpolarization of immature rat Sertoli cells in monolayer culture.  

PubMed Central

1. The effect of FSH on the membrane potential of Sertoli cells has been investigated by intracellular microelectrode recording from monolayer cultures of cells, isolated from immature rat testes by enzymatic treatment. 2. In standard Earle's solution, the membrane potential of unstimulated cells in a 3-day-old monolayer was -21.6 +/- 0.2 mV (n = 300). The recorded potentials were unchanged on varying the culture period from 3 to 7 days or by removal of chloride or calcium from the media. However, they were decreased in a low-sodium or potassium-rich medium. 3. Ovine FSH caused hyperpolarization of the cell in a dose-dependent manner. Maximal values were obtained with concentrations ranging between 2.9 and 5.9 micrograms/ml (-37.0 +/- 0.2 mV; n = 310). Dibutyryl cyclic AMP (1 mM) produced a similar effect to FSH. Under similar conditions human chorionic gonadotrophin (hCG) had no effect on the membrane potential. 4. The FSH-induced hyperpolarization was unchanged by chloride or bicarbonate replacement. It was decreased by low-sodium media (9 mM) (-29.6 +/- 0.3 mV; n = 110), by removal of calcium (-32.4 +/- 0.6 mV; n = 128) and by an increase in the potassium concentration of the bathing medium. A tenfold increase in potassium concentration depolarized the cell by 29.5 mV against 16 mV for unstimulated cells. 5. FSH-induced hyperpolarization was decreased by application of ouabain (10(-4) M), quinidine (1.4 x 10(-4) M) and cobalt (10(-3) M) (respectively: -28.7 +/- 0.3 mV, n = 124; -26.6 +/- 0.3 mV, n = 52 and -24.8 +/- 0.3 mV, n = 68) but was unchanged by amiloride (10(-4) M) and TTX (3 x 10(-6) M). None of these drugs modified the membrane potential of unstimulated cells. 6. All these results suggest that FSH stimulation of Sertoli cells in monolayer culture involves the modification of their membrane permeabilities. Images Plate 1

Joffre, M; Roche, A

1988-01-01

122

Phorbol myristate acetate induces changes on F-actin and vinculin content in immature rat Sertoli cells  

Microsoft Academic Search

Actin and vinculin are two of the most abundant cytoskeletal proteins, widely expressed in nearly all types of eukaryotic cells. It has been well established that long-term exposure to the tumor promoter phorbol myristate acetate (PMA) affects Sertoli cell morphology, as well as F-actin and vinculin organization in vitro. To analyze in a quantitative manner the F-actin and vinculin content

M Kouloukoussa; V Aleporou-Marinou; B Angelopoulou; I. P Trougakos; E Panagopoulou; Chr Kittas; Evangelos Marinos

2004-01-01

123

Characterization of stage-specific tyrosinated ?-tubulin immunoperoxidase staining patterns in Sertoli cells of rat seminiferous tubules by light microscopic image analysis  

Microsoft Academic Search

Microtubules are involved in many structural and functional changes that occur in Sertoli cells during the cycle of the seminiferous epithelium. However, few studies have addressed stage-specific changes in the distribution of microtubules that accompany the process of spermatogenesis. This study provides a stage-by-stage immunohistochemical evaluation of Sertoli cell microtubules in paraffin sections of whole rat testes using an antibody

John R. Wenz; Rex A. Hess

1998-01-01

124

All-trans retinoic acid induces free radical generation and modulate antioxidant enzyme activities in rat sertoli cells.  

PubMed

In this work we investigated the effects of retinoic acid (RA) in Sertoli cells. Sertoli cells isolated from 15-day-old Wistar rats were previously cultured for 48 h and then treated with RA for 24 h. RA at high doses (1-10 microM) increased TBARS levels and induced a decrease in cell viability. At low doses (0.1-100 nM) RA did not increase TBARS level. RA also did not increase cell death at these doses. In order to investigate changes in antioxidant defenses we measured the CAT, SOD and GPx activities in Sertoli cells treated with RA. Compared to control, RA increased around 200% SOD activity in all doses tested (0.1-100 nM); GPx activity was increased 407.49, 208.98 and 243.88% (0.1, 1 and 10 nM, respectively); CAT activity was increased 127% with RA 1 nM. To clarify if RA induces ROS production per se, we performed experiments in vitro using 2-deoxyribose as specific substrate of oxidative degradation by *OH radical as well as TRAP assay. RA at 10 microM increased 2-deoxyribose degradation, suggesting that some of the RA-induced effects are mediated via *OH formation. Furthermore, the total reactive antioxidant potential (TRAP) of the RA was determined. At low concentrations RA has induced no redox activity. Conversely, higher concentration of RA (1-10 microM) increased chemiluminescence. The chemiluminescence produced was directly proportional to radical generation. We provide, for the first time, evidence for a free radical generation by RA. Our results demonstrated that RA plays an important role in Sertoli cells and these effects appear to be mediated by ROS. PMID:16479320

Conte da Frota, Mario Luiz; Gomes da Silva, Evandro; Behr, Guilherme Antônio; Roberto de Oliveira, Marcos; Dal-Pizzol, Felipe; Klamt, Fábio; Moreira, José Cláudio Fonseca

2006-04-01

125

A chimerical phagocytosis model reveals the recruitment by Sertoli cells of autophagy for the degradation of ingested illegitimate substrates.  

PubMed

Phagocytosis and autophagy are typically dedicated to degradation of substrates of extrinsic and intrinsic origins respectively. Although overlaps between phagocytosis and autophagy were reported, the use of autophagy for ingested substrate degradation by nonprofessional phagocytes has not been described. Blood-separated tissues use their tissue-specific nonprofessional phagocytes for homeostatic phagocytosis. In the testis, Sertoli cells phagocytose spermatid residual bodies produced during germ cell differentiation. In the retina, pigmented epithelium phagocytoses shed photoreceptor tips produced during photoreceptor renewal. Spermatid residual bodies and shed photoreceptor tips are phosphatidylserine-exposing substrates. Activation of the tyrosine kinase receptor MERTK, which is implicated in phagocytosis of phosphatidylserine-exposing substrates, is a common feature of Sertoli and retinal pigmented epithelial cell phagocytosis. The major aim of our study was to investigate to what extent phagocytosis by Sertoli cells may be tissue specific. We analyzed in Sertoli cell cultures that were exposed to either spermatid residual bodies (legitimate substrates) or retina photoreceptor outer segments (illegitimate substrates) the course of the main phagocytosis stages. We show that whereas substrate binding and ingestion stages occur similarly for legitimate or illegitimate substrates, the degradation of illegitimate but not of legitimate substrates triggers autophagy as evidenced by the formation of double-membrane wrapping, MAP1LC3A-II/LC3-II clustering, SQSTM1/p62 degradation, and by marked changes in ATG5, ATG9 and BECN1/Beclin 1 protein expression profiles. The recruitment by nonprofessional phagocytes of autophagy for the degradation of ingested cell-derived substrates is a novel feature that may be of major importance for fundamentals of both apoptotic substrate clearance and tissue homeostasis. PMID:23439251

Yefimova, Marina G; Messaddeq, Nadia; Harnois, Thomas; Meunier, Annie-Claire; Clarhaut, Jonathan; Noblanc, Anaïs; Weickert, Jean-Luc; Cantereau, Anne; Philippe, Michel; Bourmeyster, Nicolas; Benzakour, Omar

2013-05-01

126

Weight reduction and pioglitazone ameliorate polycystic ovary syndrome after removal of a Sertoli-stromal cell tumor  

PubMed Central

This report presents an unusual case of Sertoli-stromal cell tumor and polycystic ovary syndrome successfully treated with weight reduction and an insulin-sensitizing agent. A 22-year-old woman, gravida 0, para 0, visited our hospital for the first time with a 12-year history of secondary amenorrhea and hypertrichosis. Transvaginal ultrasonography revealed a solid tumor in the right ovary. Right salpingo-oophorectomy was performed and pathological examination confirmed a Sertoli-stromal cell tumor. The patient’s serum androgen levels declined postoperatively, but remained above normal. Pioglitazone treatment for 6 months also significantly reduced serum androgen levels, but they still remained above normal. However, after losing 12 kg of body weight, the patient’s serum androgen levels declined to normal, and spontaneous menstruation became regular. Weight reduction with pioglitazone is an effective means of treating hyperandrogenism.

Baba, Tsuyoshi; Endo, Toshiaki; Ikeda, Keiko; Shimizu, Ayumi; Morishita, Miyuki; Kuno, Yoshika; Honnma, Hiroyuki; Kiya, Tamotsu; Ishioka, Shin-ichi; Saito, Tsuyoshi

2012-01-01

127

Testicular dysgenesis does not affect expression of anti-m?llerian hormone by Sertoli cells in premeiotic seminiferous tubules.  

PubMed Central

Anti-Müllerian hormone (AMH) immunoreactivity was studied on paraffin sections obtained from archival testicular biopsies of 29 children with intersex disorders and of 22 controls. Strong AMH immunoreactivity was observed in Sertoli cell cytoplasm from 8 fetal weeks until puberty. During pubertal maturation, in both normal and intersex patients, AMH expression was present in premeiotic seminiferous tubules, but was no longer detected in neighboring tubules with meiotic development. AMH immunostaining was abolished in the testis of one patient with persistent Müllerian ducts due to a mutation of the AMH gene, but was conserved in the testes of two patients with mutations of the AMH receptor gene. Testicular dysgenesis usually results in sexual ambiguity, with low testosterone and AMH serum levels and persistence of Müllerian derivatives. AMH immunoreactivity was conserved in premeiotic seminiferous tubules of dysgenetic testes, and also in sex-cord cells of a gonadoblastoma. In patients with asymmetric gonadal differentiation, the streak gonad was AMH-negative. In conclusion, secretion of AMH is a constitutive feature of the immature Sertoli cell and its expression is altered only by mutations of the AMH gene, but not by gonadal dysgenesis. The degree of regression of Müllerian ducts and serum AMH levels reflect the number, not the functional value, of Sertoli cells present in the immature testis. Images Figure 1 Figure 2 Figure 3

Rey, R.; al-Attar, L.; Louis, F.; Jaubert, F.; Barbet, P.; Nihoul-Fekete, C.; Chaussain, J. L.; Josso, N.

1996-01-01

128

Trafficking of sulfated glycoprotein-1 (prosaposin) to lysosomes or to the extracellular space in rat Sertoli cells  

Microsoft Academic Search

Sulfated glycoprotein-1 (prosaposin) exists in 2 forms: a 65rkDa form targeted to lysosomes and a 70rkDa form secreted extracellularly. In order to understand the sorting and targeting mechanisms of the two forms of SGP-1, we have compared their maturation, processing, and secretion in rat Sertoli cells in vivo. Metabolic labeling experiments in vivo demonstrated that the 65rkDa form is synthesized

Suleiman A. Igdoura; Adam Rasky; Carlos R. Morales

1996-01-01

129

The ATP-binding cassette transporter 1 mediates lipid efflux from Sertoli cells and influences male fertility  

Microsoft Academic Search

The liver X receptor\\/retinoid X receptor (LXR\\/ RXR)-regulated gene ABCA1 effluxes cellular cholesterol and phospholipid to apolipoprotein A1 (apoA1), which is the rate-limiting step in high-density lipoprotein synthesis. The RXR pathway plays a critical role in testicular lipid traf- ficking, and RXR ? -deficient male mice are sterile and accu- mulate lipids in Sertoli cells. Here, we demonstrate that ABCA1

David M. Selva; Veronica Hirsch-Reinshagen; Braydon Burgess; Steven Zhou; Jeniffer Chan; Sean McIsaac; Michael R. Hayden; Geoffrey L. Hammond; A. Wayne Vogl; Cheryl L. Wellington

2004-01-01

130

Effects of Relatively Low Levels of Mono(2-Ethylhexyl) Phthalate on Cocultured Sertoli Cells and Gonocytes from Neonatal Rats  

Microsoft Academic Search

Di-(2-ethylhexyl) phthalate (DEHP), one of the abundant man-made environmental chemicals, induces testicular damage in both developing and adult animals. However, the nature and mechanism underlying the action of phthalates on testicular development remain largely unexplored. In the present study, we used cocultures of neonatal Sertoli cells and gonocytes (precursors of spermatogonia) to characterize in detail the effects of mono-(2-ethylhexyl) phthalate

Ling-Hong Li; William F. Jester; Joanne M. Orth

1998-01-01

131

Altered testicular development as a consequence of increase number of sertoli cell in male lambs exposed prenatally to excess testosterone.  

PubMed

The reprograming effects of prenatal testosterone (T) treatment on postnatal reproductive parameters have been studied extensively in females of several species but similar studies in males are limited. We recently found that prenatal T treatment increases Sertoli cell number and reduced spermatogenesis in adult rams. If such disruptions are manifested early in life and involve changes in testicular paracrine environment remain to be explored. This study addresses the impact of prenatal T excess on testicular parameters in infant males, including Sertoli cell number and expression of critical genes [FSH receptor (FSHR), androgen receptor (AR), transforming growth factor beta 1 (TGFB1), 3 (TGFB3), transforming growth factor beta type 1 receptor, (TGFBR1), and anti-Müllerian hormone (AMH)] modulating testicular function. At 4 week of age, male lambs born to dams treated with 30 mg of T propionate twice weekly from day 30 to 90, followed by 40 mg of T propionate from day 90 to 120 of pregnancy (T-males), had a higher number of Sertoli cells/testis (P = 0.035) than control males (C-males) born to dams treated with the vehicle. While no differences were observed in the expression of FSHR and TGFB3, testicular TGFBR1 expression was found to be lower in T-males (P = 0.03) compared to C-males. Expression level of AMH, TGFB1, and AR also tended to be lower in T-males. These findings provide evidence that impact of fetal exposure to T excess is evident early in postnatal life, mainly characterized by an increase in Sertoli cell number. This could explain the testicular dysfunction observed in adult rams. PMID:23076741

Rojas-García, Pedro P; Recabarren, Mónica P; Sir-Petermann, Teresa; Rey, Rodolfo; Palma, Sergio; Carrasco, Albert; Perez-Marin, Carlos C; Padmanabhan, Vasantha; Recabarren, Sergio E

2013-06-01

132

Retinoblastoma protein (RB) interacts with E2F3 to control terminal differentiation of Sertoli cells.  

PubMed

The retinoblastoma protein (RB) is essential for normal cell cycle control. RB function depends, at least in part, on interactions with the E2F family of DNA-binding transcription factors (E2Fs). To study the role of RB in the adult testis, a Sertoli cell (SC)-specific Rb knockout mouse line (SC-RbKO) was generated using the Cre/loxP recombination system. SC-RbKO mice exhibited an age-dependent testicular atrophy, impaired fertility, severe SC dysfunction, and spermatogenic defects. Removal of Rb in SC induced aberrant SC cycling, dedifferentiation, and apoptosis. Here we show that E2F3 is the only E2F expressed in mouse SCs and that RB interacts with E2F3 during mouse testicular development. In the absence of RB, the other retinoblastoma family members p107 and p130 began interacting with E2F3 in the adult testes. In vivo silencing of E2F3 partially restored the SC maturation and survival as well as spermatogenesis in the SC-RbKO mice. These results point to RB as a key regulator of SC function in adult mice and that the RB/E2F3 pathway directs SC maturation, cell cycle quiescence, and RB protects SC from apoptosis. PMID:24901045

Rotgers, E; Rivero-Müller, A; Nurmio, M; Parvinen, M; Guillou, F; Huhtaniemi, I; Kotaja, N; Bourguiba-Hachemi, S; Toppari, J

2014-01-01

133

Frequent mutation and nuclear localization of ?-catenin in sertoli cell tumors of the testis.  

PubMed

The Sertoli cell tumor (SCT) of the testis is a sex cord stromal tumor, usually sporadic, rarely associated with genetic syndromes. Much remains unclear about the molecular genetic changes involved in SCT and its histogenesis. Recently, nuclear ?-catenin immunostaining has been reported in a case of bilateral SCT, but the molecular basis of the aberrant nuclear ?-catenin expression remains uncertain. In the present study, ?-catenin immunohistochemical assay and mutational analysis of exon 3 of the CTNNB1 gene by direct sequencing were performed in 14 SCTs, 2 of which had an unfavorable clinical course. Immunohistochemical study showed that ?-catenin was located in the cytoplasm of tumor cells in 4 cases (28.6%) and in both the nuclei and the cytoplasm in the remaining 10 cases (71.4%). ?-Catenin mutations were detected in 10 of the 14 patients (71.4%) under evaluation. Ten of 10 mutation-carrying cases showed strong nuclear and diffuse cytoplasmic ?-catenin immunoreactivity. Seven of the 8 CTNNB1-mutated tumors tested for cyclin D1 displayed diffuse immunoreactivity in the nuclei of tumor cells. We conclude that CTNNB1 exon 3 mutations are likely to be involved in the pathogenesis of male SCT with nuclear accumulation of ?-catenin and affect the expression of cyclin D1. PMID:24061522

Perrone, Federica; Bertolotti, Alessia; Montemurro, Gabriella; Paolini, Biagio; Pierotti, Marco A; Colecchia, Maurizio

2014-01-01

134

Copy Number Variants in Patients with Severe Oligozoospermia and Sertoli-Cell-Only Syndrome  

PubMed Central

A genetic origin is estimated in 30% of infertile men with the common phenotypes of oligo- or azoospermia, but the pathogenesis of spermatogenic failure remains frequently obscure. To determine the involvement of Copy Number Variants (CNVs) in the origin of male infertility, patients with idiopathic severe oligozoospermia (N?=?89), Sertoli-cell-only syndrome (SCOS, N?=?37)) and controls with normozoospermia (N?=?100) were analysed by array-CGH using the 244A/400K array sets (Agilent Technologies). The mean number of CNVs and the amount of DNA gain/loss were comparable between all groups. Ten recurring CNVs were only found in patients with severe oligozoospermia, three only in SCOS and one CNV in both groups with spermatogenic failure but not in normozoospermic men. Sex-chromosomal, mostly private CNVs were significantly overrepresented in patients with SCOS. CNVs found several times in all groups were analysed in a case-control design and four additional candidate genes and two regions without known genes were associated with SCOS (P<1×10?3). In conclusion, by applying array-CGH to study male infertility for the first time, we provide a number of candidate genes possibly causing or being risk factors for the men's spermatogenic failure. The recurring, patient-specific and private, sex-chromosomal CNVs as well as those associated with SCOS are candidates for further, larger case-control and re-sequencing studies.

Tuttelmann, Frank; Simoni, Manuela; Kliesch, Sabine; Ledig, Susanne; Dworniczak, Bernd; Wieacker, Peter; Ropke, Albrecht

2011-01-01

135

Combined Leydig cell and Sertoli cell dysfunction in 46,XX males lacking the sex determining region Y gene.  

PubMed

We have evaluated 3 individuals with a rare form of 46,XX sex reversal. All of them had ambiguous external genitalia and mixed wolffian and müllerian structures, indicating both Leydig cell and Sertoli cell dysfunction, similar to that of patients with true hermaphroditism. However, gonadal tissue was not ovotesticular but testicular with varying degrees of dysgenesis. SRY sequences were absent in genomic DNA from peripheral leukocytes in all 3 subjects. Y centromere sequences were also absent, indicating that testis development did not occur because of a low level mosaicism of Y bearing cells. The subjects in this report demonstrate that there is a continuum in the extent of testis determination in SRY-negative 46,XX sex reversal, ranging from nearly normal to minimal testicular development. PMID:7677147

Turner, B; Fechner, P Y; Fuqua, J S; Marcantonio, S M; Perlman, E J; Vordermark, J S; Berkovitz, G D

1995-07-01

136

Sertoli cells control peritubular myoid cell fate and support adult Leydig cell development in the prepubertal testis.  

PubMed

Sertoli cells (SCs) regulate testicular fate in the differentiating gonad and are the main regulators of spermatogenesis in the adult testis; however, their role during the intervening period of testis development, in particular during adult Leydig cell (ALC) differentiation and function, remains largely unknown. To examine SC function during fetal and prepubertal development we generated two transgenic mouse models that permit controlled, cell-specific ablation of SCs in pre- and postnatal life. Results show that SCs are required: (1) to maintain the differentiated phenotype of peritubular myoid cells (PTMCs) in prepubertal life; (2) to maintain the ALC progenitor population in the postnatal testis; and (3) for development of normal ALC numbers. Furthermore, our data show that fetal LCs function independently from SC, germ cell or PTMC support in the prepubertal testis. Together, these findings reveal that SCs remain essential regulators of testis development long after the period of sex determination. These findings have significant implications for our understanding of male reproductive disorders and wider androgen-related conditions affecting male health. PMID:24803659

Rebourcet, Diane; O'Shaughnessy, Peter J; Pitetti, Jean-Luc; Monteiro, Ana; O'Hara, Laura; Milne, Laura; Tsai, Yi Ting; Cruickshanks, Lyndsey; Riethmacher, Dieter; Guillou, Florian; Mitchell, Rod T; van 't Hof, Rob; Freeman, Tom C; Nef, Serge; Smith, Lee B

2014-05-01

137

NC1 domain of collagen ?3(IV) derived from the basement membrane regulates Sertoli cell blood-testis barrier dynamics  

PubMed Central

The blood-testis barrier (BTB) is an important ultrastructure for spermatogenesis. Delay in BTB formation in neonatal rats or its irreversible damage in adult rats leads to meiotic arrest and failure of spermatogonial differentiation beyond type A. While hormones, such as testosterone and FSH, are crucial to BTB function, little is known if there is a local regulatory mechanism in the seminiferous epithelium that modulates BTB function. Herein, we report that collagen ?3(IV) chain, a component of the basement membrane in the rat testis, could generate a noncollagenous (NC1) domain peptide [Col?3(IV) NC1] via limited proteolysis by matrix metalloproteinase-9 (MMP-9), and that the expression of MMP-9 was upregulated by TNF?. While recombinant Col?3(IV) NC1 protein produced in E. coli failed to perturb Sertoli cell tight junction (TJ)-permeability barrier function, possibly due to the lack of glycosylation, Col?3(IV) NC1 recombinant protein produced in mammalian cells and purified to apparent homogeneity by affinity chromatography was found to reversibly perturb the Sertoli cell TJ-barrier function. Interestingly, Col?3(IV) NC1 recombinant protein did not perturb the steady-state levels of several TJ- (e.g., occludin, CAR, JAM-A, ZO-1) and basal ectoplasmic specialization- (e.g., N-cadherin, ?-catenin, ?-catenin) proteins at the BTB but induced changes in protein localization and/or distribution at the Sertoli cell-cell interface in which these proteins moved from the cell surface into the cell cytosol, thereby destabilizing the TJ function. These findings illustrate the presence of a local regulatory axis known as the BTB-basement membrane axis that regulates BTB restructuring during spermatogenesis.

Wong, Elissa W.P.; Cheng, C. Yan

2013-01-01

138

Activation of innate immune system in response to lipopolysaccharide in chicken Sertoli cells.  

PubMed

Sertoli cells (SCs) play an important physiological role in the testis, as they support, nourish, and protect the germ cells. As protection of the developing spermatozoa is an emerging aspect of reproductive physiology, this study examined the expression pattern of innate immune-related genes, including avian ?-defensins (AvBDs), Toll-like receptors (TLRs), and cytokines, and investigated the time course of an inflammatory response in rooster SCs triggered by exposure to the bacterial endotoxin lipopolysaccharide (LPS). SCs were isolated from 6-week-old chicken, cultured in vitro, and stimulated with 1??g/ml LPS at different time courses (0, 6, 12, 24, and 48?h). Data on expression analysis revealed that all ten members of the chicken TLR family, nine members of the AvBD family, as well as eight cytokine genes were expressed in SCs. Quantitative real-time PCR analysis revealed that LPS treatment resulted in significant induction of the expression levels of six TLRs, six AvBDs, and four cytokine genes, while two cytokine genes were downregulated and two other genes were unchanged. The increasing interleukin 1? (IL1?) production was confirmed in the conditioned medium. Furthermore, the phagocytosis of SCs was increased after LPS treatment. In conclusion, these findings provide evidence that SCs express innate immune-related genes and respond directly to bacterial ligands. These genes represent an important component of the immune system, which could be integrated into semen, and present a distinctive constituent of the protective repertoire of the testis against ascending infections. PMID:24920664

Michailidis, Georgios; Anastasiadou, Maria; Guibert, Edith; Froment, Pascal

2014-09-01

139

Expression characterization of testicular DMRT1 in both Sertoli cells and spermatogenic cells of polyploid gibel carp.  

PubMed

Dmrt1 has been suggested to play significant roles in sex determination and differentiation, but various expression patterns and cell types have been observed in the testis of vertebrates. Polyploid gibel carp, because of the multiple modes of unisexual gynogenesis and sexual reproduction, has become a unique case to explore the evolution of sex determination and differentiation. However, the sex-determination related genes in gibel carp have remained unknown. In this study, we identified and characterized 4 cDNAs of Dmrt1 genes. Subsequently, a polyclonal antibody specific to CagDMRT1 was prepared to examine its expression and distribution patterns at protein level. Significantly, both relative real-time PCR and Western blot detection confirmed predominant expression of CagDmrt1 in the adult testis of gibel carp. Moreover, the intensive expression of CagDMRT1 around spermatogenic cysts was revealed during spermatogenesis. And, following immunofluorescence co-localization of CagDMRT1 and CagVASA, a prominent CagDMRT1 expression in Sertoli cells and a mild CagDMRT1 expression in spermatogenic cells including spermatogonia and primary spermatocytes were clearly characterized. The CagDMRT1 signal in Sertoli cells is extensively distributed in both nuclei and cytoplasm, while the CagDMRT1 in spermatogonia and primary spermatocytes is mainly expressed in nuclei, and there is only the remained CagDMRT1 signal in the cytoplasm of secondary spermatocytes. These findings suggest that DMRT1 should be related to testis differentiation and spermatogenesis in gibel carp. PMID:25020260

Li, Xi-Yin; Li, Zhi; Zhang, Xiao-Juan; Zhou, Li; Gui, Jian-Fang

2014-09-10

140

Environmental Tobacco Smoke Exposure during Intrauterine Period, Promotes Caspase Dependent and Independent DNA Fragmentation in Sertoli-Germ Cells  

PubMed Central

Objectives. To investigate the effect of cigarette smoke exposure during intrauterine period on neonatal rat testis. Methods. Twenty-five rats were randomized to be exposed to cigarette smoke with the Walton Smoking Machine or to room air during their pregnancies. The newborn male rats (n = 21) were grouped as group 1 (n = 15) which were exposed to cigarette smoke during intrauterine life and group 2 (n = 6) which were exposed to room air during intrauterine life. The orchiectomy materials were analyzed with TUNEL immunofluorescent staining for detection of DNA damage. To detect apoptosis, immunohistochemical analyses with caspase-3 were performed. Primary outcomes were apoptotic index and immunohistochemical scores (HSCORES); secondary outcomes were Sertoli-cell count and birth-weight of rats. Results. Sertoli cell apoptosis was increased in group 1 (HSCORE = 210.6 ± 41.9) when compared to group 2 (HSCORE = 100.0 ± 17.8) (P = 0.001). Sertoli cell count was decreased in group 1 (P = 0.043). The HSCORE for the germ cells was calculated as 214.0 ± 46.2 in group 1 and 93.3 ± 10.3 in group 2 (P = 0.001) referring to an increased germ cell apoptosis in group 1. The apoptotic indexes for group 1 were 49.6 ± 9.57 and 29.98 ± 2.34 for group 2 (P = 0.001). The immunofluorescent technique demonstrated increased DNA damage in seminiferous epithelium in group 1. Conclusions. Intrauterine exposure to cigarette smoke adversely affects neonatal testicular structuring and diminishes testicular reserve.

Yuksel, Beril; Kilic, Sevtap; Lortlar, Nese; Tasdemir, Nicel; Sertyel, Semra; Bardakci, Yesim; Aksu, Tarik; Batioglu, Sertac

2014-01-01

141

Transfection with steroid-responsive reporter constructs shows glucocorticoid rather than androgen responsiveness in cultured Sertoli cells.  

PubMed

It remains unclear why it has proven so difficult to identify androgen target genes in cultured Sertoli cells. Given the lack of useful endogenous reporter genes, we studied the androgen and glucocorticoid responsiveness of these cells by transfection with three different steroid-responsive reporter constructs. The constructs were driven by the tyrosine aminotransferase steroid-responsive region (TAT-GRE4x-Luc), the mouse mammary tumor virus promoter (MMTV-Luc) and the Pem homeobox gene proximal promoter respectively (Pem-Luc). These constructs can be activated either by both the glucocorticoid receptor (GR) and the androgen receptor (AR) (TAT-GRE4x-Luc and MMTV-Luc) or selectively by the AR (Pem-Luc). Despite high transfection efficiency (30-40%) none of the constructs could be activated by treatment of the Sertoli cells with testosterone, 5alpha-dihydrotestosterone or synthetic androgens. Even pretreatment with follicle-stimulating hormone to raise AR levels (from 31 up to 82fmol/mg protein) did not result in androgen responsiveness. In contrast, treatment with dexamethasone markedly stimulated TAT-GRE4x-Luc and MMTV-Luc activity. GR levels reached a value of 172fmol/mg protein in the cultured cells and both AR and GR displayed homogeneous distribution by immunocytochemical evaluation. Androgen responsiveness was restored and glucocorticoid responsiveness was increased by cotransfection with AR or GR expression constructs. Under cotransfection conditions, 1nM of testosterone (a concentration that is some 100 times lower than that estimated to be present in the testis) was sufficient to stimulate the TAT-GRE4x-Luc maximally. Our data indicate that cultured Sertoli cells respond better to glucocorticoids than to androgens and that one of the factors limiting androgen responsiveness is the availability of AR. Other factors limiting the transactivation capacity of the (endogenous) AR, however, cannot be excluded. PMID:16388947

Denolet, Evi; Gendt, Karel De; Swinnen, Johannes V; Verrijdt, Guy; Deboel, Ludo; Roskams, Tania; Verhoeven, Guido

2006-02-01

142

Regulation of the phosphoinositide pathway in cultured Sertoli cells from immature rats: effects of follicle-stimulating hormone and fluoride  

SciTech Connect

Many hormones elicit effects on target cells by stimulating the enzyme phospholipase-C, which catalyzes the hydrolysis of phosphoinositides to the intracellular second messengers diacylglycerol and inositol phosphates. The present study examined the roles of FSH and guanine nucleotide-binding proteins (G-proteins) in regulating the hydrolysis of phosphoinositides in Sertoli cells. Sertoli cell cultures prepared from 16- to 18-day-old rats were incubated for 24 h with myo-(2-3H) inositol to label endogenous phospholipids. Treatment of cells from 0.5-20 min with preparations of ovine FSH ranging in potency from 1-60 times that of NIH FSH S1 did not affect accumulation of inositol phosphates. Levels of total (3H)inositol phosphates ((3H)inositol mono-, di-, and triphosphates (IP, IP2, and IP3)) in FSH-treated cultures was 75-120% the levels in control cultures over the various time intervals studied. Addition of testosterone and the combination of testosterone plus retinoic acid, agents that have been shown to potentiate effects of FSH in other systems, did not affect accumulation of inositol phosphates in response to FSH. In contrast to the lack of effect on accumulation of inositol phosphates, FSH stimulated 4- to 11-fold increases in estradiol secretion over 24 h of culture, indicating that Sertoli cells were viable and responsive to FSH. AIF4- has been shown to activate G-proteins involved in regulation of adenylate cyclase activity. In the present study, AIF4- induced 4- to 5-fold increases in IP, IP2, and IP3 in experiments wherein FSH had no effect. Pretreatment of Sertoli cells with pertussis toxin (100 and 1000 ng/ml) for 24 h inhibited fluoride-induced generation of IP, IP2, and IP3 by 24-51%. Similar treatment with cholera toxin had no effect on basal or fluoride-induced generation of IP2 or IP3, but increased fluoride-induced generation of IP by 20-34%.

Quirk, S.M.; Reichert, L.E. Jr.

1988-07-01

143

Organic Anion Transporter 6 (Slc22a20) Specificity and Sertoli Cell-Specific Expression Provide New Insight on Potential Endogenous Roles  

PubMed Central

Organic anion transporter 6 (Oat6; Slc22a20), a member of the OAT family, was demonstrated previously to mediate the transport of organic anions (Am J Physiol Renal Physiol 291:F314–F321, 2006). In the present study, we sought to further delineate the function of murine Oat6 (mOat6) by analyzing the effect of select organic anions on mOat6-mediated transport by using a Chinese hamster ovary (CHO) cell line stably expressing mOat6 (CHO-mOat6). When examined, kinetic analysis demonstrated that the mechanism of inhibition of mOat6 and mOat3 was competitive. Homovanillic acid, 5-hydroxyindole acetic acid, 2,4-dihydroxyphenylacetate, hippurate, and dehydroepiandrosterone sulfate (DHEAS) each significantly reduced mOat6 activity with inhibitory constant (Ki) values of 3.0 ± 0.5, 48.9 ± 10.3, 61.4 ± 7.1, 59.9 ± 4.9, and 38.8 ± 3.1 ?M, respectively. Comparison to Ki values determined for mOat3 (67.8 ± 7.2, 134.5 ± 27.0, 346.7 ± 97.9, 79.3 ± 4.0, and 3.8 ± 1.1 ?M, respectively) revealed that there are significant differences in compound affinity between each transporter. Fluoroquinolone antimicrobials and reduced folates were without effect on mOat6-mediated uptake. Investigation of testicular cell type-specific expression of mOat6 by laser capture microdissection and quantitative polymerase chain reaction revealed significant mRNA expression in Sertoli cells, but not in Leydig cells or spermatids. Overall, these data should aid further refinements to the interpretation and modeling of the in vivo disposition of OAT substrates. Specifically, expression in Sertoli cells suggests Oat6 may be an important determinant of blood-testis barrier function, with Oat6-mediated transport of estrone sulfate and DHEAS possibly representing a critical step in the maintenance of testicular steroidogenesis.

Schnabolk, Gloriane W.; Gupta, Bhawna; Mulgaonkar, Aditi; Kulkarni, Mrugaya

2010-01-01

144

Zinc and low-dose of cadmium protect sertoli cells against toxic-dose of cadmium: The role of metallothionein.  

PubMed

Background: The impact of cadmium (Cd) on male infertility may be related to the interaction with metal-binding proteins known as metallothioneins (Mts). Trace elements like zinc (Zn) have protective effects on testicular damage induced by Cd. Objective: We determined the effect of Zn and low-dose Cd pre-treatment on the expression of Mt1 and Mt2 genes on testicular Sertoli cells. Materials and Methods: The cultured TM4 mouse sertoli cells were treated with 50 ?M ZnSO4 (Zn pre-treated group; ZnPG), 2 ?M CdCl2 (Cd pre-treated group; CdPG), or distilled water (DW pre-treated group; DWPG). After 18 hour, all of these groups were exposed to 100 ?M CdCl2 for different periods of time (1, 2, 3, and 6 hours). There was also a control group for all three groups, which was treated only with distilled water (without Cd or Zn pre-treatment). Cellular viability, Zn and Cd concentrations and gene expression were assessed by MTT, atomic absorption spectrometry and real time PCR methods, respectively. Results: The expression of Mt1 and Mt2 genes in ZnPG, CdPG, and DWPG was greater than the control group (p=0.02 and p=0.01, respectively). Cd concentrations in CdPG and DWPG were greater than the control group (p=0.00). Expression of both genes in ZnPG and CdPG increased after 3 hours of treatment and Cd concentration decreased simultaneously, which was more obvious in ZnPG. Conclusion: Zn and short term low-dose Cd pre-treatment might reduce the adverse effects of Cd by increasing expression of Mts genes in Sertoli cells. The protective effect of Zn was stronger than Cd. PMID:24639783

Kheradmand, Fatemeh; Nourmohammadi, Issa; Ahmadi-Faghih, Mohamad Amin; Firoozrai, Mohsen; Modarressi, Mohammad Hossein

2013-06-01

145

Regulatory influence of germ cells on sertoli cell function in the pre-pubertal rat after acute irradiation of the testis.  

PubMed

While germ cell regulation of Sertoli cells has been extensively explored in adult rats in vivo, in contrast, very little is known about germ cell influence on Sertoli cell function at the time when spermatogenesis begins and develops. In the present study various Sertoli cell parameters (number, testicular androgen binding protein (ABP) and testin, serum inhibin-B and, indirectly, follicle-stimulating hormone (FSH)) were investigated after the exposure of 19-day-old rats to a low dose of 3 Grays of gamma-rays. Differentiated spermatogonia were the primary testicular targets of the gamma-rays, which resulted in progressive maturation depletion, sequentially and reversibly affecting all germ cell classes. Testicular weight declined to a nadir when pachytene spermatocytes and spermatids were depleted from the seminiferous epithelium and complete or near complete recovery of spermatogenesis and testicular weight was observed at the end of the experiment. Blood levels of FSH and ABP were normal during the first 11 days after irradiation, when spermatogonia and early spermatocytes were depleted. While the number of Sertoli cells was not significantly affected by the irradiation, from days 11-66 after gamma-irradiation, ABP production declined and FSH levels increased when pachytene spermatocytes and spermatids were depleted and the recovery of these parameters was only observed when spermatogenesis was fully restored. Comparison of the pattern of change in serum levels of inhibin-B and testicular levels of testin and of germ cell numbers strongly suggest a relationship between the disappearance of spermatocytes and spermatids from the seminiferous epithelium and the decrease in levels of inhibin-B and increase in levels of testin from 7 to 36 days post-irradiation. Levels of testin and inhibin-B were restored before spermatogenesis had totally returned to normal. In conclusion, this in vivo study shows that pre-pubertal Sertoli cell function is under the complex control of various germ cell classes. This control presents clear differences when compared with that previously observed in adult animals and depends on the Sertoli cell parameter of interest, as well as on the germ cell type. PMID:11114978

Guitton, N; Touzalin, A M; Sharpe, R M; Cheng, C Y; Pinon-Lataillade, G; Méritte, H; Chenal, C; Jégou, B

2000-12-01

146

Xanthine oxidase-dependent ROS production mediates vitamin A pro-oxidant effects in cultured Sertoli cells.  

PubMed

Several studies have suggested that vitamin A (retinol, ROH) presents pro-oxidant properties in biological systems. Recent studies point out that xantine oxidase, a ROS-generating enzyme, catalyses ROH oxidation to RA in vitro. These works stimulated the authors to investigate whether xanthine oxidase could be involved on the ROH pro-oxidative effects reported in cultured Sertoli cells. In vitro, it was demonstrate that xanthine oxidase generates superoxide in the presence of ROH as assessed by superoxide mediated-NBT reduction. Superoxide production is potentiated in the presence of NADH and inhibited by allopurinol. In Sertoli cells, ROH treatment increased xanthine oxidase activity and inhibition of the enzyme with allopurinol attenuated ROH-induced ROS production, protein damage and cytotoxicity. Moreover, inhibition of ROH oxidation to RA by retinaldehyde dehydrogenase inhibitor potentiated both xanthine oxidase-dependent ROS production and cell damage in ROH-treated cells. The data show that xanthine oxidase may play a role on vitamin A pro-oxidant effects. PMID:18569017

Zanotto-Filho, Alfeu; Schröder, Rafael; Moreira, José Cláudio F

2008-06-01

147

Identification of genetic networks involved in the cell injury accompanying endoplasmic reticulum stress induced by bisphenol A in testicular Sertoli cells  

Microsoft Academic Search

To identify detailed mechanisms by which bisphenol A (BPA), an endocrine-disrupting chemical, induces cell injury in mouse testicular Sertoli TTE3 cells, we performed genome-wide microarray and computational gene network analyses. BPA (200?M) significantly decreased cell viability and simultaneously induced an increase in mRNA levels of HSPA5 and DDIT3, endoplasmic reticulum (ER) stress marker genes. Of the 22,690 probe sets analyzed,

Yoshiaki. Tabuchi; Ichiro Takasaki; Takashi Kondo

2006-01-01

148

Bilateral Laparoscopic Gonadectomy in a Patient With Complete Androgen Insensitivity Syndrome and Bilateral Sertoli-Leydig Cell Tumor: A Case Report and Brief Review of the Literature  

PubMed Central

Introduction: Complete androgen insensitivity syndrome (previously called testicular feminization) is specified by a 46 XY karyotype and negative sex chromatin, bilateral undescended testes, female genitalia appearance, and lack of mullerian derivatives. Case Presentation: A 28-year-old woman with complete (severe) androgen resistance underwent prophylactic laparoscopic bilateral gonadectomy because of the eventually increased risk of gonadal malignancy. Although the gonads appeared grossly normal, microscopic examination revealed bilateral well differentiated sertoli–leydig cell tumor (SLCT). Discussion: Our Medline search revealed that this is the first reported case of bilateral sertoli–leydig cell tumor (SLCT) in androgen insensitivity syndrome.

Asl Zare, Mohammad; Kalantari, Mahmood Reza; Asadpour, Amir Abbas; Kamalati, Ali

2014-01-01

149

Poorly Differentiated Ovarian Sertoli-Leydig Cell Tumor in a 16-Year-Old Single Woman: A Case Report and Literature Review  

PubMed Central

Sertoli-Leydig cell tumor (SLCT) of ovary is an exceedingly unusual neoplasm that belongs to a group of sex cord-stromal tumors of ovary and accounts for less than 0.5% of all primary ovarian neoplasms. Very few case reports have been documented in the literature so far. Herein, we report a case of primary poorly differentiated ovarian Sertoli-Leydig cell tumor (SLCT) involving the left ovary in a 16-year-old single woman who presented with a 3-month history of a pelviabdominal mass, acne, hirsutism, and menstrual irregularities. In addition, a literature review on ovarian SLCTs is provided.

Abu-Zaid, Ahmed; Azzam, Ayman; Alghuneim, Lama Abdulhamid; Metawee, Mona Tarek; Amin, Tarek; Al-Hussain, Turki Omar

2013-01-01

150

The Sertoli Cell Only Syndrome and Glaucoma in a Sex - Determining Region Y (SRY) Positive XX Infertile Male  

PubMed Central

The XX male syndrome is a rare genetic disorder. The phenotype is variable; it ranges from a severe impairment of the external genitalia to a normal male phenotype with infertility. It generally results from an unequal crossing over between the short arms of the sex chromosomes (X and Y). We are reporting a case of a 38-year-old man who presented with infertility and the features of hypogonadism and glaucoma. The examinations revealed normal external male genitalia, soft small testes, gynaecomastia and glaucoma. The semen analysis showed azoospermia. The serum gonadotropins were high, with low Anti Mullerian Hormone (AMH) and Inhibin B levels. The chromosomal analysis demonstrated a 46, XX karyotype. Fluorescent In-Situ Hybridization (FISH) and Polymerase Chain Reaction (PCR) revealed the presence of a Sex-determining Region Y (SRY). Testicular Fine Needle Aspiration Cytology (FNAC) revealed the Sertoli Cell Only Syndrome (SCOS). The presence of only Sertoli Cells in the testes, with glaucoma in the XX male syndrome, to our knowledge, has not been reported in the literature.

Jain, Manish; V, Veeramohan; Chaudhary, Isha; Halder, Ashutosh

2013-01-01

151

Crlz-1 is prominently expressed in spermatogonia and Sertoli cells during early testis development and in spermatids during late spermatogenesis.  

PubMed

The expression of the Crlz-1 gene in mouse testis, where it was found to be expressed most highly among the tested mouse organs, was analyzed spatiotemporally by employing RT-PCR and in situ hybridization techniques with the aid of immunohistochemistry and/or immunofluorescence methods. In 1-week-old neonatal testis, Crlz-1 was strongly expressed in the spermatogonia and Sertoli cells in its seminiferous cord. In 2- to 3-week-old prepubertal testis, where Sertoli cells cease to proliferate, Crlz-1 expression dropped and remained weakly at the rim layer of seminiferous cords and/or tubules, where spermatogonia are present. In the adult testis at 12 weeks after birth, Crlz-1 was expressed mainly in the spermatids near the lumen of seminiferous tubules. In a further in situ hybridization of Crlz-1 in the 12-week-old adult testis with hematoxylin nuclear counterstaining, Crlz-1 was mainly expressed at step 16 of spermatids between stages VII and VIII of seminiferous tubules as well as in their residual bodies at stage IX of seminiferous tubules. PMID:23525569

Lim, Jung-Hyun; Choi, Seong-Young; Yoo, Han-Woong; Cho, Sun-Jung; Son, Youngsook; Kang, Chang-Joong

2013-07-01

152

Large cell calcifying sertoli cell tumor of the testis: a case study and review of the literature.  

PubMed

A 24-year-old man was admitted due to an incidentally detected mass in his left testis, which showed radiopaque calcification on plain X-ray film. Left orchiectomy was performed, and the resected testis contained a well-demarcated, hard mass measuring 1.1 cm. Histological analysis revealed that the tumor was composed of neoplastic cells, fibrotic stroma, and laminated or irregularly shaped calcific bodies. The individual cells had abundant eosinophilic or clear cytoplasm with round nuclei, each of which contained one or two conspicuous nucleoli. They were arranged in cords, trabeculae, clusters, and diffuse sheets. There were several foci of intra-tubular growth patterns, with thickening of the basal lamina. Immunohistochemically, the neoplastic cells were positive for S-100 protein and vimentin, focally positive for inhibin alpha, and negative for cytokeratin, CD10, and Melan-A. In addition to reporting this rare case, we also review the relevant literature regarding large cell calcifying Sertoli cell tumors. PMID:24627695

Song, Dae Hyun; Jeong, Seong Muk; Park, Jong Tak; Yun, Gak Won; Kim, Byoung Kwon; Lee, Jong Sil

2014-02-01

153

The Dynamic of the Apical Ectoplasmic Specialization between Spermatids and Sertoli Cells: The Case of the Small GTPase Rap1  

PubMed Central

Despite advances in assisted reproductive technologies, infertility remains a consistent health problem worldwide. Spermiation is the process through which mature spermatids detach from the supporting Sertoli cells and are released into the tubule lumen. Spermiation failure leads to lack of mature spermatozoa and, if not occasional, could result into azoospermia, major cause of male infertility in human population. Spermatids are led through their differentiation into spermatozoa by the apical ectoplasmic specialization (aES), a testis-specific, actin-based anchoring junction restricted to the Sertoli-spermatid interface. The aES helps spermatid movement across the seminiferous epithelium, promotes spermatid positioning, and prevents the release of immature spermatozoa. To accomplish its functions, aES needs to undergo tightly and timely regulated restructuring. Even if components of aES are partly known, the mechanism/s through which aES is regulated remains still elusive. In this review, we propose a model by which the small GTPase Rap1 could regulate aES assembly/remodelling. The characterization of key players in the dynamic of aES, such as Rap1, could open new possibility to develop prognostic, diagnostic, and therapeutic approaches for male patients under treatment for infertility as well as it could lead to the identification of new target for male contraception.

2014-01-01

154

The dynamic of the apical ectoplasmic specialization between spermatids and Sertoli cells: the case of the small GTPase Rap1.  

PubMed

Despite advances in assisted reproductive technologies, infertility remains a consistent health problem worldwide. Spermiation is the process through which mature spermatids detach from the supporting Sertoli cells and are released into the tubule lumen. Spermiation failure leads to lack of mature spermatozoa and, if not occasional, could result into azoospermia, major cause of male infertility in human population. Spermatids are led through their differentiation into spermatozoa by the apical ectoplasmic specialization (aES), a testis-specific, actin-based anchoring junction restricted to the Sertoli-spermatid interface. The aES helps spermatid movement across the seminiferous epithelium, promotes spermatid positioning, and prevents the release of immature spermatozoa. To accomplish its functions, aES needs to undergo tightly and timely regulated restructuring. Even if components of aES are partly known, the mechanism/s through which aES is regulated remains still elusive. In this review, we propose a model by which the small GTPase Rap1 could regulate aES assembly/remodelling. The characterization of key players in the dynamic of aES, such as Rap1, could open new possibility to develop prognostic, diagnostic, and therapeutic approaches for male patients under treatment for infertility as well as it could lead to the identification of new target for male contraception. PMID:24719879

Berruti, Giovanna; Paiardi, Chiara

2014-01-01

155

Sertoli cells in culture secrete paracrine factor(s) that inhibit peritubular myoid cell proliferation: identification of heparinoids as likely candidates  

SciTech Connect

Conditioned medium from Sertoli cells, prepared from testes of 20-day-old rats, contains component(s) that inhibit the incorporation of (3H)-thymidine into DNA of peritubular myoid cells (PMC) and inhibit the proliferation of PMC. These components are trypsin-resistant, heat-stable compounds having a molecular weight less than 30,000. The active inhibitory components in Sertoli cell conditioned medium are inactivated by treatment with heparinase, but not by treatment with hyaluronidase or chondroitin sulfate lyases. Addition of heparin or heparan sulfate results in inhibition of DNA synthesis by PMC in a dose-dependent manner, whereas other glycosaminoglycans (GAGs) examined (hyaluronic acid, keratan sulfate, and chondroitin sulfate) have no detectable effects. Heparin and heparan sulfate are unique among GAGs tested in inhibiting the characteristic multilayer growth pattern of PMC following the attainment of confluence in serum-rich medium. On the basis of these and other data presented, it is concluded that heparin and other heparin-like GAGs synthesized by Sertoli cells are implicated in the modulation of growth of PMC in vitro during co-culture. It is postulated that heparin may play a similar role in maintaining the quiescent peritubular myoid cell phenotype in vivo.

Tung, P.S.; Fritz, I.B. (Banting and Best Department of Medical Research, University of Toronto, Ontario (Canada))

1991-06-01

156

Protective role of lycopene on polychlorinated biphenyls (Aroclor 1254)-induced adult rat Sertoli cell dysfunction by increased oxidative stress and endocrine disruption  

Microsoft Academic Search

The protective role of lycopene against various toxicants on various organs and its association with decreased oxidative stress seems to be well established. Sertoli cell function in the adult testis determines daily sperm production. Increasing evidence suggests that environmental pollutants including polychlorinated biphenyls (PCBs) are male infertility, which is associated with oxidative stress. Hence, the present study was designed to

G. Krishnamoorthy; K. Selvakumar; P. Elumalai; P. Venkataraman; J. Arunakaran

2011-01-01

157

Expression of inhibin-alpha is regulated synergistically by Wilms' tumor gene 1 (Wt1) and steroidogenic factor-1 (Sf1) in sertoli cells.  

PubMed

Wt1 encodes a zinc finger nuclear transcriptional factor, which is specifically expressed in testicular Sertoli cells and knockdown of Wt1 in Sertoli cells causes male mice subfertility. However, the underlying mechanism is still unclear. In this study, we found that expression of inhibin-? is significantly reduced in Wt1-deficient Sertoli cells. Luciferase assays using the inhibin-? promoter indicated that the inhibin-? promoter is transactivated by the Wt1 A, and B isoforms (-KTS), but not the C, and D isoforms (+KTS). Analysis of the Wt1 responsive element of the inhibin-? promoter region using site-directed mutagenesis showed that the nucleotides between -58 and -49 are essential for Wt1-dependent transactivation of the inhibin-? promoter. ChIP assays indicated that Wt1 directly interacts with the inhibin-? promoter. In addition, the inhibin-? promoter is activated synergistically by Wt1 and Sf1. Mutation of the ligand binding domain (LBD) of Sf1 (residues 235-238) completely abolished the synergistic action between Wt1 and Sf1, but did not affect the physical interaction between these two proteins, suggesting that other factor(s) may also be involved in the regulation of inhibin-? in Sertoli cells. Further studies demonstrated that ?-catenin enhances the synergistic activation of Wt1 and Sf1 on the inhibin-? promoter. Given the fact that inhibin-?, a subunit of inhibin, is known to be involved in the regulation of spermatogenesis and testicular steroidogenesis, this study reveals a new regulatory mechanism of inhibin-? in Sertoli cells and also sheds light on the physiological functions of Wt1 in gonad development and spermatogenesis. PMID:23326390

Ji, Shao-Yang; Hao, Jian-Xiu; Li, Lei; Zhang, Jun; Zheng, Qiao-Song; Li, Xi-Xia; Wang, Xiao-Na; Han, Chun-Sheng; Gao, Fei; Liu, Yi-Xun

2013-01-01

158

FSH-induced phosphoprotein phosphatase 2A-mediated deactivation of particulate phosphodiesterase-4 activities is abolished after alteration in proteoglycan synthesis in immature rat Sertoli cells.  

PubMed

Cessation of rat testicular Sertoli cells proliferation around days 15-20 post partum is associated in vitro with the highest rise in rolipram-sensitive cAMP-catabolizing phosphodiesterase-4 activities (PDE4s) triggered by FSH during the early postnatal period. The transient nature of FSH-induced increase in PDE4s suggests concomitant changes in both PKA-mediated activation and subsequent deactivation of these activities. In this study, we demonstrated that the deactivation of FSH-stimulated particulate, but not soluble, PDE4s in cultured Sertoli cells from 20-day-old rats was inhibited by phosphoprotein phosphatase (PP) inhibitors, okadaïc acid, and calyculin A. Moreover, the deactivation of FSH-stimulated particulate PDE4s was timely related with the gonadotropin-induced increase in both particulate PP2A activity and particulate PP2A catalytic subunit immunoreactive expression independently of any transcriptional regulation of that subunit. Both the FSH-induced increase in recruitment/activation of particulate PP2A and the subsequent deactivation of particulate PDE4 were abolished when Sertoli cell proteoglycans (PGs) synthesis was altered by sodium chlorate. Sodium chlorate effect was developmentally regulated as evidenced by its ability to silence particulate PDE4 deactivation only in non-proliferating (from 20- to 30-day-old rats) but not in proliferating (from 10-day-old rats) Sertoli cells. All these data suggested that PGs could be involved in the FSH-induced recruitment/activation of PP2A. Particularly, developmentally regulated transmembrane syndecans, the most abundant PGs in Sertoli cells, by targeting PP2A at the membrane level could allow developmental control of activated particulate PDE4s and, potentially, other signaling phosphoproteins, including the FSH receptor, during the early postnatal period. PMID:18372231

Levallet, Guénaëlle; Levallet, Jérôme; Bonnamy, Pierre-Jacques

2008-04-01

159

The PGD2 pathway, independently of FGF9, amplifies SOX9 activity in Sertoli cells during male sexual differentiation  

PubMed Central

Activation by the Y-encoded testis determining factor SRY and maintenance of expression of the Sox9 gene encoding the central transcription factor of Sertoli cell differentiation are key events in the mammalian sexual differentiation program. In the mouse XY gonad, SOX9 upregulates Fgf9, which initiates a Sox9/Fgf9 feedforward loop, and Sox9 expression is stimulated by the prostaglandin D2 (PGD2) producing lipocalin prostaglandin D synthase (L-PGDS, or PTDGS) enzyme, which accelerates commitment to the male pathway. In an attempt to decipher the genetic relationships between Sox9 and the L-Pgds/PGD2 pathway during mouse testicular organogenesis, we found that ablation of Sox9 at the onset or during the time window of expression in embryonic Sertoli cells abolished L-Pgds transcription. By contrast, L-Pgds-/- XY embryonic gonads displayed a reduced level of Sox9 transcript and aberrant SOX9 protein subcellular localization. In this study, we demonstrated genetically that the L-Pgds/PGD2 pathway acts as a second amplification loop of Sox9 expression. Moreover, examination of Fgf9-/- and L-Pgds-/- XY embryonic gonads demonstrated that the two Sox9 gene activity amplifying pathways work independently. These data suggest that, once activated and maintained by SOX9, production of testicular L-PGDS leads to the accumulation of PGD2, which in turn activates Sox9 transcription and nuclear translocation of SOX9. This mechanism participates together with FGF9 as an amplification system of Sox9 gene expression and activity during mammalian testicular organogenesis.

Moniot, Brigitte; Declosmenil, Faustine; Barrionuevo, Francisco; Scherer, Gerd; Aritake, Kosuke; Malki, Safia; Marzi, Laetitia; Cohen-Solal, Anne; Georg, Ina; Klattig, Jurgen; Englert, Christoph; Kim, Yuna; Capel, Blanche; Eguchi, Naomi; Urade, Yoshihiro; Boizet-Bonhoure, Brigitte; Poulat, Francis

2009-01-01

160

17beta-estradiol induces the translocation of the estrogen receptors ESR1 and ESR2 to the cell membrane, MAPK3/1 phosphorylation and proliferation of cultured immature rat Sertoli cells.  

PubMed

The aim of the present study was to determine the mechanisms involved in estrogen actions in cultured rat Sertoli cells. RT-PCR detected transcripts for the estrogen receptors ESR1 and ESR2 in cultured immature Sertoli cells and in the testis of 15-, 28-, and 120-day-old rats. The expression of ESR1 and ESR2 was confirmed in Sertoli cells by immunofluorescence and Western blot. Immunohistochemistry with cryosections of testes from immature and adult rats revealed that ESR1 is present in Sertoli, Leydig, and some peritubular myoid cells, and ESR2 is present in multiple cell types, including germ cells. Treatment of Sertoli cells with 17beta-estradiol (E(2)) induced a translocation of ESR1 and ESR2 to the plasma membrane and a concomitant phosphorylation of MAPK3/1. Both effects reached a maximum after 10 min and were blocked by PP2, an inhibitor of the SRC family of protein tyrosine kinases, and by the antiestrogen ICI 182,780 (ICI). MAPK3/1 phosphorylation was also decreased in the presence of AG 1478, an inhibitor of the epidermal growth factor receptor (EGFR) kinase, and in the presence of MAP2K1/2 inhibitor UO126. Treatment with E(2) for 24 h increased the incorporation of [methyl-(3)H]thymidine, which was blocked by ICI. These results indicate that E(2) activates an SRC-mediated translocation of estrogen receptors to the plasma membrane, which results in the activation of EGFR and the mitogen-activated protein kinase signaling pathway. In addition, activation of ESR1 and/or ESR2 by E(2) is involved in proliferation of immature Sertoli cells. The estrogen actions in Sertoli cells might be a key step mediating cellular events important for spermatogenesis and fertility. PMID:17928626

Lucas, Thaís F G; Siu, Erica R; Esteves, Carlos A; Monteiro, Hugo P; Oliveira, Cleida A; Porto, Catarina S; Lazari, Maria Fatima M

2008-01-01

161

Nonylphenol-induced apoptotic cell death in mouse TM4 Sertoli cells via the generation of reactive oxygen species and activation of the ERK signaling pathway.  

PubMed

Nonylphenol (NP), a representative endocrine disruptor, interferes with reproductive function in aquatic organisms and animals. Although many previous studies have focused on apoptotic cell death by NP, the fundamental mechanism of NP on apoptosis remains poorly understood. Here, we investigated the molecular mechanism on NP-induced apoptotic cell death in mouse TM4 Sertoli cells. To evaluate NP treatment on cell viability, formazan and lactate dehydrogenase (LDH) assays were performed. Results indicate that NP reduced cell viability and increased the release of LDH in dose- and time-dependent manners. The reduction of cell viability by NP treatment appeared to involve necrosis as well as apoptosis based on nuclear fragmentation, an increase in the sub G1 population, and the detection of poly(ADP ribose) polymerase and caspase-3 cleavage. Additionally, the anti-apoptotic protein Bcl-2 diminished, whereas the pro-apoptotic protein Bax increased in a time-dependent manner. Note that NP-induced apoptotic cell death was enhanced by the generation of reactive oxygen species (ROS) and activation of extracellular signal-regulated kinase (ERK) signaling. Pretreatment with N-acetylcysteine, an antioxidant, attenuated NP-induced apoptotic cell death. Moreover, NP caused a transient activation of the MAPK pathway. In particular, NP-induced cell death was significantly suppressed by U0126, a specific inhibitor of ERK. Taken together, our results suggest that NP induces apoptosis in mouse TM4 Sertoli cells via ROS generation and ERK activation. Copyright © 2013 John Wiley & Sons, Ltd. PMID:23677851

Choi, Mi-Sun; Park, Han-Jin; Oh, Jung-Hwa; Lee, Eun-Hee; Park, Se-Myo; Yoon, Seokjoo

2014-06-01

162

Sertoli cell cycle: a re-examination of the structural changes during the cycle of the seminiferous epithelium of the rat.  

PubMed

Seminiferous tubules in mammals are composed of cell associations that show a cyclic pattern of renewal and development. The cyclic nature of germ cell development suggests that the cells supporting the spermatogenic process, the Sertoli cells, might also differ structurally during the spermatogenic cycle in terms of the quantity of their constituents. In the present study, cyclic differences in volumes, and surface areas were determined using a sampling technique at the electron microscope level that proportionally samples the Sertoli cell within the seminiferous tubule. Among the many parameters studied, only the surface area of the cell, the volume of lipid, and the volume and surface area of the rough endoplasm reticulum were shown by statistical analysis to vary cyclically. Regarding rough endoplasm reticulum, the volume and surface area of this organelle peaked at mid-cycle and its low was recorded near the end of the cycle, exhibiting an approximate 15-fold difference between extremes. The rough endoplasm reticulum parameters generally correlated with known patterns of protein secretion within the tubule and with the secretion of specific proteins as well as the factors important in controlling protein secretion. Many Sertoli cell structural parameters suggested to be influenced cyclically in the rat in other studies could not be confirmed by the present study. Methodological differences in the present study and past studies are discussed as potential sources of error for these discrepancies. PMID:8238970

Ye, S J; Ying, L; Ghosh, S; de França, L R; Russell, L D

1993-10-01

163

Ultrastructure of germ cells, Sertoli cells and mitochondria during spermatogenesis in mature testis of the Chinese Taihang black goats (Capra hircus).  

PubMed

The objective of this study was to describe the ultrastructure of germ cells, Sertoli cells and mitochondria in mature testis of the Chinese Taihang black goat. The characteristics of germ cell nucleus and mitochondria changing during spermatogenesis were investigated by transmission electron microscopy (TEM). The results showed that the spermatogonium was elliptical, and its nucleus was about 4-5 ?m. The round mitochondria can be observed throughout the cytoplasm around the nucleus. Small patches of heterochromatin were distributed throughout the nucleus. Spermatocyte was oval-shaped with a nucleus of about 4-4.5 ?m in diameter. The heterochromatin began to attach to the inner surface of the nuclear membrane. Spermatid was about 4 ?m and oval in shape. Its nucleus was oval or round and approximately 2-3 ?m in diameter. The borderline between nucleus membrane and karyoplasm was distinct. During spermiogenesis, spermatid nucleus was condensed and elongated, and chromatin reached the highest condensation in the mature spermatozoon. The mid-piece was surrounded by mitochondria at the neck region. The sperm tail showed the typical "9+2? structure, contained axoneme and central singlet microtubules. The nuclei of the Sertoli cells were irregular shaped and showed indentations in the membrane. In the mature testes of goat bucks, abundant mitochondria were around the germ cells and Sertoli cells. The scattered mitochondria were aggregated around the base of the flagellum (axoneme) during the spermatid differentiation stage. In conclusion, the present study showed that the spermatogenic process of Taihang black goat followed the pattern of mammals with some specific. PMID:23618746

Shi, Liguang; Xun, Wenjuan; Zhou, Hanlin; Hou, Guanyu; Yue, Wenbin; Zhang, Chunxiang; Ren, Youshe; Yang, Rujie

2013-07-01

164

The effects of di(2-ethylhexyl)phthalate exposure and selenium nutrition on sertoli cell vimentin structure and germ-cell apoptosis in rat testis.  

PubMed

This study aimed to investigate the effects of di(2-ethylhexyl)phthalate (DEHP) on Sertoli-cell vimentin filaments and germ-cell apoptosis in testes of pubertal rats at different selenium (Se) status. Se deficiency was produced in 3-weeks old Sprague-Dawley rats by feeding them ? 0.05 Se mg/kg diet for 5 weeks, Se supplementation group was on 1 mg Se/kg diet, and DEHP was applied at 1000 mg/kg dose by gavage during the last 10 days of the feeding period. The diet with excess Se did not cause any appreciable alteration in vimentin staining and apoptosis of germ cells, but Se deficiency caused a mild decrease in the intensity of vimentin immunoreactivity and enhanced germ-cell apoptosis significantly (approximately 3-fold, p <0.0033). DEHP exposure caused disruption and collapse of vimentin filaments and significantly induced apoptotic death of germ cells (approximately 8-fold, p <0.0033). In DEHP-exposed Se-deficient animals, compared with the control, collapse of vimentin filaments was more prominent; there was serious damage to the seminiferous epithelium; and a high increment (approximately 25-fold, p <0.0033) in apoptotic germ cells was observed. Thus, Se deficiency exacerbated the toxicity of DEHP on Sertoli cells and spermatogenesis, whereas Se supplementation provided protection. These results put forward the critical role of Se in the modulation of redox status of testicular cells and emphasize the importance of Se status for reproductive health. PMID:22002783

Erkekoglu, Pinar; Zeybek, N Dilara; Giray, Belma; Asan, Esin; Hincal, Filiz

2012-04-01

165

Implication of actin microfilaments in maintenance of intercellular bridges and the Sertoli cell barrier in the rat seminiferous epithelium  

SciTech Connect

Within the seminiferous epithelium, germ cells are connected to one another by intercellular bridges. Additionally, young germ cells are separated from more advanced germ cells by the Sertoli cell barrier, the occluding junctions of which are associated with actin microfilaments. To examine how microfilaments influence these structures in the rat the actin-disrupting agent cytochalasin D (CD) was injected intratesticularly (i.t.). In preliminary studies the optical injection volume was found to be 50 {mu}l and, by using the dye trypan blue, the injected solution was shown to enter the lymphatic system and rapidly spread throughout the testis. A 50% clearance of {sup 3}H-insulin from the testis was achieved at 3 hr and 95% by 24 hr. Vehicles with varying solubility properties did not cause testicular damage. Intercellular bridges were found to be dynamic structures. As spermatogenesis progressed, the bridge diameter gradually increased. The formation and degradation of bridge partitioning complexes within pre-existing bridges of dividing cells were described.

Weber, J.E.

1987-01-01

166

Cytokines, Polarity Proteins, and Endosomal Protein Trafficking and Signaling--The Sertoli Cell Blood-Testis Barrier System In Vitro as a Study Model  

PubMed Central

Endosomal signaling is emerging as one of the most important cellular events that regulate signaling function in mammalian cells or an epithelium in response to changes in environment such as the presence of stimuli mediated by cytokines, toxicants, heat, ions during growth and development, and other cellular processes such as cytokinesis and spermatogenesis. Recent studies have shown that protein endocytosis—the initial step of endosomal signaling—involves the participation of polarity proteins, such as partitioning defective protein 6 (Par6), Cdc42 and 14-3-3 (also known as Par5), which in turn is regulated by cytokines (e.g., TGF-?2, TGF-?3) and testosterone at the Sertoli cell blood–testis barrier (BTB) in the mammalian testis. In this short method paper, we provide a detailed protocol of assessing protein endocytosis, the initial and also the most critical step of endosomal signaling at the Sertoli cell BTB. This biochemical endocytosis assay summarizes our experience for the last decade, which should likely be performed in conjunction with the dual-labeled immunofluorescence analysis to assess protein endocytosis. While we are using a Sertoli cell in vitro system that mimics the BTB in vivo, this approach should be applicable to virtually all mammalian cells.

Xiao, Xiang; Wong, Elissa W.P.; Lie, Pearl P.Y.; Mruk, Dolores D.; Wong, Chris K.C.; Cheng, C. Yan

2014-01-01

167

Human Leydig Cells and Sertoli Cells Are Producers of Interleukins1 and -6  

Microsoft Academic Search

Interleukins (IL)-1 and -6 have been shown to be produced by several categories of cells in the rat testis and involved in the paracrine control of testicular function. Evidence of high amounts of IL-1 have been shown in the human testis, but nothing is known about its cellular origin. Furthermore, to our knowledge, the presence of IL-6 in the human

CORINNE CUDICINI; EDITH GOMEZ; EUGENE BOSMANS; FRANCOIS BALLET; BERNARD JEGOU

2010-01-01

168

Effects of Vitamin C and E on PCB (Aroclor 1254) induced oxidative stress, androgen binding protein and lactate in rat Sertoli cells  

Microsoft Academic Search

The effect of Aroclor 1254 and the ameliorative effect of Vitamin C and E on Sertoli cell function were studied in adult male rats. The rats were administered Aroclor 1254 at a dose of 2mg\\/kg bw\\/day intraperitoneally for 30 days. One group of rats received Vitamin C (100mg\\/kg bw\\/day) while the other group received Vitamin E (50mg\\/kg bw\\/day) orally simultaneously

J. Senthil kumar; S. Banudevi; M. Sharmila; P. Murugesan; N. Srinivasan; K. Balasubramanian; M. M. Aruldhas; J. Arunakaran

2004-01-01

169

Follicle-stimulating hormone regulates the expression of cyclic protein-2\\/cathepsin L messenger ribonucleic acid in rat Sertoli cells in a stage-specific manner  

Microsoft Academic Search

Cyclic protein-2\\/cathepsin L (CP-2) is secreted by Sertoli cells in a highly stage-specific manner, maximally during stages VI–VII of the rat seminiferous epithelial cycle. We investigated FSH regulation of CP-2 mRNA expression and its cellular localization in isolated staged seminiferous tubular segments. FSH induced a significant increase of CP-2 mRNA levels in stages IX-I, whereas in stages II–VIII, the levels

Tarja-Leena Penttilä; Harri Hakovirta; Pekka Mali; William W. Wright; Martti Parvinen

1995-01-01

170

Retinol increases catalase activity and protein content by a reactive species-dependent mechanism in Sertoli cells.  

PubMed

Vitamin A (retinol) is widely used as an antioxidant in therapeutic interventions and dietary supplementations. However, the redox properties of retinoids have been the subject of intense debate in the last few years, as recent works observed deleterious effects caused by retinol supplementation in clinical trials. In the present work, we show that retinol treatment (7 microM, 24 h) led to catalase (EC 1.11.1.6; CAT) activation in cultured Sertoli cells by increasing its protein content in a reactive species-dependent manner. Retinol treatment also increased cell lipoperoxidation, assessed by determination of thiobarbituric acid-reactive substances (TBARS), and intracellular reactive species production, determined by the real-time dihydrochlorofluorescein (DCFH-DA) assay. However, no alterations on CAT mRNA expression (assessed by RT-PCR) were observed, indicating an effect independent of CAT gene-transcription regulation. Importantly, all the effects induced by retinol were inhibited by the antioxidant Trolox, a hydrophilic analogue of alpha-tocopherol. These results show for the first time that retinol increases CAT activity by a redox-dependent modulation of its protein content in a cell culture model. CAT activity or expression are widely used as indexes of oxidative stress in biological systems; since no changes in CAT mRNA expression were detected in these conditions, the use of CAT gene-transcription activation when assessing oxidative stress should be re-evaluated. PMID:18533141

Gelain, Daniel Pens; de Bittencourt Pasquali, Matheus Augusto; Zanotto-Filho, Alfeu; de Souza, Luiz Fernando; de Oliveira, Ramatis Birnfeld; Klamt, Fábio; Moreira, José Cláudio Fonseca

2008-07-10

171

Sertoli cell androgen receptor expression regulates temporal fetal and adult Leydig cell differentiation, function, and population size.  

PubMed

We recently created a mouse model displaying precocious Sertoli cell (SC) and spermatogenic development induced by SC-specific transgenic androgen receptor expression (TgSCAR). Here we reveal that TgSCAR regulates the development, function, and absolute number of Leydig cells (LCs). Total fetal and adult type LC numbers were reduced in postnatal and adult TgSCAR vs control testes, despite normal circulating LH levels. Normal LC to SC ratios found in TgSCAR testes indicate that SC androgen receptor (SCAR)-mediated activity confers a quorum-dependent relationship between total SC and LC numbers. TgSCAR enhanced LC differentiation, shown by elevated ratios of advanced to immature LC types, and reduced LC proliferation in postnatal TgSCAR vs control testes. Postnatal TgSCAR testes displayed up-regulated expression of coupled ligand-receptor transcripts (Amh-Amhr2, Dhh-Ptch1, Pdgfa-Pdgfra) for potential SCAR-stimulated paracrine pathways, which may coordinate LC differentiation. Neonatal TgSCAR testes displayed normal T and dihydrotestosterone levels despite differential changes to steroidogenic gene expression, with down-regulated Star, Cyp11a1, and Cyp17a1 expression contrasting with up-regulated Hsd3b1, Hsd17b3, and Srd5a1 expression. TgSCAR males also displayed elevated postnatal and normal adult serum testosterone levels, despite reduced LC numbers. Enhanced adult-type LC steroidogenic output was revealed by increased pubertal testicular T, dihydrotestosterone, 3?-diol and 3?-diol levels per LC and up-regulated steroidogenic gene (Nr5a1, Lhr, Cyp11a1, Cyp17a1, Hsd3b6, Srd5a1) expression in pubertal or adult TgSCAR vs control males, suggesting regulatory mechanisms maintain androgen levels independently of absolute LC numbers. Our unique gain-of-function TgSCAR model has revealed that SCAR activity controls temporal LC differentiation, steroidogenic function, and population size. PMID:23766127

Hazra, Rasmani; Jimenez, Mark; Desai, Reena; Handelsman, David J; Allan, Charles M

2013-09-01

172

Activity and form of sulfated glycoprotein 2 (clusterin) from cultured Sertoli cells, testis, and epididymis of the rat.  

PubMed

Sulfated glycoprotein 2 (SGP 2) is a 73-kDa highly glycosylated disulfide-linked heterodimer. It is a major secreted protein of Sertoli cells, is found in high abundance within the seminiferous tubule fluid (STF) and epididymal fluid (EPF), and can be found on the surface of spermatozoa. Due to its high abundance and location it is believed to play a major role in the development of spermatozoa; however, its specific function(s) within the reproductive tract is not known. Purified and renatured SGP 2 were found to have the ability to inhibit complement activity with a mean concentration of 66 mg/ml for 50% inhibition. Extraction of epididymal sperm with various reagents showed that a major fraction of the SGP 2 in EPF was free or was loosely associated with the spermatozoa whereas a smaller fraction was more tightly associated and disruption of the lipid bilayer was required for its complete removal. Ultracentrifugation techniques and gel permeation chromatography revealed that SGP 2 in plasma, STF, and EPF formed complexes with other proteins and/or lipids but was not specifically associated with the apolipoprotein-like particles containing apolipoprotein A1 (apo A1). PMID:8167239

Law, G L; Griswold, M D

1994-03-01

173

Retinol and retinoic acid modulate catalase activity in Sertoli cells by distinct and gene expression-independent mechanisms.  

PubMed

Vitamin A (retinol) exerts a major role in several biological functions. However, it was observed that retinol induces oxidative stress on different cellular types. Catalase (EC 1.11.1.6; CAT) is a hydrogen peroxide metabolizing enzyme, and its activity and expression is widely used as an index to measure oxidative stress and perturbations in the cellular redox state. The aim of this study was to investigate the effects of retinol and its major biologically active metabolite, all-trans retinoic acid (RA), on CAT regulation. For this purpose, cultured Sertoli cells (a physiological target of vitamin A) were treated with retinol or RA. Retinol (7 microM, 14 microM) and RA (100 nM, 1 microM) enhanced intracellular reactive species production and increased CAT activity after 24 h of treatment. Retinol increased CAT immunocontent but did not alter CAT mRNA expression, while the increase in CAT activity by RA was not related to alterations in immunocontent or mRNA expression. In vitro incubation of purified CAT with retinol or RA did not alter enzyme activity. PMID:18440196

Pasquali, Matheus A B; Gelain, Daniel P; Zanotto-Filho, Alfeu; de Souza, Luiz F; de Oliveira, Ramatis B; Klamt, Fabio; Moreira, Jose C F

2008-08-01

174

Melatonin regulates the development and function of bovine Sertoli cells via its receptors MT1 and MT2.  

PubMed

Melatonin and its receptors are found in the testis of many species, where they mediate testicular functions. The present study aimed to investigate the expression of melatonin receptors (MT1 and MT2) in bovine Sertoli cells (SCs), using reverse transcription polymerase chain reaction (RT-PCR) and western blot. In addition, we assessed the mRNA levels of spermatogenesis-related genes (real-time PCR) and secretion of inhibin B after treatment with various concentrations (0, 80, 160, and 320 pg/mL) of melatonin at different time points (24, 48, or 72 h). We found that bovine SCs express MT1 and MT2 receptors, which were regulated by melatonin in time- and dose-dependent manners after treatment with melatonin. Exogenous melatonin up-regulated the expression of spermatogenesis-related genes, including Cyclin D1, Cyclin E, Pdgfa, Dhh, Occludin, and Claudin, and decreased the mRNA levels of P21 and Kit1 in a time or dose-dependent manner. Meanwhile, melatonin supplementation significantly affected Inhba, Inhbb and Inha mRNA expression. These findings were consistent with inhibin B levels detected in the culture medium. In conclusion, exogenous melatonin acts via its receptors and appears to play regulatory roles in the development and function of bovine SCs. PMID:24768045

Yang, Wu-Cai; Tang, Ke-Qiong; Fu, Chang-Zhen; Riaz, Hasan; Zhang, Qiong; Zan, Lin-Sen

2014-06-10

175

B Cells Regulate Murine Gammaherpesvirus 68 Latency  

Microsoft Academic Search

The dynamics of the establishment of, and reactivation from, gammaherpesviruses latency has not been quantitatively analyzed in the natural host. Gammaherpesvirus 68 (gHV68) is a murine gammaherpesvirus genetically related to primate gammaherpesviruses that establishes a latent infection in infected mice. We used limiting dilution reactivation (frequency of cells reactivating gHV68 in vitro) and limiting dilution PCR (frequency of cells carrying

KAREN E. WECK; SUSANNE S. KIM; HERBERT W. VIRGIN; SAMUEL H. SPECK

1999-01-01

176

Sertoli-cell-specific knockout of connexin 43 leads to multiple alterations in testicular gene expression in prepubertal mice  

PubMed Central

SUMMARY A significant decline in human male reproductive function has been reported for the past 20 years but the molecular mechanisms remain poorly understood. However, recent studies showed that the gap junction protein connexin-43 (CX43; also known as GJA1) might be involved. CX43 is the predominant testicular connexin (CX) in most species, including in humans. Alterations of its expression are associated with different forms of spermatogenic disorders and infertility. Men with impaired spermatogenesis often exhibit a reduction or loss of CX43 expression in germ cells (GCs) and Sertoli cells (SCs). Adult male transgenic mice with a conditional knockout (KO) of the Gja1 gene [referred to here as connexin-43 (Cx43)] in SCs (SCCx43KO) show a comparable testicular phenotype to humans and are infertile. To detect possible signaling pathways and molecular mechanisms leading to the testicular phenotype in adult SCCx43KO mice and to their failure to initiate spermatogenesis, the testicular gene expression of 8-day-old SCCx43KO and wild-type (WT) mice was compared. Microarray analysis revealed that 658 genes were significantly regulated in testes of SCCx43KO mice. Of these genes, 135 were upregulated, whereas 523 genes were downregulated. For selected genes the results of the microarray analysis were confirmed using quantitative real-time PCR and immunostaining. The majority of the downregulated genes are GC-specific and are essential for mitotic and meiotic progression of spermatogenesis, including Stra8, Dazl and members of the DM (dsx and map-3) gene family. Other altered genes can be associated with transcription, metabolism, cell migration and cytoskeleton organization. Our data show that deletion of Cx43 in SCs leads to multiple alterations of gene expression in prepubertal mice and primarily affects GCs. The candidate genes could represent helpful markers for investigators exploring human testicular biopsies from patients showing corresponding spermatogenic deficiencies and for studying the molecular mechanisms of human male sterility.

Giese, Sarah; Hossain, Hamid; Markmann, Melanie; Chakraborty, Trinad; Tchatalbachev, Svetlin; Guillou, Florian; Bergmann, Martin; Failing, Klaus; Weider, Karola; Brehm, Ralph

2012-01-01

177

Morphological evidences indicate that the interference of cimetidine on the peritubular components is responsible for detachment and apoptosis of Sertoli cells  

PubMed Central

Cimetidine, referred as antiandrogenic agent, has caused alterations in the seminiferous tubules, including alterations in the peritubular tissue and death of myoid cells by apoptosis. Regarding the structural and functional importance of the peritubular tissue for the maintenance of Sertoli cells (SC), we purpose to investigate the SC-basement membrane interface, focusing the morphological features of SC and their interaction with the basement membrane in the affected tubules by cimetidine. Ten animals were distributed into two groups, control (CG) and cimetidine (CmG) which received saline solution and 50 mg of cimetidine per kg of body weight, respectively, for 52 days. The testes were fixed, dehydrated and embedded for analyses under light and transmission electron microscopy. Paraffin sections were submitted to the TUNEL method; sections of testes embedded in glycol methacrylate were submitted to PAS method and stained by H&E for morphological and quantitative analyses of Sertoli Cells. In the CmG, the SC nuclei were positive to the TUNEL method and showed typical morphological alterations of cell death by apoptosis (from early to advanced stages). A significant reduction in the number of Sertoli Cells was probably due to death of these cells by apoptosis. A close relationship between SC nuclear alterations (including a high frequency of dislocated nuclei from the basal portion) and damage in the peritubular tissue was observed. The ultrastructural analysis showed a parallelism between the gradual advancement of apoptotic process in SC and detachment of the anchoring sites (hemidesmosomes) of SC plasma membrane from the lamina densa. The presence of portions of lamina densa underlying the detached hemidesmosomes indicates a continuous deposition of lamina densa, resulting in the thickening of the basal lamina. The results indicate a possible disarrangement of the SC cytoskeleton, including the focal adhesion structure. These alterations are related to SC apoptosis and probably result from disturbs induced by cimetidine on the peritubular tissue.

Sasso-Cerri, Estela; Cerri, Paulo S

2008-01-01

178

Blood-testis barrier dynamics are regulated by testosterone and cytokines via their differential effects on the kinetics of protein endocytosis and recycling in Sertoli cells.  

PubMed

During spermatogenesis in the mammalian testis, preleptotene/leptotene spermatocytes differentiate from type B spermatogonia and traverse the blood-testis barrier (BTB) at stage VIII of the seminiferous epithelial cycle for further development. This timely movement of germ cells involves extensive junction restructuring at the BTB. Previous studies have shown that these events are regulated by testosterone (T) and cytokines [e.g., the transforming growth factor (TGF) -betas], which promote and disrupt the BTB assembly, respectively. However, the mechanisms underlying the "opening" of the BTB above a migrating preleptotene/leptotene spermatocyte and the "resealing" of the barrier underneath this cell remain obscure. We now report findings on a novel mechanism utilized by the testes to regulate these events. Using cell surface protein biotinylation coupled with immunoblotting and immunofluorescent microscopy, we assessed the kinetics of endocytosis and recycling of BTB-associated integral membrane proteins: occludin, JAM-A, and N-cadherin. It was shown that these proteins were continuously endocytosed and recycled back to the Sertoli cell surface via the clathrin-mediated but not the caveolin-mediated pathway. When T or TGF-beta2 was added to Sertoli cell cultures with established functional BTB, both factors accelerated the kinetics of internalization of BTB proteins from the cell surface, perhaps above the migrating preleptotene spermatocyte, thereby opening the BTB. Likewise, T also enhanced the kinetics of recycling of internalized biotinylated proteins back to the cell surface, plausibly relocating these proteins beneath the migrating spermatocyte to reassemble the BTB. In contrast, TGF-beta2 targeted internalized biotinylated proteins to late endosomes for degradation, destabilizing the BTB. In summary, the transient opening of the BTB that facilitates germ cell movement is mediated via the differential effects of T and cytokines on the kinetics of endocytosis and recycling of integral membrane proteins at the BTB. The net result of these interactions, in turn, determines the steady-state protein levels at the Sertoli-Sertoli cell interface at the BTB. PMID:18192323

Yan, Helen H N; Mruk, Dolores D; Lee, Will M; Cheng, C Yan

2008-06-01

179

Effects of cyclic AMP and phorbol ester on transepithelial electrical resistance of Sertoli cell monolayers in two-compartment culture.  

PubMed

The effects of dibutyryl cyclic AMP [Bu)2cAMP) and phorbol ester (TPA), in the absence or presence of follicle-stimulating hormone (FSH) and/or testosterone, on the development of tight junctions by immature rat Sertoli cells (Sc) were investigated in vitro using the two-compartment culture system. The tight junction status was evaluated by repeated measurements of transepithelial electrical resistance (TER). Untreated cell monolayers developed stable TER of approximately 120 omega cm2 during 3 days of culture. Continuous presence of FSH (200 ng/ml) from day 1 onward significantly increased the TER up to approximately 300 omega cm2 after a transient (24-36 h) delay. The initial delay was prolonged to 3-4 days by the addition of 1-methyl-3-isobutylxanthine (MIX) (0.2 mM), whereas the subsequent increase of TER was significantly potentiated by the concomitant presence of testosterone (10 microM). Cholera toxin (CHT; 10 ng/ml) and forskolin (FR; 50 microM) mimicked these FSH effects. (Bu)2cAMP, at concentrations which maximally stimulated immunoactive inhibin secretion (100-500 microM), inhibited the initial TER increase and significantly decreased the TER level when added on days 1 and 5 of culture, respectively. In contrast, low concentrations of (Bu)2cAMP (4-20 microM) consistently stimulated the TER development, mimicking the stimulatory phase of FSH action. TPA (100 nM) alone had no effect on TER development, but potentiated the stimulatory effect of testosterone in a manner similar to FSH, CHT, FR or low concentrations of (Bu)2cAMP. These results demonstrate, for the first time, a concentration-dependent, dual effect of exogenous cAMP on the Sc function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1722179

Janecki, A; Jakubowiak, A; Steinberger, A

1991-11-01

180

Trafficking of sulfated glycoprotein-1 (prosaposin) to lysosomes or to the extracellular space in rat Sertoli cells.  

PubMed

Sulfated glycoprotein-1 (prosaposin) exists in 2 forms: a 65kDa form targeted to lysosomes and a 70kDa form secreted extracellularly. In order to understand the sorting and targeting mechanisms of the two forms of SGP-1, we have compared their maturation, processing, and secretion in rat Sertoli cells in vivo. Metabolic labeling experiments in vivo demonstrated that the 65kDa form is synthesized first, then post-translationally modified to the 70kDa form of SGP-1. Subcellular fractionation of testicular homogenate was used to obtain Golgi fractions containing up to 50-fold enrichment in galactosyltransferase. Permeabilization of enriched Golgi fractions with saponin released the 70kDa form, but did not affect the 65kDa protein. While excess free mannose 6-phosphate did not release lysosomal SGP-1, it released the 35kDa cathepsin L from Golgi membranes. Using quantitative electron-microscopic immunocytochemistry, the lysosomal contents of SGP-1 were shown to increase significantly after the administration of tunicamycin in vivo. Therefore, the trafficking of the 65kDa form of SGP-1 to the lysosomes appears to be independent of the M6P-receptor pathway. The 70kDa form of SGP-1 was found to aggregate within perforated Golgi fractions in a process which depends on low pH and calcium ions. We conclude that the targeting of the 65kDa form of SGP-1 to the lysosomes involves an early association with Golgi membrane that is independent of mannose 6-phosphate receptors. PMID:8593668

Igdoura, S A; Rasky, A; Morales, C R

1996-03-01

181

Differential interactions between transforming growth factor-beta3/TbetaR1, TAB1, and CD2AP disrupt blood-testis barrier and Sertoli-germ cell adhesion.  

PubMed

The biochemical basis that regulates the timely and selective opening of the blood-testis barrier (BTB) to migrating preleptotene/leptotene spermatocytes at stage VIII of the epithelial cycle in adult rat testes is virtually unknown. Recent studies have shown that cytokines (e.g. transforming growth factor (TGF)-beta3) may play a crucial role in this event. However, much of this information relies on the use of toxicants (e.g. CdCl(2)), making it difficult to relay these findings to normal testicular physiology. Here we report that overexpression of TGF-beta3 in primary Sertoli cells cultured in vitro indeed perturbed the tight junction (TJ) barrier with a concomitant decline in the production of BTB constituent proteins as follows: occludin, N-cadherin, and ZO-1. Additionally, local administration of TGF-beta3 to testes in vivo was shown to reversibly perturb the BTB integrity and Sertoli-germ cell adhesion via the p38 MAPK and ERK signaling pathways. Most importantly, the simultaneous activation of p38 and ERK signaling pathways is dependent on the association of the TGF-beta3-TbetaR1 complex with adaptors TAB1 and CD2AP because if TbetaR1 was associated preferentially with CD2AP, only Sertoli-germ cell adhesion was perturbed without compromising the BTB. Collectively, these data illustrate that local production of TGF-beta3, and perhaps other TGF-betas and cytokines, by Sertoli and germ cells into the microenvironment at the BTB during spermatogenesis transiently perturbs the BTB and Sertoli-germ cell adhesion to facilitate germ cell migration when the activated TbetaRI interacts with adaptors TAB1 and CD2AP. However, TGF-beta3 selectively disrupts Sertoli-germ cell adhesion in the seminiferous epithelium to facilitate germ cell migration without compromising BTB when TbetaRI interacts only with adaptor CD2AP. PMID:16617054

Xia, Weiliang; Mruk, Dolores D; Lee, Will M; Cheng, C Yan

2006-06-16

182

Isolation of murine lung endothelial cells  

PubMed Central

Several protocols for the isolation of endothelial cells (ECs) from murine lung have been described in the literature. We, however, encountered a number of problems while using these procedures that prevented us from consistently or reliably obtaining pure populations of ECs from the lungs of mice. By incorporating specific elements from previously published protocols, as well as adding some novel features, we developed a new strategy for isolating ECs from murine lung. In this approach, a suspension of lung cells is initially prepared from the lungs of 7- to 14-day-old mouse pups using procedures that prevent intravascular clotting and leukocyte activation, minimize mechanical trauma to the lung tissue, and limit exposure to the digesting enzymes. The resulting cell suspension is cultured for 2–3 days, trypsinized to produce a suspension of single cells, and then subjected to fluorescence-activated cell sorting using an anti-ICAM-2 antibody. The sorted cells are then plated and split 1:2 at each passage to maintain a high density of the cells. Using this approach, we have been able to isolate pure populations of ECs that were sustainable for extended periods in culture without the emergence of fibroblast overgrowth or the development of senescence. We believe the success of this approach will provide opportunities to take advantage of the large and growing number of knockout and transgenic mouse lines to investigate the endothelial-specific roles of targeted molecules in the pulmonary vasculature.

Fehrenbach, Melane L.; Cao, Gaoyuan; Williams, James T.; Finklestein, Jeffrey M.; DeLisser, Horace M.

2009-01-01

183

Pituitary follicle-stimulating hormone (FSH) induces CREM gene expression in Sertoli cells: involvement in long-term desensitization of the FSH receptor.  

PubMed Central

Transcription factor CREM (cAMP-responsive element modulator) plays a pivotal role in the nuclear response to cAMP in neuroendocrine cells. We have previously shown that follicle-stimulating hormone (FSH) directs CREM expression in male germ cells. The physiological importance of FSH in Sertoli cell function prompted us to analyze its effect on CREM expression in these cells. We observed a dramatic and specific increase in the CREM isoform ICER (inducible cAMP early repressor) expression, with a peak 4 h after FSH treatment of primary Sertoli cells. Interestingly, induced levels of ICER protein persist for a considerably longer time. Induction of the repressor ICER accompanies early down-regulation of the FSH receptor transcript, which leads to long-term desensitization. Here we show that ICER represses FSH receptor expression by binding to a CRE-like sequence in the regulatory region of the gene. Our results confirm the crucial role played by CREM in hormonal control and suggest its role in the long-term desensitization phenomenon of peptide membrane receptors. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6

Monaco, L; Foulkes, N S; Sassone-Corsi, P

1995-01-01

184

Retinol induces the ERK1/2-dependent phosphorylation of CREB through a pathway involving the generation of reactive oxygen species in cultured Sertoli cells.  

PubMed

The ability to regulate cell cycle progression and apoptosis through the activation of nuclear receptors and gene transcription has been generally accepted as a potential chemopreventive and therapeutic property of retinoids. However, recent studies suggest that retinol and related compounds can exert rapid and non-genomic effects, which may increase the production of reactive oxygen species (ROS) and lead to cell cycle disruption and malignant transformation. In this work, we report that, in Sertoli cells, retinol (7 microM) induces the Src-dependent activation of ERK1/2 MAPK and the ERK1/2-mediated phosphorylation of the transcription factor CREB. We found that these retinol-induced effects were completely blocked by the antioxidant Trolox 100 microM (a hydrophilic analogue of alpha-tocopherol), the hydroxyl radical scavenger mannitol (1 mM) and the addition of native superoxide dismutase (200 U/ml), and also that retinol increased the production of ROS and several other parameters indicative of oxidative stress during the same incubation periods in which ERK1/2 and CREB were phosphorylated. The activation of the ERK1/2-CREB pathway appears to be involved in the onset of some of the malignant effects caused by retinol in Sertoli cells since inhibition of ERK1/2 activation blocked the retinol-induced cell transformation and proliferation. PMID:16510265

Gelain, Daniel P; Cammarota, Martín; Zanotto-Filho, Alfeu; de Oliveira, Ramatis B; Dal-Pizzol, Felipe; Izquierdo, Iván; Bevilaqua, Lia R M; Moreira, José C F

2006-10-01

185

CNS Myelin and Sertoli Cell Tight Junction Strands Are Absent in Osp\\/Claudin-11 Null Mice  

Microsoft Academic Search

Oligodendrocyte-specific protein (OSP)\\/claudin-11 is a recently identified transmembrane protein found in CNS myelin and testis with unknown function. Herein we demonstrate that Osp null mice exhibit both neurological and reproductive deficits: CNS nerve conduction is slowed, hindlimb weakness is conspicuous, and males are sterile. Freeze fracture reveals that tight junction intramembranous strands are absent in CNS myelin and between Sertoli

Alexander Gow; Cherie M. Southwood; Jing Song Li; Milena Pariali; Gavin P. Riordan; Scott E. Brodie; John Danias; Jeff M. Bronstein; Bechara Kachar; Robert A. Lazzarini

1999-01-01

186

Isolation of murine adipose-derived stem cells.  

PubMed

Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Primary adipocytes grown in culture and derived from murine adipose tissue are essential for studying the mechanisms underlying adipocyte development and function. Herein, we describe methods for the isolation, expansion, and long-term storage of murine adipose-derived stem cells along with a protocol for inducing adipogenesis in this cell population. PMID:21082392

Yu, Gang; Wu, Xiying; Kilroy, Gail; Halvorsen, Yuan-Di C; Gimble, Jeffrey M; Floyd, Z Elizabeth

2011-01-01

187

Derivation of ES-like cell from neonatal mouse testis cells in autologous sertoli cells co-culture system  

PubMed Central

Background: Spermatogonial stem cell (SSC) is a self-renewing population of male adult stem cell. SSCs have a differentiation potential which are similar to embryonic stem cells. These Embryonic stem like (ES-like) cells can be a potential source for pluripotent cells for stem cell-based therapy. Objective: This study presents an economical and simple co-culture system for pluripotent stem cells generation from neonatal mouse testis Materials and Methods: Isolated testicular cells were cultured in DMEM/F12. Characteristics of the isolated cells and obtained ES-like cell were immune-cytochemically confirmed by examining the presence of PLZF, vimentin, Oct4 and Nanog protein. Expression of the pluripotency and germ-cell specific genes was analyzed by qPCR in derived ES-like colony and SSCs respectively. Results: The experiment results indicated that our method of obtaining pluripotent ES-like cells from spermatogonial cells (SCs) is simpler than the described methods. ES-like cells were immunopositive for pluripotency markers. ES-like cell qPCR results indicated signi?cant increase in pluripotency genes expression and signi?cant decrease in germ cell-speci?c genes expression. Conclusion: The results indicated that ES-like cell with pluripotency characteristic were generated from freshly isolated spermatogonial cells. The pluripotent stem cells provide a cellular reservoir usable for regenerative medicine instead of embryonic stem cells. This article extracted from Ph.D. thesis. (Setareh Javanmardi)

Asadi, Mohammad Hossein; Javanmardi, Setareh; Movahedin, Mansoureh

2014-01-01

188

Maternal undernutrition does not alter Sertoli cell numbers or the expression of key developmental markers in the mid-gestation ovine fetal testis  

PubMed Central

Background The aim of this study was to determine the effects of maternal undernutrition on ovine fetal testis morphology and expression of relevant histological indicators. Maternal undernutrition, in sheep, has been reported, previously, to alter fetal ovary development, as indicated by delayed folliculogenesis and the altered expression of ovarian apoptosis-regulating gene products, at day 110 of gestation. It is not known whether or not maternal undernutrition alters the same gene products in the day 110 fetal testis. Design and methods Mature Scottish Blackface ewes were fed either 100% (Control; C) or 50% (low; L) of estimated metabolisable energy requirements of a pregnant ewe, from mating to day 110 of gestation. All pregnant ewes were euthanized at day 110 and a sub-set of male fetuses was randomly selected (6 C and 9?L) for histology studies designed to address the effect of nutritional state on several indices of testis development. Sertoli cell numbers were measured using a stereological method and Ki67 (cell proliferation index), Bax (pro-apoptosis), Mcl-1 (anti-apoptosis), SCF and c-kit ligand (development and apoptosis) gene expression was measured in Bouins-fixed fetal testis using immunohistochemistry. Results No significant differences were observed in numbers of Sertoli cells or testicular Ki67 positive cells. The latter were localised to the testicular cords and interstitium. Bax and Mcl-1 were localised specifically to the germ cells whereas c-kit was localised to both the cords and interstitium. SCF staining was very sparse. No treatment effects were observed for any of the markers examined. Conclusions These data suggest that, unlike in the fetal ovary, maternal undernutrition for the first 110?days of gestation affects neither the morphology of the fetal testis nor the expression of gene products which regulate apoptosis. It is postulated that the effects of fetal undernutrition on testis function may be expressed through hypothalamic-pituitary changes.

2013-01-01

189

Tumor necrosis factor {alpha} reversibly disrupts the blood-testis barrier and impairs Sertoli-germ cell adhesion in the seminiferous epithelium of adult rat testes.  

PubMed

The timely restructuring of the blood-testis barrier (BTB) that facilitates the migration of preleptotene and leptotene spermatocytes from the basal to the adluminal compartment in the seminiferous epithelium of adult rat testes, which occurs at late stage VII through early stage VIII of the epithelial cycle, is a crucial cellular event of spermatogenesis. However, the regulation of BTB dynamics at the biochemical level remains elusive. In this study, tumor necrosis factor alpha (TNFalpha), a secretory product of Sertoli and germ cells in rat testes, was shown to affect junction dynamics in vivo. Following an acute administration of recombinant TNFalpha directly to adult rat testes in vivo at 0.5 and 2 mug/testis (with a body weight ~300 g), this treatment significantly and transiently disrupted the BTB. It also transiently inhibited the steady-state protein levels of occludin, zonula occludens-1, and N-cadherin, but not junction adhesion molecule-A, alpha-, and beta-catenin in testes at the BTB site as illustrated by immunoblottings, immunohistochemistry, electron microscopy, and fluorescent microscopy. This transient disruption of the BTB integrity induced by TNFalpha treatment was further demonstrated by a functional test to assess the passage of a fluorescent dye (e.g. fluorescein-5-isothiocyanate) from the systemic circulation to the adluminal compartment. Additionally, both the phosphorylated-Ser/Thr protein kinase activated by MAP kinase kinase (p-p38) and phosphorylated-externally regulated kinase (p-ERK) mitogen -activated protein kinase-signaling pathways were transiently activated. Collectively, these data coupled with the recently published in vitro studies have illustrated that the BTB is likely utilizing a novel mechanism in which localized production of TNFalpha by Sertoli and germ cells into the microenvironment at the basal compartment facilitates the timely restructuring ('opening'?) of the BTB during spermatogenesis to facilitate germ cell migration. PMID:16899565

Li, Michelle W M; Xia, Weiliang; Mruk, Dolores D; Wang, Claire Q F; Yan, Helen H N; Siu, Michelle K Y; Lui, Wing-Yee; Lee, Will M; Cheng, C Yan

2006-08-01

190

Nuclear Nonhistone Proteins in Murine Melanoma Cells  

PubMed Central

Nuclear nonhistone proteins (NHP's) have been implicated as regulatory agents involved in controlling genetic expression. Utilizing murine melanoma cells, we describe a method for isolating and fractionating NHP's which greatly increases the yield of these proteins as well as the level of resolution required for detecting small differences in particular NHP's. Mouse melanoma cells were grown in medium labeled with [3H]leucine. Following 48 hr of incubation, the cells were harvested and nuclei isolated. The NHP's were extracted from the nuclei in a series of steps which yielded four major fractions: NHP1, NHP2, NHP3, NHP4. This method solubilized 80-90% of the protein from the nuclear homogenate. The NHP fractions were then separated on DEAE-cellulose columns in a series of salt steps increasing in concentration from 0.05 to 0.50 M NaCl, followed by steps of 2 M NaCl and 4 and 7 M guanidine-hydrochloride. The 40 NHP fractions eluted from these columns were further separated on polyacrylamide-SDS gels and ranged in molecular weight from 9000 to 110,000 daltons. Differences were observed in the electrophoretic pattern of each of these 40 fractions. The high resolution of these fractionation procedures greatly enhances the possibility of observing small changes in proteins which may play a role in gene regulation. ImagesFIG. 2FIG. 5

Wikswo, Muriel A.; Mcguire, Joseph S.; Shansky, Janet E.; Boshes, Roger A.

1976-01-01

191

Murine Mueller cells are progenitor cells for neuronal cells and fibrous tissue cells  

SciTech Connect

Mammalian Mueller cells have been reported to possess retinal progenitor cell properties and generate new neurons after injury. This study investigates murine Mueller cells under in vitro conditions for their capability of dedifferentiation into retinal progenitor cells. Mueller cells were isolated from mouse retina, and proliferating cells were expanded in serum-containing medium. For dedifferentiation, the cultured cells were transferred to serum-replacement medium (SRM) at different points in time after their isolation. Interestingly, early cell passages produced fibrous tissue in which extracellular matrix proteins and connective tissue markers were differentially expressed. In contrast, aged Mueller cell cultures formed neurospheres in SRM that are characteristic for neuronal progenitor cells. These neurospheres differentiated into neuron-like cells after cultivation on laminin/ornithine cell culture substrate. Here, we report for the first time that murine Mueller cells can be progenitors for both, fibrous tissue cells and neuronal cells, depending on the age of the cell culture.

Florian, Christian; Langmann, Thomas; Weber, Bernhard H.F. [Institute of Human Genetics, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg (Germany); Morsczeck, Christian [Institute of Human Genetics, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg (Germany)], E-mail: Christian.morsczeck@klinik.uni-regensburg.de

2008-09-19

192

Telomere sister chromatid exchange in telomerase deficient murine cells  

SciTech Connect

We have recently demonstrated that several types of genomic rearrangements (i.e., telomere sister chromatid exchange (T-SCE), genomic-SCE, or end-to-end fusions) were more often detected in long-term cultured murine telomerase deficient embryonic stem (ES) cells than in freshly prepared murine splenocytes, even through they possessed similar frequencies of critically short telomeres. The high rate of genomic rearrangements in telomerase deficient ES cells, when compared to murine splenocytes, may reflect the cultured cells' gained ability to protect chromosome ends with eroded telomeres allowing them to escape 'end crisis'. However, the possibility that ES cells were more permissive to genomic rearrangements than other cell types or that differences in the microenvironment or genetic background of the animals might consequentially determine the rate of T-SCEs or other genomic rearrangements at critically short telomeres could not be ruled out.

Wang, Yisong [ORNL; Giannone, Richard J [ORNL; Liu, Yie [ORNL

2005-01-01

193

Activation of the Murine Sarcoma Virus Genome After Infection with the Murine Leukemia Virus as Determined by Cell Agglutination  

PubMed Central

Non-virus-producing NIH/3T3 cells transformed by the murine sarcoma virus are agglutinated by conconavalin A to the same low level as normal NIH/3T3 cells. Infection with the murine leukemia virus greatly increases the agglutination of transformed cells but not that of normal cells. These data suggest that the morphological expression of cell transformation and the surface alterations associated with increased cell agglutination are controlled by the expressions of different sarcoma virus genes.

Salzberg, Samuel; Green, Maurice

1974-01-01

194

Embryonic Stem Cell Virus, a Recombinant Murine Retrovirus with Expression in Embryonic Stem Cells  

Microsoft Academic Search

The expression of Moloney murine leukemia virus and vectors derived from it is restricted in undifferentiated mouse embryonal carcinoma and embryonal stem (ES) cells. We have developed a retroviral vector, the murine embryonic stem cell virus (MESV), that is active in embryonal carcinoma and ES cells. MESV was derived from a retroviral mutant [PCC4-cell-passaged myeloproliferative sarcoma virus (PCMV)] expressed in

Manuel Grez; Ercan Akgun; Frank Hilberg; Wolfram Ostertag

1990-01-01

195

Regulation of follitropin-sensitive adenylate cyclase by stimulatory and inhibitory forms of the guanine nucleotide regulatory protein in immature rat Sertoli cells  

SciTech Connect

Studies have been designed to examine the role of guanine nucleotides in mediating FSH-sensitive adenylate cyclase activity in Sertoli cell plasma membranes. Analysis of ({sup 3}H)GDP binding to plasma membranes suggested a single high affinity site with a K{sub d} = 0.24 uM. Competition studies indicated that GTP{sub {gamma}}S was 7-fold more potent than GDP{sub {beta}}S. Bound GDP could be released by FSH in the presence of GTP{sub {gamma}}S, but not by FSH alone. Adenylate cyclase activity was enhanced 5-fold by FSH in the presence of GTP. Addition of GDP{sub {beta}}S to the activated enzyme (FSH plus GTP) resulted in a time-dependent decay to basal activity within 20 sec. GDP{sub {beta}}S competitively inhibited GTP{sub {gamma}}S-stimulated adenylate cyclase activity with a K{sub i} = 0.18 uM. Adenylate cyclase activity was also demonstrated to be sensitive to the nucleotide bound state. In the presence of FSH, only the GTP{sub {gamma}}S-bound form persisted even if GDP{sub {beta}}S previously occupied all available binding sites. Two membrane proteins, M{sub r} = 43,000 and 48,000, were ADP{centered dot}ribosylated using cholera toxin and labeling was enhanced 2 to 4-fold by GTP{sub {gamma}}S but not by GDP{sub {beta}}S. The M{sub r} = 43,000 and 48,000 proteins represented variant forms of G{sub S}. A single protein of M{sub r} = 40,000 (G{sub i}) was ADP-ribosylated by pertussis toxin in vitro. GTP inhibited forskolin-stimulated adenylate cyclase activity with an IC{sub 50} = 0.1 uM. The adenosine analog, N{sup 6}{centered dot}phenylisopropyl adenosine enhanced GTP inhibition of forskolin-stimulated adenylate cyclase activity by an additional 15%. GTP-dependent inhibition of forskolin-sensitive adenylate cyclase activity was abolished in membranes prepared from Sertoli cells treated in culture with pertussis toxin.

Johnson, G.P.

1987-01-01

196

Elderly Men Have Low Levels of Anti-M?llerian Hormone and Inhibin B, but with High Interpersonal Variation: A Cross-Sectional Study of the Sertoli Cell Hormones in 615 Community-Dwelling Men  

PubMed Central

The Sertoli cells of the testes secrete anti-Müllerian hormone (Müllerian inhibiting Substance, AMH) and inhibin B (InhB). AMH triggers the degeneration of the uterine precursor in male embryos, whereas InhB is part of the gonadal-pituitary axis for the regulation of sperm production in adults. However, both hormones are also putative regulators of homeostasis, and age-related changes in these hormones may therefore be important to the health status of elderly men. The levels of AMH in elderly men are unknown, with limited information being available about age-related changes in InhB. We have therefore used ELISAs to measure Sertoli cell hormone levels in 3 cohorts of community-dwelling men in New Zealand. In total, 615 men were examined, 493 of which were aged 65 or older. Serum AMH and InhB levels inversely correlated with age in men older than 50 years (p<0.001) but not in the younger men. A minority of elderly men had undetectable levels of AMH and InhB. The variation in hormone levels between similarly aged men increased with the age of men. AMH and InhB partially correlated with each other as expected (r?=?0.48, p<0.001). However, the ratio of the two Sertoli hormones varied significantly between men, with this variation increasing with age. Elderly men selected for the absence of cardiovascular disease had AMH levels similar to those of young men whereas their InhB levels did not differ from aged-matched controls. These data suggests that Sertoli cell number and function changes with age, but with the extent and nature of the changes varying between men.

Chong, Yih Harng; Dennis, Nicola A.; Connolly, Martin J.; Teh, Ruth; Jones, Gregory T.; van Rij, Andre M.; Farrand, Stephanie; Campbell, A. John; MLennan, Ian S.

2013-01-01

197

Improving in vitro Sertoli cell/Gonocyte co-culture model for assessing male reproductive toxicity: lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates  

PubMed Central

Gonocytes exist in the neonatal testis and represent a transient population of male germ-line stem cells. It has been shown that stem cell self-renewal and progeny production is probably controlled by the neighboring differentiated cells and extracellular matrix (ECM) in vivo known as niches. Recently, we developed an in vitro three-dimensional (3D) Sertoli cell/Gonocyte coculture (SGC) model with ECM overlay, which creates an in vivo-like niche and supports germ-line stem cell functioning within a 3D environment. In this study, we applied morphological and cytotoxicity evaluations, as well as microarray-based gene expression to examine the effects of different phthalate esters (PE) on this model. Known in vivo male developmentally toxic PEs (DTPE) and developmentally nontoxic PEs (DNTPE) were evaluated. We observed that DTPE induced significantly greater dose-dependent morphological changes, a decrease in cell viability and an increase in cytotoxicity compared to those treated with DNTPE. Moreover, gene expression was more greatly altered by DTPE than by DNTPE and non-supervised cluster analysis allowed the discrimination of DTPE from the DNTPE. Our systems-based GO-Quant analysis showed significant alterations in gene pathways involved in cell cycle, phosphate transport and apoptosis regulation with DTPE but not with DNTPE treatment. Disruptions of steroidogenesis related-gene expression such as Star, Cyp19a1, Hsd17b8, and Nr4a3 were observed in the DTPE group, but not in the DNTPE group. In summary, our observation on cell viability, cytotoxicity, and microarray-based gene expression analysis induced by PEs demonstrate that our in vitro 3D-SGC system mimicked in vivo responses for PEs and suggests that the 3D-SGC system might be useful in identifying developmental reproductive toxicants.

Yu, Xiaozhong; Hong, SungWoo; Moreira, Estefania G; Faustman, Elaine M

2009-01-01

198

Improving in vitro Sertoli cell/gonocyte co-culture model for assessing male reproductive toxicity: Lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates  

SciTech Connect

Gonocytes exist in the neonatal testis and represent a transient population of male germ-line stem cells. It has been shown that stem cell self-renewal and progeny production is probably controlled by the neighboring differentiated cells and extracellular matrix (ECM) in vivo known as niches. Recently, we developed an in vitro three-dimensional (3D) Sertoli cell/gonocyte co-culture (SGC) model with ECM overlay, which creates an in vivo-like niche and supports germ-line stem cell functioning within a 3D environment. In this study, we applied morphological and cytotoxicity evaluations, as well as microarray-based gene expression to examine the effects of different phthalate esters (PE) on this model. Known in vivo male developmentally toxic PEs (DTPE) and developmentally non-toxic PEs (DNTPE) were evaluated. We observed that DTPE induced significantly greater dose-dependent morphological changes, a decrease in cell viability and an increase in cytotoxicity compared to those treated with DNTPE. Moreover, the gene expression was more greatly altered by DTPE than by DNTPE and non-supervised cluster analysis allowed the discrimination of DTPE from the DNTPE. Our systems-based GO-Quant analysis showed significant alterations in the gene pathways involved in cell cycle, phosphate transport and apoptosis regulation with DTPE but not with DNTPE treatment. Disruptions of steroidogenesis related-gene expression such as Star, Cyp19a1, Hsd17b8, and Nr4a3 were observed in the DTPE group, but not in the DNTPE group. In summary, our observation on cell viability, cytotoxicity, and microarray-based gene expression analysis induced by PEs demonstrate that our in vitro 3D-SGC system mimicked in vivo responses for PEs and suggests that the 3D-SGC system might be useful in identifying developmental reproductive toxicants.

Yu Xiaozhong; Hong, Sung Woo [Institute for Risk Analysis and Risk Communication, Department of Environmental and Occupational Health Sciences, University of Washington, 4225 Roosevelt Way NE, Suite 100, Seattle, WA 98105-6099 (United States); Moreira, Estefania G. [Institute for Risk Analysis and Risk Communication, Department of Environmental and Occupational Health Sciences, University of Washington, 4225 Roosevelt Way NE, Suite 100, Seattle, WA 98105-6099 (United States); Dept of Physiological Sciences, State University of Londrina (UEL), Londrina, PR (Brazil); Faustman, Elaine M. [Institute for Risk Analysis and Risk Communication, Department of Environmental and Occupational Health Sciences, University of Washington, 4225 Roosevelt Way NE, Suite 100, Seattle, WA 98105-6099 (United States)], E-mail: faustman@u.washington.edu

2009-09-15

199

?-Benzene hexachloride induces apoptosis of rat Sertoli cells through generation of reactive oxygen species and activation of JNKs and FasL.  

PubMed

?-benzene hexachloride (?-BHC), the major metabolite of benzene-hexachloride (BHC), is a weak estrogen-like chemical. It is a known persistent organic pollutant and male reproductive toxicant. However, the mechanism by which ?-BHC exposure causes male reproductive toxicity remains unknown. In the present study, rat Sertoli cells were used to investigate the molecular mechanism involved in ?-BHC-induced toxicity in male reproductive system. The results indicated that ?-BHC exposure at over 30 ?M showed the induction of apoptotic cell death. ?-BHC could induce elevation in reactive oxygen species (ROS) generation, increase in the leakage rate of LDH and MDA level, and decrease in SOD activity. In addition, there was an increase in the cellular levels of phospho-JNKs and FasL in the ?-BHC-induced apoptosis; and a significant reduction of procaspase-3 and -8 was observed over 30-?M ?-BHC treatment. The translocation of NF-?B enhanced with the increase of concentration of ?-BHC. Furthermore, NAC administration, a scavenger of ROS, reversed ?-BHC-induced apoptosis effects via inhibition of JNKs activation, FasL expression, and NF-?B translocation. These results lead us to speculate that ROS generation may play a critical role in the initiation of ?-BHC-induced apoptosis by activation of the JNKs, translocation of NF-?B, expression of FasL, and further activation of caspase cascade. PMID:19760616

Shi, Yuqin; Song, Yang; Wang, Yuping; Wang, Yinan; Liang, Xianmin; Hu, Yafei; Yu, Haige; Guan, Xia; Cheng, Jin; Yang, Kedi

2011-04-01

200

Catecholamines in murine bone marrow derived mast cells  

Microsoft Academic Search

Cultured murine bone marrow derived mast cells (BMMC) were found to store high levels of dopamine (3753±844 pg\\/107 cells) and occasionally produce norepinephrine and epinephrine. The catecholamine synthesis inhibitor, ?-methyl-para-tyrosine, decreased intracellular catecholamine concentrations, and activation with ionomycin stimulated dopamine release. Neither dopaminergic receptor antagonists nor exogenous dopamine ?10 ?M affected IL-3-induced cell proliferation. High exogenous dopamine (20–100 ?M) decreased

Jessica G Freeman; John J Ryan; Christopher P Shelburne; Daniel P Bailey; L. Andrew Bouton; Nedathur Narasimhachari; Jos Domen; Nathalie Siméon; François Couderc; Jennifer K Stewart

2001-01-01

201

SYNCHRONIZATION OF RAPID GLOBIN EXPRESSION IN MURINE ERYTHROLEUKEMIC CELLS  

EPA Science Inventory

The addition of butyric acid (BA) to murine erythroleukemia cells (MELC) produces the expression of primarily A and E2 hemoglobins while DMSO incubation produces the expression of primarily A hemoglobin. Preincubation of MELC with DMSO followed by BA induction accelerates the exp...

202

[Cardiomyocyte differentiation of individual clones murine induced pluripotent stem cells].  

PubMed

Cardiomyocyte differentiation of certain clones of murine induced pluripotent stem cells (iPS) was estimated. iPS were obtained due to reprogramming of murine embryonic fibroblasts with transposon-based Sleeping beauty plasmids as sene delivery systems. Differentiation was performed in suspension culture and in attached to tissue-culture plates embryoid bodies (EBs). Ascorbic acid was applied as inductor. According to the obtained results, the differentiation was tenfold more effective in attached EBs. Ascorbic acid stimulated the generation of cardiomiocytes. PMID:23957159

Malysheva, S V; Budash, H V; Bil'ko, N M; Heschheller, J

2013-01-01

203

T Cell Integrin Overexpression as a Model of Murine Autoimmunity  

PubMed Central

Integrin adhesion molecules have important adhesion and signaling functions. They also play a central role in the pathogenesis of many autoimmune diseases. Over the past few years we have described a T cell adoptive transfer model to investigate the role of T cell integrin adhesion molecules in the development of autoimmunity. This report summarizes the methods we used in establishing this murine model. By treating murine CD4+ T cells with DNA hypomethylating agents and by transfection we were able to test the in vitro effects of integrin overexpression on T cell autoreactive proliferation, cytotoxicity, adhesion and trafficking. Furthermore, we showed that the ability to induce in vivo autoimmunity may be unique to the integrin lymphocyte function associated antigen-1 (LFA-1).

Ray, Donna; Mo, Ru Ran; Chen, Jun

2003-01-01

204

Expression of ia antigens by murine keratinizing epithelial langerhans cells  

Microsoft Academic Search

Immunofluorescent and immunoelectron-microscopic staining methods were utilized to investigate the localization of Ia antigens\\u000a in murine keratinizing epithelia. Approximately 3–5% of epidermal cells were shown to be Ia positive. Only dendritic Langerhans\\u000a cells in the interfollicular epidermis and outer root sheaths were found to express Ia antigens. These Ia determinants were\\u000a shown to be controlled by both theI- A andI-

Geoffrey Rowden; Terence M. Phillips; Terry L. Delovitch

1978-01-01

205

Human pontine glioma cells can induce murine tumors.  

PubMed

Diffuse intrinsic pontine glioma (DIPG), with a median survival of only 9 months, is the leading cause of pediatric brain cancer mortality. Dearth of tumor tissue for research has limited progress in this disease until recently. New experimental models for DIPG research are now emerging. To develop preclinical models of DIPG, two different methods were adopted: cells obtained at autopsy (1) were directly xenografted orthotopically into the pons of immunodeficient mice without an intervening cell culture step or (2) were first cultured in vitro and, upon successful expansion, injected in vivo. Both strategies resulted in pontine tumors histopathologically similar to the original human DIPG tumors. However, following the direct transplantation method all tumors proved to be composed of murine and not of human cells. This is in contrast to the indirect method that included initial in vitro culture and resulted in xenografts comprising human cells. Of note, direct injection of cells obtained postmortem from the pons and frontal lobe of human brains not affected by cancer did not give rise to neoplasms. The murine pontine tumors exhibited an immunophenotype similar to human DIPG, but were also positive for microglia/macrophage markers, such as CD45, CD68 and CD11b. Serial orthotopic injection of these murine cells results in lethal tumors in recipient mice. Direct injection of human DIPG cells in vivo can give rise to malignant murine tumors. This represents an important caveat for xenotransplantation models of DIPG. In contrast, an initial in vitro culture step can allow establishment of human orthotopic xenografts. The mechanism underlying this phenomenon observed with direct xenotransplantation remains an open question. PMID:24777482

Caretti, Viola; Sewing, A Charlotte P; Lagerweij, Tonny; Schellen, Pepijn; Bugiani, Marianna; Jansen, Marc H A; van Vuurden, Dannis G; Navis, Anna C; Horsman, Ilona; Vandertop, W Peter; Noske, David P; Wesseling, Pieter; Kaspers, Gertjan J L; Nazarian, Javad; Vogel, Hannes; Hulleman, Esther; Monje, Michelle; Wurdinger, Thomas

2014-06-01

206

MURINE MODELS OF LUPUS INDUCED BY HYPOMETHYLATED T CELLS  

PubMed Central

CD4+ T cell DNA hypomethylation may contribute to the development of drug induced and idiopathic human lupus. Inhibiting DNA methylation in mature CD4+ T cells causes MHC-specific autoreactivity in vitro. The lupus-inducing drugs hydralazine and procainamide also inhibit T cell DNA methylation and induce autoreactivity, and T cells from patients with active lupus have hypomethylated DNA and a similarly autoreactive T cell subset. Further, T cells treated with DNA methylation inhibitors demethylate the same sequences that demethylate in T cells from patients with active lupus. The pathologic significance of the autoreactivity induced by inhibiting T cell DNA methylation has been tested by treating murine T cells in vitro with drugs which modify DNA methylation, then injecting the cells into syngeneic female mice. Mice receiving CD4+ T cells demethylated by a variety of agents including procainamide and hydralazine develop a lupus-like disease. Further, transgenic mice with an inducible T cell DNA methylation defect also develop lupus-like autoimmunity. This chapter describes the protocols for inducing autoreactivity in murine T cells in vitro, and inducing autoimmunity in vivo using an adoptive transfer approach or transgenic animal models.

Richardson, Bruce; Sawalha, Amr H; Ray, Donna; Yung, Raymond

2014-01-01

207

Follicle-stimulating hormone receptor-mediated uptake of sup 45 Ca sup 2+ by cultured rat Sertoli cells does not require activation of cholera toxin- or pertussis toxin-sensitive guanine nucleotide binding proteins or adenylate cyclase  

SciTech Connect

We have previously reported that FSH stimulates flux of 45Ca2+ into cultured Sertoli cells from immature rats via voltage-sensitive and voltage-independent calcium channels. In the present study, we show that this effect of FSH does not require cholera toxin (CT)- or pertussis toxin (PT)-sensitive guanine nucleotide binding (G) protein or activation of adenylate cyclase (AC). Significant stimulation of 45Ca2+ influx was observed within 1 min, and maximal response (3.2-fold over basal levels) was achieved within 2 min after exposure to FSH. FSH-stimulated elevations in cellular cAMP paralleled increases in 45Ca2+ uptake, suggesting a possible coupling of AC activation to 45Ca2+ influx. (Bu)2cAMP, however, was not able to enhance 45Ca2+ uptake over basal levels at a final concentration of 1000 microM, although a concentration-related increase in androstenedione conversion to estradiol was evident. Exposure of Sertoli cells to CT (10 ng/ml) consistently stimulated basal levels of androstenedione conversion to estradiol but had no effect on basal levels of 45Ca2+ uptake. Similarly, CT had no effect on FSH-induced 45Ca2+ uptake, but potentiated FSH-stimulated estradiol synthesis. PT (10 ng/ml) augmented basal and FSH-stimulated estradiol secretion without affecting 45Ca2+ influx. The adenosine analog N6-phenylisopropyladenosine, which binds to Gi-coupled adenosine receptors on Sertoli cells, inhibited FSH-stimulated androgen conversion to estradiol in a dose-related (1-1000 nM) manner, but FSH-stimulated 45Ca2+ influx remained unchanged. Our results show that in contrast to FSH-stimulated estradiol synthesis, the flux of 45Ca2+ into Sertoli cells in response to FSH is not mediated either directly or indirectly by CT- or PT-sensitive G protein, nor does it require activation of AC. Our data further suggest that the FSH receptor itself may function as a calcium channel.

Grasso, P.; Reichert, L.E. Jr. (Albany Medical College, NY (USA))

1990-08-01

208

Murine Cell Glycolipids Customization by Modular Expression of Glycosyltransferases  

PubMed Central

Functional analysis of glycolipids has been hampered by their complex nature and combinatorial expression in cells and tissues. We report an efficient and easy method to generate cells with specific glycolipids. In our proof of principle experiments we have demonstrated the customized expression of two relevant glycosphingolipids on murine fibroblasts, stage-specific embryonic antigen 3 (SSEA-3), a marker for stem cells, and Forssman glycolipid, a xenoantigen. Sets of genes encoding glycosyltansferases were transduced by viral infection followed by multi-color cell sorting based on coupled expression of fluorescent proteins.

Cid, Emili; Yamamoto, Miyako; Buschbeck, Marcus; Yamamoto, Fumiichiro

2013-01-01

209

FUNCTIONAL HETEROGENEITY OF MURINE LYMPHOID CELLS  

PubMed Central

An appreciable percent (3–14%) of the lymphocyte-like cells of the mouse spleen lack both the ?-isoantigen and sufficient surface immunoglobulin to be detected by conventional immunofluorescence or autoradiographic procedures. These ?-,Ig- cells are increased in frequency after treatment of mice with antithymocyte serum or in mice that have been thymectomized, irradiated (850 R), and reconstituted with bone marrow cells. Moreover, in chimeric C57BL/6 mice in which the T cells are derived from (BALB/c x C57BL/6)F1 donors, ?-,Ig- cells also lack BALB/c histocompatibility antigens. These experiments indicate that ?-,Ig- cells are not ?- T lymphocytes. Removal of complement receptor lymphocytes from spleen cell populations increases the frequency of ?-,Ig- cells, indicating that such cells lack the complement receptor. Partially purified populations of ?-,Ig- cells have been obtained by cytolysis by anti-?- and anti-?-antibody and complement and by density gradient ultracentrifugation. These cells closely resemble lymphocytes in morphology. The only exceptional feature is the existence of prominent nucleoli. The ?-,Ig- cells lack hemoglobin and endogenous peroxidases, are not actively phagocytic, and do not adhere to glass. This suggests they are not of the erythroid, myeloid, or monocytoid lines. [3H]Thymidine labeling studies indicate that ?-,Ig- cells are members of a relatively slowly dividing cell pool. Whether ?-,Ig- cells are members of the "classical" B lymphocyte line or belong to another, as yet undescribed, lineage is not yet certain.

Stobo, John D.; Rosenthal, Alan S.; Paul, William E.

1973-01-01

210

Murine Oligodendroglial Cells Express Nerve Growth Factor  

Microsoft Academic Search

The studies reported here present evidence for the expression of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) by an oligodendroglial cell line and of NGF by oligodendrocytes in mouse primary culture. An immortalized oligodendroglial cell line (N19) expressing markers for immature oligodendrocytes stimulated PC12 cells to elaborate processes. Polymerase chain reaction analysis with degenerate primers indicated that the

Sujatha Byravan; Lyndon M. Foster; Tommy Phan; A. Neil Verity; Anthony T. Campagnoni

1994-01-01

211

Antagonistic Effects of a Mixture of Low-Dose Nonylphenol and Di-N-Butyl Phthalate (Monobutyl Phthalate) on the Sertoli Cells and Serum Reproductive Hormones in Prepubertal Male Rats In Vitro and In Vivo  

PubMed Central

The estrogenic chemical nonylphenol (NP) and the antiandrogenic agent di-n-butyl phthalate (DBP) are regarded as widespread environmental endocrine disruptors (EDCs) which at high doses in some species of laboratory animals, such as mice and rats, have adverse effects on male reproduction and development. Given the ubiquitous coexistence of various classes of EDCs in the environment, their combined effects warrant clarification. In this study, we attempted to determine the mixture effects of NP and DBP on the testicular Sertoli cells and reproductive endocrine hormones in serum in male rats based on quantitative data analysis by a mathematical model. In the in vitro experiment, monobutyl phthalate (MBP), the active metabolite of DBP, was used instead of DBP. Sertoli cells were isolated from 9-day-old Sprague-Dawley rats followed by treatment with NP and MBP, singly or combined. Cell viability, apoptosis, necrosis, membrane integrity and inhibin-B concentration were tested. In the in vivo experiment, rats were gavaged on postnatal days 23–35 with a single or combined NP and DBP treatment. Serum reproductive hormone levels were recorded. Next, Bliss Independence model was employed to analyze the quantitative data obtained from the in vitro and in vivo investigation. Antagonism was identified as the mixture effects of NP and DBP (MBP). In this study, we demonstrate the potential of Bliss Independence model for the prediction of interactions between estrogenic and antiandrogenic agents.

Xiang, Zou; Qian, Weiping; Han, Xiaodong; Li, Dongmei

2014-01-01

212

Extrathymic development of murine T cells after bone marrow transplantation  

PubMed Central

Restoring T cell competence is a significant clinical challenge in patients whose thymic function is severely compromised due to age or cytoreductive conditioning. Here, we demonstrate in mice that mesenteric LNs (MLNs) support extrathymic T cell development in euthymic and athymic recipients of bone marrow transplantation (BMT). Furthermore, in aged murine BMT recipients, the contribution of the MLNs to the generation of T cells was maintained, while the contribution of the thymus was significantly impaired. Thymic impairment resulted in a proportional increase in extrathymic-derived T cell progenitors. Extrathymic development in athymic recipients generated conventional naive TCR?? T cells with a broad V? repertoire and intact functional and proliferative potential. Moreover, in the absence of a functional thymus, immunity against known pathogens could be augmented using engineered precursor T cells with viral specificity. These findings demonstrate the potential of extrathymic T cell development for T cell reconstitution in patients with limited thymic function.

Holland, Amanda M.; Zakrzewski, Johannes L.; Tsai, Jennifer J.; Hanash, Alan M.; Dudakov, Jarrod A.; Smith, Odette M.; West, Mallory L.; Singer, Natalie V.; Brill, Jessie; Sun, Joseph C.; van den Brink, Marcel R.M.

2012-01-01

213

Nanoelectroablation therapy for murine basal cell carcinoma  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer Nanoelectroablation is a new, non-thermal therapy that triggers apoptosis in tumors. Black-Right-Pointing-Pointer Low energy, ultrashort, high voltage pulses ablate the tumor with little or no scar. Black-Right-Pointing-Pointer Nanoelectroablation eliminates 99.8% of the BCC but may leave a few remnants behind. Black-Right-Pointing-Pointer Pilot clinical trials on human BCCs are ongoing and leave no remnants in most cases. -- Abstract: When skin tumors are exposed to non-thermal, low energy, nanosecond pulsed electric fields (nsPEF), apoptosis is initiated both in vitro and in vivo. This nanoelectroablation therapy has already been proven effective in treating subdermal murine allograft tumors. We wanted to determine if this therapy would be equally effective in the treatment of autochthonous BCC tumors in Ptch1{sup +/-}K14-Cre-ER p53 fl/fl mice. These tumors are similar to human BCCs in histology and in response to drug therapy . We have treated 27 BCCs across 8 mice with either 300 pulses of 300 ns duration or 2700 pulses of 100 ns duration, all at 30 kV/cm and 5-7 pulses per second. Every nsPEF-treated BCC began to shrink within a day after treatment and their initial mean volume of 36 {+-} 5 (SEM) mm{sup 3} shrunk by 76 {+-} 3% over the ensuing two weeks. After four weeks, they were 99.8% ablated if the size of the treatment electrode matched the tumor size. If the tumor was larger than the 4 mm wide electrode, multiple treatments were needed for complete ablation. Treated tumors were harvested for histological analysis at various times after treatment and exhibited apoptosis markers. Specifically, pyknosis of nuclei was evident as soon as 2 days after nsPEF treatment, and DNA fragmentation as detected via TUNEL staining was also evident post treatment. Nanoelectroablation is effective in triggering apoptosis and remission of radiation-induced BCCs with a single 6 min-long treatment of 2700 pulses.

Nuccitelli, Richard, E-mail: rich@bioelectromed.com [BioElectroMed Corp., 849 Mitten Rd., Suite 104, Burlingame, CA 94010 (United States)] [BioElectroMed Corp., 849 Mitten Rd., Suite 104, Burlingame, CA 94010 (United States); Tran, Kevin; Athos, Brian; Kreis, Mark; Nuccitelli, Pamela [BioElectroMed Corp., 849 Mitten Rd., Suite 104, Burlingame, CA 94010 (United States)] [BioElectroMed Corp., 849 Mitten Rd., Suite 104, Burlingame, CA 94010 (United States); Chang, Kris S.; Epstein, Ervin H. [The Children's Hospital Oakland Research Institute, Oakland, CA 94609 (United States)] [The Children's Hospital Oakland Research Institute, Oakland, CA 94609 (United States); Tang, Jean Y. [The Children's Hospital Oakland Research Institute, Oakland, CA 94609 (United States) [The Children's Hospital Oakland Research Institute, Oakland, CA 94609 (United States); Stanford University, Stanford, CA 94305 (United States)

2012-08-03

214

Membrane heterogeneity in murine stem cells  

SciTech Connect

The cell membrane's role in hemopoietic differentiation and regulation is increasingly evident. Certain fluorescent molecules interact with membrane components at the molecular level. The organic dye merocyanine 540(MCN) is such a tool for studying hemopoietic stem cell membranes at the molecular level. This paper present work involving biologic interactions of MCN with CLL (chronic lymphocytic leukemia) and hemopoietic cell membranes. The authors work indicates that MNC staining reveals membrane differences that accompany cellular transformation and differentiation. Changes in MNC fluorescene indicate that hemopoietic cells are heterogeneous with respect to membrane properties, and sensitivity to MNC photoinactivation reveals that membrane properties may change as a function of differentiative state between pluripotent and committed granulocytic precursor cells.

Myers, C.P.; Kerk, D.K.; Norman, A.

1980-01-01

215

Cyclosporine A induces apoptosis in murine tubular epithelial cells: Role of caspases  

Microsoft Academic Search

Cyclosporine A induces apoptosis in murine tubular epithelial cells: Role of caspases.BackgroundThe pathogenesis of cyclosporine A (CsA) nephrotoxicity has not been completely elucidated.MethodsThe ability of CsA to induce apoptosis in cultured murine tubular epithelial cells and its regulation by the cell microenvironment and inhibitors of caspases were studied.ResultsThis study found that CsA induces apoptotic death in murine proximal tubular epithelial

Alberto Ortiz; Corina Lorz; Marina Catalán; Arturo Ortiz; Santiago Coca; Jesus Egido

1998-01-01

216

Antiproliferative activity of lichen extracts on murine myeloma cells  

Microsoft Academic Search

In the present study we report some preliminary results concerning the evaluation of antiproliferative activity on murine\\u000a myeloma cells (P3X63-Ag8.653) of crude extracts of two common lichen species, Evernia prunastri and Xanthoria parietina.\\u000a \\u000a The results were evaluated by means of the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test, which\\u000a is commonly used to assess the activity of living cells through mitochondrial dehydrogenases. They

Doriana Triggiani; Donatella Ceccarelli; Antonio Tiezzi; Tommaso Pisani; Silvana Munzi; Carlo Gaggi; Stefano Loppi

2009-01-01

217

Allogeneic Cell Therapy for a Murine Mammary Carcinoma1  

Microsoft Academic Search

ABSTRACT The effect of allogeneic cell therapy on tumor,growth,was studied in a murine,model,of mammary,carcinoma,(4T1) as an experimental,model,of solid tumors in humans, i.v. inoculation of 4T1 (H-2d) cells into syngeneic mice [BALB\\/c or (BALB\\/cXC57BL\\/6)F,J (F,) carrying the H-2d histocom- patible antigens results in tumor,colonies in the lungs that finally cause the death of all of the mice. Sublet hüll \\\\ irradiated

Shoshana Morecki; Ellena Yacovlev; Adi Diab; Shimon Slavin

1998-01-01

218

Mesenchymal cells with leukocyte and lymphendothelial characteristics in murine embryos.  

PubMed

The development of lymphatic endothelial cells (LECs) from deep embryonic veins or mesenchymal lymphangioblasts is controversially discussed. Studies employing quail-chick grafting experiments have shown that various mesodermal compartments of the embryo possess lymphangiogenic potential, whereas studies on murine embryos have been in favor of a venous origin of LECs. We have investigated NMRI mice from embryonic day (ED) 9.5 to 13.5 with antibodies against the leukocyte marker CD45, the pan-endothelial marker CD31, and the lymphendothelial markers Prox1 and Lyve-1. Early signs of the development of lymphatics are the Lyve-1- and Prox1-positive segments of the jugular and vitelline veins. Then, lymph sacs, which are found in the jugular region of ED 11.5 mice, express Prox1, Lyve-1, and CD31. Furthermore, scattered cells positive for all of the four markers are present in the mesenchyme of the dermatomes and the mediastinum before lymphatic vessels are present in these regions. Their number increases during development. A gradient of increasing CD31 expression can be seen the closer the cells are located to the lymph sacs. Our studies provide evidence for the existence of scattered mesenchymal cells, which up-regulate lymphendothelial and down-regulate leukocyte characteristics when they integrate into growing murine lymphatics. Such stem cells may also be present in the human and may be the cell of origin in post-transplantation Kaposi sarcoma. PMID:16502417

Buttler, Kerstin; Kreysing, Alice; von Kaisenberg, Constantin S; Schweigerer, Lothar; Gale, Nick; Papoutsi, Maria; Wilting, Jörg

2006-06-01

219

Stromal cells participate in the murine esophageal mucosal injury response  

PubMed Central

We identified ?-smooth muscle actin (?-SMA)- and vimentin-expressing spindle-shaped esophageal mesenchymal cells in the adult and neonate murine esophageal lamina propria. We hypothesized that these esophageal mesenchymal cells express and secrete signaling and inflammatory mediators in response to injury. We established primary cultures of esophageal mesenchymal cells using mechanical and enzymatic digestion. We demonstrate that these primary cultures are nonhematopoietic, nonendothelial, stromal cells with myofibroblast-like features. These cells increase secretion of IL-6 in response to treatment with acidified media and IL-1?. They also increase bone morphogenetic protein (Bmp)-4 secretion in response to sonic hedgehog. The location of these cells and their biological functions demonstrate their potential role in regulating esophageal epithelial responses to injury and repair.

Binkley, Jana; Darwech, Isra; Swietlicki, Elzbieta; McDonald, Keely; Newberry, Rodney; Rubin, Deborah C.

2013-01-01

220

Packaging of Endogenous Retroviral Sequences in Retroviral Vectors Produced by Murine and Human Packaging Cells  

Microsoft Academic Search

Interaction of retrovirus vectors and endogenous retroviruses present in packaging cell lines and target cells may result in unwanted events, such as the formation of recombinant viruses and the mobilization of therapeu- tic vectors. Using sensitive reverse transcriptase PCR assays, we investigated human and murine gene therapy packaging cell lines for incorporation of endogenous retrovirus transcripts into murine leukemia virus

CLIVE PATIENCE; YASUHIRO TAKEUCHI; FRANCOIS-LOIC COSSET; ROBIN A. WEISS

1998-01-01

221

Estradiol Partially Recapitulates Murine Pituitary Cell Cycle Response to Pregnancy  

PubMed Central

Because pregnancy and estrogens both induce pituitary lactotroph hyperplasia, we assessed the expression of pituitary cell cycle regulators in two models of murine pituitary hyperplasia. Female mice were assessed during nonpregnancy, pregnancy, day of delivery, and postpartum. We also implanted estradiol (E2) pellets in female mice and studied them for 2.5 months. Pituitary weight in female mice increased 2-fold after E2 administration and 1.4-fold at day of delivery, compared with placebo-treated or nonpregnant females. Pituitary proliferation, as assessed by proliferating cell nuclear antigen and/or Ki-67 staining, increased dramatically during both mid-late pregnancy and E2 administration, and lactotroph hyperplasia was also observed. Pregnancy induced pituitary cell cycle proliferative and inhibitory responses at the G1/S checkpoint. Differential cell cycle regulator expression included cyclin-dependent kinase inhibitors, p21Cip1, p27Kip1, and cyclin D1. Pituitary cell cycle responses to E2 administration partially recapitulated those effects observed at mid-late pregnancy, coincident with elevated circulating mouse E2, including increased expression of proliferating cell nuclear antigen, Ki-67, p15INK4b, and p21Cip1. Nuclear localization of pituitary p21Cip1 was demonstrated at mid-late pregnancy but not during E2 administration, suggesting a cell cycle inhibitory role for p21Cip1 in pregnancy, yet a possible proproliferative role during E2 administration. Most observed cell cycle protein alterations were reversed postpartum. Murine pituitary meets the demand for prolactin during lactation associated with induction of both cell proliferative and inhibitory pathways, mediated, at least partially, by estradiol.

Toledano, Yoel; Zonis, Svetlana; Ren, Song-Guang; Wawrowsky, Kolja; Chesnokova, Vera

2012-01-01

222

Sertoli-Leydig cell tumor  

MedlinePLUS

... Testicular cancer. In Goldman L, Schafer AI, eds. Cecil Medicine . 24th ed. Philadelphia, PA: Saunders Elsevier; 2011: ... Gynecologic cancers. In Goldman L, Schafer AI, eds. Cecil Medicine . 24th ed. Philadelphia, PA: Saunders Elsevier; 2011: ...

223

Monoclonal antibodies specific for murine CD2 reveal its presence on B as well as T cells.  

PubMed Central

Monoclonal antibodies specific for the murine CD2 antigen were identified by an efficient screening method utilizing murine CD2 cDNA transfectants. An unexpected expression of CD2 on murine B cells was revealed by immunofluorescence and immunoprecipitation studies with these monoclonal antibodies and by RNA blot analysis for the murine CD2 transcript. Images

Yagita, H; Nakamura, T; Karasuyama, H; Okumura, K

1989-01-01

224

Generation of mesenchymal stem cell lines from murine bone marrow.  

PubMed

Mesenchymal stem cells (MSC), because of their multipotency and ease of purification and amplification, are an ideal stem cell source for cell therapies. Bone-marrow-derived stem cells (BMSC) can be used to develop MSC-like immortalized cell lines with large proliferation and differentiation potentialities. Their immortalized status prevents the maintenance of MSC function and characters; this can be negated by modifying the isolation and maintenance protocol. Adult murine BMSC were isolated and maintained in media without additional growth factors together with passage-dependent reseeding following trypsinization. Cells maintained over 25 passages were considered as putative cell lines and characterized. The phenotypic and genotypic characteristics and multilineage differentiation potential of the cells were assessed by morphological, phenotypic, and molecular assays at various passages. The putative BMSC cell lines showed the characteristics of MSC and were able to maintain these characteristics, even after immortalization. The phenotypic data demonstrated difference among two cell lines; this was further validated by the difference in their multilineage differentiation potential following specific induction. More importantly, no changes were observed in the genotypic level in comparison with control cells, even after more than 50 passages. Our protocol thus advances the isolation and maintenance of BMSC and the development of putative BMSC cell lines that maintain characteristics of MSC, including multilineage differentiation potential, after more than 40 passages. PMID:22836234

Sreejit, P; Dilip, K B; Verma, R S

2012-10-01

225

Enzymatic Induced Unmasking of Fetal Antigens on Normal Murine Hemopoietic Stem Cell Surfaces.  

National Technical Information Service (NTIS)

Based upon information provided by indirect immunofluorescence and x-ray fluorescence, this investigation has demonstrated that fetal determinants are expressed upon the cell surface of adult cells, specifically; normal adult murine hemopoietic stem cells...

B. M. Sheinberg

1977-01-01

226

Isolation and Identification of Cancer StemLike Cells from Murine Melanoma Cell Lines  

Microsoft Academic Search

In current study, cancer stem-like cells in the murine melanoma B16F10 cells were investigated. CD phenotypes of the B16F10 cells were analyzed by flow cytometry, and the specific CD phenotype cells from the B16F10 cells were isolated by MACS. Then we used colony formation assay in soft agar media, the cell growth assay in serum-free culture media as well as

Jun Dou; Meng Pan; Ping Wen; Yating Li; Quan Tang; Lili Chu; Fengshu Zhao; Chuilian Jiang; Weihua Hu; Kai Hu; Ning Gu

2007-01-01

227

Catecholamines in murine bone marrow derived mast cells.  

PubMed

Cultured murine bone marrow derived mast cells (BMMC) were found to store high levels of dopamine (3753+/-844 pg/10(7) cells) and occasionally produce norepinephrine and epinephrine. The catecholamine synthesis inhibitor, alpha-methyl-para-tyrosine, decreased intracellular catecholamine concentrations, and activation with ionomycin stimulated dopamine release. Neither dopaminergic receptor antagonists nor exogenous dopamine < or =10 microM affected IL-3-induced cell proliferation. High exogenous dopamine (20-100 microM) decreased proliferation and increased apoptosis, and the anti-oxidant ascorbic acid prevented these effects. Increased expression of the anti-apoptotic factor Bcl-2 or loss of pro-apoptotic Bax expression attenuated dopamine-induced apoptosis, suggesting the apoptosis proceeds through a mitochondrial pathway. PMID:11585626

Freeman, J G; Ryan, J J; Shelburne, C P; Bailey, D P; Bouton, L A; Narasimhachari, N; Domen, J; Siméon, N; Couderc, F; Stewart, J K

2001-10-01

228

Murine norovirus transcytosis across an in vitro polarized murine intestinal epithelial monolayer is mediated by M-like cells.  

PubMed

Noroviruses (NoVs) are the causative agent of the vast majority of nonbacterial gastroenteritis worldwide. Due to the inability to culture human NoVs and the inability to orally infect a small animal model, little is known about the initial steps of viral entry. One particular step that is not understood is how NoVs breach the intestinal epithelial barrier. Murine NoV (MNV) is the only NoV that can be propagated in vitro by infecting murine macrophages and dendritic cells, making this virus an attractive model for studies of different aspects of NoV biology. Polarized murine intestinal epithelial mICcl2 cells were used to investigate how MNV interacts with and crosses the intestinal epithelium. In this in vitro model of the follicle-associated epithelium (FAE), MNV is transported across the polarized cell monolayer in the absence of viral replication or disruption of tight junctions by a distinct epithelial cell with microfold (M) cell properties. In addition to transporting MNV, these M-like cells also transcytose microbeads and express an IgA receptor. Interestingly, B myeloma cells cultured in the basolateral compartment underlying the epithelial monolayer did not alter the number of M-like cells but increased their transcytotic activity. Our data demonstrate that MNV can cross an intact intestinal epithelial monolayer in vitro by hijacking the M-like cells' intrinsic transcytotic pathway and suggest a potential mechanism for MNV entry into the host. PMID:24049163

Gonzalez-Hernandez, Mariam B; Liu, Thomas; Blanco, Luz P; Auble, Heather; Payne, Hilary C; Wobus, Christiane E

2013-12-01

229

Murine Norovirus Transcytosis across an In Vitro Polarized Murine Intestinal Epithelial Monolayer Is Mediated by M-Like Cells  

PubMed Central

Noroviruses (NoVs) are the causative agent of the vast majority of nonbacterial gastroenteritis worldwide. Due to the inability to culture human NoVs and the inability to orally infect a small animal model, little is known about the initial steps of viral entry. One particular step that is not understood is how NoVs breach the intestinal epithelial barrier. Murine NoV (MNV) is the only NoV that can be propagated in vitro by infecting murine macrophages and dendritic cells, making this virus an attractive model for studies of different aspects of NoV biology. Polarized murine intestinal epithelial mICcl2 cells were used to investigate how MNV interacts with and crosses the intestinal epithelium. In this in vitro model of the follicle-associated epithelium (FAE), MNV is transported across the polarized cell monolayer in the absence of viral replication or disruption of tight junctions by a distinct epithelial cell with microfold (M) cell properties. In addition to transporting MNV, these M-like cells also transcytose microbeads and express an IgA receptor. Interestingly, B myeloma cells cultured in the basolateral compartment underlying the epithelial monolayer did not alter the number of M-like cells but increased their transcytotic activity. Our data demonstrate that MNV can cross an intact intestinal epithelial monolayer in vitro by hijacking the M-like cells' intrinsic transcytotic pathway and suggest a potential mechanism for MNV entry into the host.

Gonzalez-Hernandez, Mariam B.; Liu, Thomas; Blanco, Luz P.; Auble, Heather; Payne, Hilary C.

2013-01-01

230

Effects of gold sodium thiomalate on murine spleen cells  

SciTech Connect

The effects of gold sodium thiomalate (GST) on Balb/c murine spleen cells were investigated using in vitro mitogen blastogenesis techniques. Initial culture of spleen cells in the presence of concanavalin A (Con A) and GST exhibited inhibition of /sup 3/H-thymidine uptake due to gold. Spleen cells depleted of monocytes/macrophages by carbonyl iron-ingestion and cultured with Con A and GST demonstrated biphasic effects by gold on thymidine uptake depending upon the Con A and GST concentrations. A final concentration of 2.5 ..mu..g/ml con A showed a decreased inhibition of blastogenesis by gold, with macrophage depleted cells as compared to intact spleen cells. While a final concentration of 0.5 ..mu..g/ml Con A showed an increased inhibition of blastogenesis by gold with macrophage depleted cells as compared to intact spleen cells. GST was found to be most effective at inhibiting proliferation when present at the initiation of culture. GST was also found to inhibit responsiveness of cells previously incubated with mitogen and its presence in the culture media was required for at least 24-36 hours to produce significant inhibition. These results indicate that the greatest effects of GST on blastogenesis occur during the initial steps of lymphocyte activation and that macrophages may not be the only cell of gold action.

Brownback, P.; Measel, J.

1986-03-01

231

Nitric Oxide-Mediated Tumoricidal Activity of Murine Microglial Cells12  

PubMed Central

Experimental metastases in the brain of mice are infiltrated by microglia, and parabiosis experiments of green fluorescent protein (GFP+) and GFP- mice revealed that these microglia are derived from circulating monocytes (GFP+, F4/80+, and CD68+). These findings raised the question as to whether microglia (specialized macrophages) possess tumoricidal activity. C8-B4 murine microglia cells were incubated in vitro in medium (control) or in medium containing both lipopolysaccharide and interferon-?. Control microglia were not tumoricidal against a number of murine and human tumor cells, whereas lipopolysaccharide/interferon-?-activated microglia lysed murine and human tumor cells by release of nitric oxide. Parallel experiments with murine peritoneal macrophages produced identical results. Neither activated microglia nor activated macrophages lysed nontumorigenic murine or human cells. Collectively, these data demonstrate that brain metastasis-associated microglia are derived from circulating mononuclear cells and exhibit selective and specific tumoricidal activity.

Brantley, Emily C; Guo, Lixia; Zhang, Chenyu; Lin, Qingtang; Yokoi, Kenji; Langley, Robert R; Kruzel, Ewa; Maya, Marva; Kim, Seung Wook; Kim, Sun-Jin; Fan, Dominic; Fidler, Isaiah J

2010-01-01

232

Resistance to cyclopentenylcytosine in murine leukemia L1210 cells.  

PubMed

Cyclopentenyl cytosine (CPEC) exhibits oncological activity in murine and human tumor cells and has now entered Phase I clinical trials. Its mode of action as an antitumor agent appears to be inhibition by its triphosphate (CPEC-TP) of CTP synthase, the enzyme which converts UTP to CTP. In an attempt to elucidate the mechanism of resistance to CPEC, a murine leukemia cell line resistant to CPEC (L1210/CPEC) was developed by N-methyl-N-nitro-N-nitrosoguanidine-induced mutagenesis and subsequent selection by cultivation of the L1210 cells in the presence of 2 microM CPEC. Resistant clones were maintained in CPEC-free medium for 6 generations before biochemical studies were performed. The resistant clone selected for further studies was approximately 13,000-fold less sensitive to growth inhibition by CPEC than the parental cells, and the concentration of CPEC required to deplete CTP in the resistant cells was 50-fold higher than in the sensitive cells. A comparison of the kinetic properties of CTP synthase from sensitive and resistant cells indicated alteration in the properties of the enzyme from the latter; the median inhibitory concentration for CPEC-TP increased from 2 to 14 microM, Km for UTP decreased from 126 to 50 microM, and Vmax increased 12-fold from 0.2 to 2.3 nmol/mg/min. Northern blot analyses of polyadenylated RNA from the resistant and sensitive cells indicated a 3-fold increase in transcripts of the CTP synthase gene in the resistant line. Consistent with these alterations in the properties of the enzyme, the resistant cells exhibited significantly expanded CTP and dCTP pools (4- 5-fold) when compared with the sensitive cells. No change was observed, however, in the properties of uridine-cytidine kinase, the enzyme responsible for the initial phosphorylation of CPEC; despite this, however, cellular uptake of CPEC was greatly decreased, and phosphorylation of CPEC and its incorporation into RNA were 10-fold less than in the parental cells. These latter observations are most readily explained by feedback inhibition by the increased CTP levels of the resistant cells of uridine-cytidine kinase and/or of the membrane transport process used for initial entry of CPEC. PMID:7694793

Zhang, H; Cooney, D A; Zhang, M H; Ahluwalia, G; Ford, H; Johns, D G

1993-12-01

233

Enhanced proliferation of bone marrow mesenchymal stem cells by co-culture with TM4 mouse Sertoli cells: involvement of the EGF/PI3K/AKT pathway.  

PubMed

Bone marrow mesenchymal stem cells (BM-MSCs) are considered as a promising option in the field of regenerative medicine and tissue engineering. However, little is known about how TM4 mouse Sertoli cells, which are known to enhance stem cells proliferation in co-culture, may influence the proliferation of BM-MSCs and which signaling pathways are involved in. To address these questions, an in vitro transwell system was used. We found that TM4 cells could produce soluble factors which enhanced the growth of BM-MSCs without inhibiting the multipotency. Furthermore, cell cycle analysis showed that co-culture with the TM4 cells accelerated the progress of BM-MSCs from the G1 to the S phase. The expression of the phospho-akt, mdm2, as well as pho-CDC2, and cyclin D1 were markedly upregulated in co-cultured BM-MSCs. The observed promoting effect was significantly inhibited by the administration of the PI3K/AKT inhibitor, LY294002. Among the various growth factors produced by TM4 cells, the epithelial growth factor (EGF) stimulated the proliferation of the BM-MSCs more significantly compared with the other growth factors examined in this study. Neutralization of EGF via a blocking antibody significantly limited the promoting growth effect in BM-MSCs. These results suggest that TM4 cells provide a favorable in vitro environment for BM-MSCs growth and imply the involvement of the EGF/PI3K/AKT pathway. PMID:24748323

Tian, Huan; Guo, Meijin; Zhuang, Yingping; Chu, Ju; Zhang, Siliang

2014-08-01

234

Functional expression of SK channels in murine detrusor PDGFR?+ cells  

PubMed Central

We sought to characterize molecular expression and ionic conductances in a novel population of interstitial cells (PDGFR?+ cells) in murine bladder to determine how these cells might participate in regulation of detrusor excitability. PDGFR?+ cells and smooth muscle cells (SMCs) were isolated from detrusor muscles of PDGFR?+/eGFP and smMHC/Cre/eGFP mice and sorted by FACS. PDGFR?+ cells were highly enriched in Pdgfra (12 fold vs. unsorted cell) and minimally positive for Mhc (SMC marker), Kit (ICC marker) and Pgp9.5 (neuronal marker). SK3 was dominantly expressed in PDGFR?+ cells in comparison to SMCs. ?Slo (BK marker) was more highly expressed in SMCs. SK3 protein was observed in PDGFR?+ cells by immunohistochemistry but could not be resolved in SMCs. Depolarization evoked voltage-dependent Ca2+ currents in SMCs, but inward current conductances were not activated in PDGFR?+ cells under the same conditions. PDGFR?+ cells displayed spontaneous transient outward currents (STOCs) at potentials positive to ?60 mV that were inhibited by apamin. SK channel modulators, CyPPA and SKA-31, induced significant hyperpolarization of PDGFR?+ cells and activated SK currents under voltage clamp. Similar responses were not resolved in SMCs at physiological potentials. Single channel measurements confirmed the presence of functional SK3 channels (i.e. single channel conductance of 10 pS and sensitivity to intracellular Ca2+) in PDGFR?+ cells. The apamin-sensitive stabilizing factor regulating detrusor excitability is likely to be due to the expression of SK3 channels in PDGFR?+ cells because SK agonists failed to elicit resolvable currents and hyperpolarization in SMCs at physiological potentials.

Lee, Haeyeong; Koh, Byoung H; Peri, Lauren E; Sanders, Kenton M; Koh, Sang Don

2013-01-01

235

Functional expression of SK channels in murine detrusor PDGFR+ cells.  

PubMed

We sought to characterize molecular expression and ionic conductances in a novel population of interstitial cells (PDGFR?(+) cells) in murine bladder to determine how these cells might participate in regulation of detrusor excitability. PDGFR?(+) cells and smooth muscle cells (SMCs) were isolated from detrusor muscles of PDGFR?(+)/eGFP and smMHC/Cre/eGFP mice and sorted by FACS. PDGFR?(+) cells were highly enriched in Pdgfra (12 fold vs. unsorted cell) and minimally positive for Mhc (SMC marker), Kit (ICC marker) and Pgp9.5 (neuronal marker). SK3 was dominantly expressed in PDGFR?(+) cells in comparison to SMCs. ?Slo (BK marker) was more highly expressed in SMCs. SK3 protein was observed in PDGFR?(+) cells by immunohistochemistry but could not be resolved in SMCs. Depolarization evoked voltage-dependent Ca(2+) currents in SMCs, but inward current conductances were not activated in PDGFR?(+) cells under the same conditions. PDGFR?(+) cells displayed spontaneous transient outward currents (STOCs) at potentials positive to -60 mV that were inhibited by apamin. SK channel modulators, CyPPA and SKA-31, induced significant hyperpolarization of PDGFR?(+) cells and activated SK currents under voltage clamp. Similar responses were not resolved in SMCs at physiological potentials. Single channel measurements confirmed the presence of functional SK3 channels (i.e. single channel conductance of 10 pS and sensitivity to intracellular Ca(2+)) in PDGFR?(+) cells. The apamin-sensitive stabilizing factor regulating detrusor excitability is likely to be due to the expression of SK3 channels in PDGFR?(+) cells because SK agonists failed to elicit resolvable currents and hyperpolarization in SMCs at physiological potentials. PMID:23148317

Lee, Haeyeong; Koh, Byoung H; Peri, Lauren E; Sanders, Kenton M; Koh, Sang Don

2013-01-15

236

Effects of trichostatins on differentiation of murine erythroleukemia cells  

SciTech Connect

The fungistatic antibiotics trichostatins (TS) A and C were isolated from culture broth of Streptomyces platensis No. 145 and were found to be potent inducers of differentiation in murine erythroleukemia (Friend and RV133) cells at concentrations of 1.5 X 10(-8) M for TSA and 5 X 10(-7) M for TSC. Differentiation induced by TS was cooperatively enhanced by UV irradiation but not by treatment with dimethyl sulfoxide. This enhanced activity was completely inhibited by adding cycloheximide to the culture medium 2 h after exposure to TS, suggesting that TS are dimethyl sulfoxide-type inducers of erythroid differentiation. No inhibitory effect of TS was observed on macromolecular synthesis in cultured cells.

Yoshida, M.; Nomura, S.; Beppu, T.

1987-07-15

237

Chemotactic and Chemokinetic Activities of Stem Cell Factor on Murine Hematopoietic Progenitor Cells  

Microsoft Academic Search

We investigated the effects of stem cell factor (SCF) on the migration of murine bone marrow hematopoietic progenitor cells (HPC) in vitro using a modification of the checkerboard assay. Chemotactic and chemokinetic activities of SCF on HPC were evaluated by the numbers of HPC migrated on positive and negative gradients of SCF, respectively. On both positive and negative gradients of

Nobuo Okumura; Kohichiro Tsuji; Yasuhiro Ebihara; Nobukuni Sawai; Kenichi Koike; Atsushi Komiyama; Tatsutoshi Nakahata

1996-01-01

238

Cell cycle dependency of murine cytomegalovirus replication in synchronized 3T3 cells.  

PubMed Central

Synchronized murine 3T3 cells have been used to investigate the possible dependency of murine cytomegalovirus replication upon the cell cycle. The normal latent period of 12 h characteristic of asynchronous 3T3 cells was protracted to more than 24 h after an early G1 infection in synchronous cells. In this case viral progeny were not detected until after the initiation of the host S-phase. Cells maintained in the G1 phase did not replicate virus. This failure could not be explained by a decrease in virus penetration but was apparently due to a requirement for an event associated with the host S-phase. Thymidine-induced inhibition of cell cycle traverse also blocked virus replication. Viral DNA synthesis did not initiate until after the initiation of host DNA. In contrast, herpes simplex virus type 1 replicated in 3T3 cells independently of the cell cycle.

Muller, M T; Hudson, J B

1977-01-01

239

A study of Sertoli-spermatid tubulobulbar complexes in selected mammals.  

PubMed

Tubulobulbar complexes (TBCs) were found in nine mammalian species (opossum, vole, guinea-pig, mouse, hamster, rabbit, dog, monkey and human) primarily originating from the plasma membrane overlying the acrosome of late spermatids. Fewer complexes (4--10) were noted in these species than has been previously reported for the rat (up to 24). TBCs were not seen emanating from round spermatids or those elongated spermatids located within the deep recesses of the Sertoli cell, but they appeared as the spermatids came to reside much closer to the tubular lumen in preparation for release. TBCs developed in areas deficient or lacking in Sertoli filaments and endoplasmic reticulum (ectoplasmic specialization). In general their structural configuration was similar to that shown in the rat, although minor differences were noted. Fine fibrils were observed connecting the distal portion of the spermatid tube with the Sertoli plasma membrane forming a bristle-coated pit. The length of TBCs from most species studied was 1--2 micrometers, whereas those of the opossum extended 6--8 micrometers into an apical Sertoli process. TBCs were degraded within the Sertoli cell by its lysosomes prior to sperm release, and for most species there was evidence indicating that formation of more than one generation of TBCs occurred. As sperm release approached, TBCs formed preferentially from the leading edge of spermatids with spatulate heads. The Sertoli cell gradually withdrew from around the spermatid head until only the tip of the head was embedded within the Sertoli cell. This region of contact frequently demonstrated TBCs. The proposed functions of TBCs are reviewed and discussed in light of these findings from other species. PMID:6998046

Russell, L D; Malone, J P

1980-01-01

240

An ES-Like Pluripotent State in FGF-Dependent Murine iPS cells  

Microsoft Academic Search

Recent data demonstrates that stem cells can exist in two morphologically, molecularly and functionally distinct pluripotent states; a naïve LIF-dependent pluripotent state which is represented by murine embryonic stem cells (mESCs) and an FGF-dependent primed pluripotent state represented by murine and rat epiblast stem cells (EpiSCs). We find that derivation of induced pluripotent stem cells (iPSCs) under EpiSC culture conditions

Bruno di Stefano; Christa Buecker; Federica Ungaro; Alessandro Prigione; Hsu-Hsin Chen; Maaike Welling; Maureen Eijpe; Gustavo Mostoslavsky; Paul Tesar; James Adjaye; Niels Geijsen; Vania Broccoli; Martin Pera

2010-01-01

241

P2X7 receptor activation induces cell death and microparticle release in murine erythroleukemia cells  

Microsoft Academic Search

Extracellular ATP induces cation fluxes in and impairs the growth of murine erythroleukemia (MEL) cells in a manner characteristic of the purinergic P2X7 receptor, however the presence of P2X7 in these cells is unknown. This study investigated whether MEL cells express functional P2X7. RT-PCR, immunoblotting and immunofluorescence staining demonstrated the presence of P2X7 in MEL cells. Cytofluorometric measurements demonstrated that

Patrick Constantinescu; Bin Wang; Kati Kovacevic; Iman Jalilian; Giel J. C. G. M. Bosman; James S. Wiley; Ronald Sluyter

2010-01-01

242

Chinese medicinal herbs inhibit growth of murine renal cell carcinoma.  

PubMed

Tumors are known to produce factors suppressing immune functions. We previously showed that a murine renal cell carcinoma (Renca) suppressed macrophage function in vitro and that this suppression was abolished by co-incubation with extracts of two Chinese medicinal herbs. We now report that these phytochemicals are capable of inhibiting growth of Renca in vivo. BALB/c mice were transplanted intraperitoneally (IP) with 1-2 x 10(5) Renca cells. One day after tumor transplant, mice were randomized into two groups. One group was treated IP, daily for 10 days, with 100 microliters of phytochemicals containing 500 micrograms each of Astragalus membranaceus and Ligustrum lucidum, while the other group received saline as controls. A cure rate of 57% was obtained with these phytochemicals when the initial tumor load was 2 x 10(5), and 100% when the initial tumor load was 1 x 10(5). Additional experiments were performed to investigate the mechanisms involved in this protection. Splenic macrophages from tumor-bearing mice were shown to have depressed chemiluminescent oxidative burst activity, and this depression was restored with phytochemical treatment. Splenocytes from mice transplanted with Renca responded less favorably to interleukin-2 (IL-2) in generating lymphokine-activated killer (LAK) cells; again this depression was restored with phytochemical treatment. Our data suggest that these phytochemicals may have exerted their antitumor effects via augmentation of phagocyte and LAK cell activities. PMID:7812364

Lau, B H; Ruckle, H C; Botolazzo, T; Lui, P D

1994-01-01

243

Chimaerism and erythroid marker expression after microinjection of human acute myeloid leukaemia cells into murine blastocysts  

Microsoft Academic Search

It has been suggested that the embryonic microenvironment can control the survival and the transformed phenotype of tumour cells. Here, we addressed the hypothesis that the murine embryonic microenvironment can induce the differentiation of human tumour cells. To examine such interactions, we injected human leukaemic cells into preimplantation murine blastocysts at embryonic day 3.5 of gestation (E3.5). Microinjection of human

Michael Dürr; Friedrich Harder; Angela Merkel; Gesine Bug; Reinhard Henschler; Albrecht M Müller

2003-01-01

244

Establishment and characterization of a new murine cell line (SR4987) derived from marrow stromal cells  

Microsoft Academic Search

A new murine cell line designated as SR-4987 was established by treating a long-term bone marrow culture with the supernatant\\u000a from Y-1 cells which actively produce viral C-particles (MuLV). The line showed a fibrolbast-like morphology and its mesodermal\\u000a origin was confirmed by immunocytochemical staining. Flow cytometric analysis of DNA index evidenced a tetraploid number of\\u000a chromosomes whereas cell cycle analysis

Augusto Pessina; Elisabetta Mineo; Maria Grazia Neri; Laura Gribaldo; Robert Colombi; Paolo Brambilla; Gintaras Zaleskis

1992-01-01

245

Involvement of a chromatin modifier in response to mono-(2-ethylhexyl) phthalate (MEHP)-induced Sertoli cell injury: probably an indirect action via the regulation of NF?B/FasL circuitry.  

PubMed

The Fas/FasL signaling pathway, controlled by nuclear factor-?B (NF?B) at the transcriptional level, is critical for triggering germ cell apoptosis in response to mono-(2-ethylhexyl) phthalate (MEHP)-induced Sertoli cell (SC) injury, but the exact regulation mechanism remain unknown. Here, we discovered that expression level of Metastasis associated protein 1 (MTA1), a component of the Mi-2/nucleosome remodeling and deacetylase complex, was upregulated in SCs during the early recovery after MEHP exposure. This expression change was in line with the dynamic changes in germ cell apoptosis in response to MEHP treatment. Furthermore, a knockdown of MTA1 by RNAi in SCs was found to impair the MEHP-induced early activation of NF?B pathway and abolish the recruitment of NF?B onto FasL promoter, which consequently diminished the MEHP-triggered FasL induction. Considering that Fas/FasL is a well characterized apoptosis initiating signaling during SCs injury, our results point to a potential "switch on" effect of MTA1, which may govern the activation of NF?B/FasL cascade in MEHP-insulted SCs. Overall, the MTA1/NF?B/FasL circuit may serve as an important defensive/repairing mechanism to help to control the germ cell quality after SCs injury. PMID:24120950

Chen, Shiwei; Dong, Yushu; Xu, Chun; Jiang, Liming; Chen, Yongjie; Jiang, Cheng; Hou, Wugang; Li, Wei

2013-11-01

246

Echinacea purpurea extracts modulate murine dendritic cell fate and function.  

PubMed

Echinacea is a top-selling herbal remedy that purportedly acts as an immunostimulant. However, the specific immunomodulatory effects of Echinacea remain to be elucidated. We focused on defining the effects of Echinacea purpurea extracts in dendritic cells (DCs), which generate innate and adaptive immune responses. We hypothesized that E. purpurea extracts would enhance murine bone marrow-derived DC (BMDC) activation leading to increased immune responses. The fate and function of DCs from C57Bl/6 mice was evaluated following 48h exposure to E. purpurea root and leaf extracts. Flow cytometry revealed that the polysaccharide-rich root extract increased the expression of MHC class II, CD86, and CD54 surface biomarkers whereas the alkylamide-rich leaf extract inhibited expression of these molecules. Production of IL-6 and TNF-alpha increased in a concentration-dependent manner with exposure to the root, but not leaf, extract. In contrast, the leaf but not root extract inhibited the enzymatic activity of cyclooxygenase-2. While both extracts decreased the uptake of ovalbumin by BMDCs, the leaf but not root extract inhibited the antigen-specific activation of naïve CD4(+) T cells from OT II/Thy1.1 mice. Collectively, these results suggest that E. purpurea can be immunostimulatory, immunosuppressive, and/or anti-inflammatory depending on the portion of the plant and extraction method. PMID:20149833

Benson, Jenna M; Pokorny, Amanda J; Rhule, Ava; Wenner, Cynthia A; Kandhi, Vamsikrishna; Cech, Nadja B; Shepherd, David M

2010-05-01

247

Echinacea pupurea extracts modulate murine dendritic cell fate and function  

PubMed Central

Echinacea is a top-selling herbal remedy that purportedly acts as an immunostimulant. However, the specific immunomodulatory effects of Echinacea remain to be elucidated. We focused on defining the effects of Echinacea purpurea extracts in dendritic cells (DCs), which generate innate and adaptive immune responses. We hypothesized that E. purpurea extracts would enhance murine bone marrow-derived DC (BMDC) activation leading to increased immune responses. The fate and function of DCs from C57Bl/6 mice was evaluated following 48 h exposure to E. purpurea root and leaf extracts. Flow cytometry revealed that the polysaccharide-rich root extract increased the expression of MHC class II, CD86, and CD54 surface biomarkers whereas the alkylamide-rich leaf extract inhibited expression of these molecules. Production of IL-6 and TNF-? increased in a concentration-dependent manner with exposure to the root, but not leaf, extract. In contrast, the leaf but not root extract inhibited the enzymatic activity of cyclooxygenase-2. While both extracts decreased the uptake of ovalbumin by BMDCs, the leaf but not root extract inhibited the antigen-specific activation of naïve CD4+ T cells from OT II/Thy1.1 mice. Collectively, these results suggest that E. purpurea can be immunostimulatory, immunosuppressive, and/or anti-inflammatory depending on the portion of the plant and extraction method.

Benson, Jenna M.; Pokorny, Amanda J.; Rhule, Ava; Wenner, Cynthia A.; Kandhi, Vamsikrishna; Cech, Nadja B.; Shepherd, David M.

2010-01-01

248

Vitamin d controls murine and human plasmacytoid dendritic cell function.  

PubMed

Topical application of the vitamin D (VitD) analog calcipotriol is a highly effective standard treatment modality of psoriatic skin lesions. However, the immune modulatory effects of the treatment are incompletely understood. VitD is well known to induce tolerogenic responses in conventional dendritic cells (cDCs). Plasmacytoid DCs (pDCs) comprise a specialized, naturally occurring DC subset known to be important in autoimmune diseases including psoriasis. pDCs from the blood rapidly infiltrate psoriatic skin and are key to the initiation of the immune-mediated pathogenesis of the disease. We now demonstrate that pDCs express various proteins of the VitD receptor (VDR) pathway, including the VitD-metabolizing enzymes Cyp27B1 and Cyp24A1, and that VDR is transcriptionally active in pDCs. Moreover, VitD impairs the capacity of murine and human pDCs to induce T-cell proliferation and secretion of the T-helper 1 cytokine IFN?. The inhibitory effect of VitD is dependent on the expression of the VDR in the DCs. This study demonstrates that VitD signaling can act as a natural inhibitory mechanism on both cDCs and pDCs, which may instigate the development of VitD-based therapeutic applications for psoriasis and other inflammatory skin diseases. PMID:24352045

Karthaus, Nina; van Spriel, Annemiek B; Looman, Maaike W G; Chen, Shuo; Spilgies, Lisanne M; Lieben, Liesbet; Carmeliet, Geert; Ansems, Marleen; Adema, Gosse J

2014-05-01

249

Identification of the Gross cell surface antigen associated with murine leukemia virus-infected cells.  

PubMed Central

The Gross cell surface antigen (GCSA) is produced by cells that are either exogenously infected with murine leukemia virus (MuLV) or are expressing endogenous MuLV genomes. In immune precipitation assays, GCSA was resolved into two serologically distinct 85,000- and 95,000-dalton viral proteins. These antigenic components are glycosylated forms of the polyprotein precursors of the MuLV internal structural proteins. Images

Ledbetter, J; Nowinski, R C

1977-01-01

250

Tellurite-induced oxidative stress leads to cell death of murine hepatocarcinoma cells  

Microsoft Academic Search

Data regarding tellurium (Te) toxicity are scarce. Studies on its metabolism, performed mainly in bacteria, underline a major\\u000a role of reactive oxygen species (ROS). We investigated whether tellurite undergoes redox cycling leading to ROS formation\\u000a and cancer cell death. The murine hepatocarcinoma Transplantable Liver Tumor (TLT) cells were challenged with tellurite either\\u000a in the presence or in the absence of

Juan M. Sandoval; Philippe Levêque; Bernard Gallez; Claudio C. Vásquez; Pedro Buc Calderon

2010-01-01

251

Constancy of cell buoyant density for cultured murine cells  

SciTech Connect

The relationship between cell cycle and cell density was determined for three different lines of mouse cells by equilbrium centrifugation of suspension cultures. The mean cell densities of the three lines differed significantly, with values of 1.0622, 1.0678, 1.0540 gm/ml for 70Z/3, S 107, and ABE 8, respectively. However the density distributions within each of the three lines were indistinguishable, with an average coefficient of variation about 5% of the mean reduced density. Quantitative DNA analysis of the cells separated by density showed that the proportion of cells in G1, S, and G2 + M phase of the cell cycle changed very little or not at all with cell density. In addition, cells separated by size using velocity sedimentation had the same means and distributions of densities. These results indicate that there is little or no changes in cell density as the cells traverse the life cycle and that buoyant density appears to be a constant property of a cell type. 12 references, 4 figures, 2 tables.

Loken, M.R.; Kubitschek, H.E.

1984-01-01

252

Isolation of a mesenchymal cell population from murine dermis that contains progenitors of multiple cell lineages  

PubMed Central

The skin contains two known subpopulations of stem cells/epidermal progenitors, a basal keratinocyte population found in the interfollicular epithelium and cells residing in the bulge region of the hair follicle. The major role of the interfollicular basal keratinocyte population may be epidermal renewal, whereas the bulge population may only be activated and recruited to form a cutaneous epithelium in case of trauma. Utilizing three-dimensional cultures of murine skin under stress conditions in which only reserve epithelial cells would be expected to survive and expand, we demonstrate that a mesenchymal population resident in neonatal murine dermis has the unique potential to develop an epidermis in vitro. In monolayer culture, this dermal subpopulation has long-term survival capabilities in restricted serum, and inducible capacity to evolve into multiple cell lineages, both epithelial and mesenchymal, depending on culture conditions. When grafted subcutaneously, this dermal subpopulation gave rise to fusiform structures, reminiscent of disorganized muscle, that stained positive for smooth muscle actin and desmin; on typical epidermal grafts, abundant melanocytes appeared throughout the dermis that were not associated with hair follicles. The multipotential cells can be repeatedly isolated from neonatal murine dermis by a sequence of differential centrifugation and selective culture conditions. These results suggest that progenitors capable of epidermal differentiation exist in the mesenchymal compartment of an abundant tissue source and may have a function in mesenchymal-epithelial transition upon insult; moreover these cells could be available in sufficient quantities for lineage determination or tissue engineering applications.

Crigler, Lauren; Kazhanie, Amita; Yoon, Tae-Jin; Zakhari, Julia; Anders, Joanna; Taylor, Barbara; Virador, Victoria M.

2007-01-01

253

Developmental changes of CD34 expression by murine hematopoietic stem cells  

Microsoft Academic Search

ObjectiveIt has been reported that fetal murine hematopoietic stem cells are CD34+, whereas adult stem cells are CD34?. We sought to delineate the developmental changes of CD34 expression by hematopoietic stem cells and carried out systematic analysis of long-term engrafting cells in the bone marrow and\\/or blood of perinatal, juvenile, and adult mice.

Tatsuya Ito; Fumihito Tajima; Makio Ogawa

2000-01-01

254

Multiple effects of dendritic cell depletion on murine norovirus infection.  

PubMed

Dendritic cells (DCs) are permissive to murine norovirus (MNV) infection in vitro and in vivo. However, their roles during infection in vivo are not well defined. To determine the role of DCs during infection, conventional DCs were depleted from CD11c-DTR mice and infected with a persistent MNV strain. Viral titres in the intestine and secondary lymphoid organs were determined at early time points during infection, and anti-MNV antibody responses were analysed later during infection. Depletion of conventional DCs resulted in increased viral loads in intestinal tissues, impaired generation of antibody responses, and a failure of MNV to efficiently infect lymphoid tissues. These data suggest that DCs play multiple roles in MNV pathogenesis, in both innate immunity and the efficient generation of adaptive immune responses against MNV, as well as by promoting the dissemination of MNV to secondary lymphoid tissues. This is the first study to probe the roles of DCs in controlling and/or facilitating a norovirus infection in vivo and provides the basis for further studies aimed at defining mechanisms by which DCs control MNV replication and promote viral dissemination. PMID:23636823

Elftman, Michael D; Gonzalez-Hernandez, Mariam B; Kamada, Nobuhiko; Perkins, Cheryl; Henderson, Kenneth S; Núñez, Gabriel; Wobus, Christiane E

2013-08-01

255

Murine granulated metrial gland cells are susceptible to Chlamydia psittaci infection in vivo.  

PubMed Central

Granulated metrial gland (GMG) cells are the most numerous lymphoid cells in the uteroplacental unit in rodent pregnancy. In an experimental murine model of abortion-causing infection, we have studied the responses of GMG cells to Chlamydia psittaci. Chlamydial inclusions have been found within GMG cells, both in apparently healthy cells and in cells with degenerative changes. Establishing the existence of GMG cells infected by C. psittaci opens a new and interesting chapter in the study of these cells.

Sanchez, J; Buendia, A J; Salinas, J; Bernabe, A; Rodolakis, A; Stewart, I J

1996-01-01

256

Liver sinusoidal endothelial cells promote lymphohematopoietic differentiation from murine embryonic stem cells: Role of soluble factors.  

PubMed

Liver sinusoid endothelial cells (LSEC) constitute an in vitro and in vivo microenvironment for the proliferation and differentiation of hematopoietic stem cells (HSC). Previously, we have shown that LSEC support the survival and growth of murine embryonic stem cells (ESC). In this study, we investigated the capacity of LSEC to promote hematopoietic differentiation from the murine ESC cell line, CGR8. Undifferentiated ESC were cultured on LSEC monolayers in the absence of exogenous cytokines. After 10 and 20 days, cells were harvested and examined by their morphology, phenotype and capacity of hematopoietic colony formation. Microscopic observation of LSEC/ESC cocultures showed the presence of cobblestone areas formation, which indicates active hematopoiesis. Morphological analysis of cell from these foci showed the presence of hematopoietic cells at different stages of differentiation. Cells expressing B lymphoid markers (B220 and CD19) were detected by flow cytometry, and clonogenic assays showed the formation of CFU-pre B colonies. Similar results were observed when ESC were cultured with LSEC conditioned media. Myeloid precursors were also detected by the presence of CFU-GM colonies and cells expressing myeloid markers. These results indicate that LSEC provided an in vitro microenvironment mainly for B cell development, but also myeloid differentiation from ESC. Coculture of ESC with LSEC may constitute a very powerful tool to study the mechanisms involved in B cell generation from ESC. PMID:24892988

Silva-Cote, Ingrid; Cardier, Jose E

2014-09-01

257

Expression of coagulation factors from murine induced pluripotent stem cell-derived liver cells.  

PubMed

A protocol to differentiate liver cells from induced pluripotent stem (iPS) cells is being established. However, the ability of these differentiated iPS cells to express liver-specific proteins, such as coagulation cascade and related factors, has yet to be assessed. This study evaluated whether liver-like populations differentiated from murine iPS cells gain the ability to produce coagulation-related factors. Following differentiation of murine iPS cells into hematopoietic-like and liver-like embryoid bodies, we assessed gene expression profiles for coagulation-related markers, including fibrinogen, factors II, V, VII, VIII, IX, X, XI, XII, and XIII?, protein C, protein S, antithrombin, plasminogen, von Willebrand factor, and ADAMTS13 by real-time reverse transcription PCR. Liver-like embryoid bodies demonstrated strong expression levels of nearly all the coagulation-related genes assessed, compared with undifferentiated iPS cells and hematopoietic-like embryoid bodies. We also confirmed efficient translation and secretion of fibrinogen and albumin (hepatocyte-specific marker proteins) into the conditioned medium by these differentiated cells, suggesting successful differentiation of iPS cells into the liver lineage. These findings suggest that iPS cells can be differentiated into liver-like populations that express coagulation-related factors. Liver-like embryoid bodies may provide a source for cell-based therapies directed toward liver diseases, including coagulation factor deficiencies in the future. PMID:21415711

Kasuda, Shogo; Tatsumi, Kohei; Sakurai, Yoshihiko; Kato, Junko; Taminishi, Sanae; Takeda, Tomohiro; Ohashi, Kazuo; Okano, Teruo; Hatake, Katsuhiko; Shima, Midori

2011-06-01

258

Damage and recovery of mouse testis after 1000 r acute localized x-irradiation, with reference to restitution cells, sertoli cell increase, and type a. Spermatogonial recovery  

Microsoft Academic Search

After 1000 r of acute localized irradiation many spermatocytes are ; blocked in first metaphase. Many restitution nuclei similar to those seen after ; colchicine treatment form between the first and the tenth day after irradiation, ; the maximum being observed at 6 days. These restituted cells often drift toward ; the basement membrane. In agreement with previous investigators we

B. R. Nebel; C. J. Murphy

1960-01-01

259

Selection and extended growth of murine epidermal stem cells in culture.  

PubMed

Continuously renewing epithelia contain small undifferentiated stem cells capable of self-renewal and maintenance of the differentiating cell population. In murine epidermis stem cells have been identified as label-retaining cells (LRCs) by long-term retention of tritiated thymidine or BrdU. It has been suggested that epidermal stem cells adhere to basement membranes through differential expression of specific integrins. To determine whether we could use a specific integrin to enrich for murine epidermal stem cells, we tested adherence of LRCs to several substrates. Regardless of the substrate used, approximately 10% of total basal cells and 100% of LRCs adhered in 10 min. In our medium specifically formulated for murine keratinocytes, rapidly adherent stem cells formed large colonies and could be used to form a structurally complete epidermis in organotypic culture. They showed a fivefold greater transient transfection efficiency than total basal cells, and when individual adherent cells were transduced with a retroviral vector, they formed large clones. Although these stem cells grew more slowly than the total basal cell population, they could be subcultured more times. Our results indicate that murine epidermal stem cells can be selected by rapid attachment to a substrate, but not by one specific integrin, and that they can be expanded in culture if the appropriate conditions are maintained. PMID:9770361

Bickenbach, J R; Chism, E

1998-10-10

260

TLX1 (HOX11) Immortalization of Embryonic Stem Cell-Derived and Primary Murine Hematopoietic Progenitors  

PubMed Central

The ability to generate genetically engineered cell lines is of great experimental value. They provide a renewable source of material that may be suitable for biochemical analyses, chromatin immunoprecipitation assays, structure-function studies, gene function assignment, and transcription factor target gene identification. This unit describes protocols for TLX1 (HOX11)-mediated immortalization of murine hematopoietic progenitors derived from in vitro differentiated murine embryonic stem cells, or from primary mouse fetal liver or bone marrow. A wide variety of hematopoietic cell types have been immortalized using these procedures including erythroid, megakaryocytic, monocytic, myelocytic, and multipotential cell types. These lines are typically cytokine dependent for their survival and growth.

Hawley, Robert G.; Hawley, Teresa S.; Cantor, Alan B.

2009-01-01

261

Erythroid Cell Differentiation: Murine Erythroleukemia Cell Variant with Unique Pattern of Induction by Polar Compounds  

Microsoft Academic Search

The murine-virus-infected erythroleukemia cell system provides an opportunity to examine regulatory mechanisms controlling cytodifferentiation. A cloned cell line (DR10c3) resistant to the erythropoiesis-inducing effect of dimethylsulfoxide (Me2SO) was isolated from the Me2SO-sensitive line DS19. DR10c3 is characterized as follows: (1) the uptake of [3H]Me2SO is similar to that in DS19; (2) cell growth with and without Me2SO is similar to

Yoshiro Ohta; Masao Tanaka; Masaaki Terada; Orlando J. Miller; Arthur Bank; Paul A. Marks; Richard A. Rifkind

1976-01-01

262

Homophilic peptide inhibits growth of malignant murine and human B cells.  

PubMed

A homophilic peptide from the T15 plasmacytoma inhibits growth of murine and human B cell tumors. This finding confirms the hypothesis that B cell malignancies are driven by a self-binding epitope in the B cell receoptor (BCR) proposed as the pathogenesis of chronic lymphocytic leukemia (CLL). PMID:24328748

Kohler, Heinz; Bryan, Ann Jay

2013-12-01

263

Involvement of a chromatin modifier in response to mono-(2-ethylhexyl) phthalate (MEHP)-induced Sertoli cell injury: Probably an indirect action via the regulation of NF?B/FasL circuitry  

SciTech Connect

Highlights: •MTA1 expression is upregulated in SCs upon MEHP treatment. •Knockdown of MTA1 in SCs impairs the MEHP-induced NF?B signaling activation. •Knockdown of MTA1 inhibits recruitment of NF?B onto FasL promoter in MEHP-treated SCs. -- Abstract: The Fas/FasL signaling pathway, controlled by nuclear factor-?B (NF?B) at the transcriptional level, is critical for triggering germ cell apoptosis in response to mono-(2-ethylhexyl) phthalate (MEHP)-induced Sertoli cell (SC) injury, but the exact regulation mechanism remain unknown. Here, we discovered that expression level of Metastasis associated protein 1 (MTA1), a component of the Mi-2/nucleosome remodeling and deacetylase complex, was upregulated in SCs during the early recovery after MEHP exposure. This expression change was in line with the dynamic changes in germ cell apoptosis in response to MEHP treatment. Furthermore, a knockdown of MTA1 by RNAi in SCs was found to impair the MEHP-induced early activation of NF?B pathway and abolish the recruitment of NF?B onto FasL promoter, which consequently diminished the MEHP-triggered FasL induction. Considering that Fas/FasL is a well characterized apoptosis initiating signaling during SCs injury, our results point to a potential “switch on” effect of MTA1, which may govern the activation of NF?B/FasL cascade in MEHP-insulted SCs. Overall, the MTA1/NF?B/FasL circuit may serve as an important defensive/repairing mechanism to help to control the germ cell quality after SCs injury.

Chen, Shiwei [Department of Urology, 174th Hospital of PLA, Fujian 361001 (China)] [Department of Urology, 174th Hospital of PLA, Fujian 361001 (China); Dong, Yushu [Department of Neurosurgery, 463rd Hospital of PLA, Shenyang 110042 (China)] [Department of Neurosurgery, 463rd Hospital of PLA, Shenyang 110042 (China); Xu, Chun; Jiang, Liming; Chen, Yongjie; Jiang, Cheng [Department of Urology, 174th Hospital of PLA, Fujian 361001 (China)] [Department of Urology, 174th Hospital of PLA, Fujian 361001 (China); Hou, Wugang, E-mail: gangwuhou@163.com [Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032 (China)] [Department of Anesthesiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032 (China); Li, Wei, E-mail: liweipepeyato@163.com [Department of Human Anatomy, Histology and Embryology, Fourth Military Medical University, Xi’an 710032 (China)] [Department of Human Anatomy, Histology and Embryology, Fourth Military Medical University, Xi’an 710032 (China)

2013-11-01

264

Pseudolaric Acid B Induced Cell Cycle Arrest, Autophagy and Senescence in Murine Fibrosarcoma L929 Cell  

PubMed Central

Objective: PAB induced various cancer cell apoptosis, cell cycle arrest and senescence. But in cell line murine fibrosarcoma L929, PAB did not induce apoptosis, but autophagy, therefore it was thought by us as a good model to research the relationship of cell cycle arrest, autophagy and senescence bypass apoptosis. Methods: Inhibitory ratio was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Phase contrast microscopy visualized cell morphology. Hoechst 33258 staining for nuclear change, propidium iodode (PI) staining for cell cycle, monodansylcadaverine (MDC) staining for autophagy, and rodanmine 123 staining for mitochondrial membrane potential (MMP) were measured by fluorescence microscopy or flowcytometry. Apoptosis was determined by DNA ladder test. Protein kinase C (PKC) activity was detected by PKC assay kit. SA-?-galactosidase assay was used to detect senescence. Protein expression was examined by western blot. Results: PAB inhibited L929 cell growth in time-and dose-dependent manner. At 12 h, 80 ?mol/L PAB induced obvious mitotic arrest; at 24 h, PAB began to induce autophagy; at 36 h, cell-treated with PAB slip into G1 cell cycle; and 3 d PAB induced senescence. In time sequence PAB induced firstly cell cycle arrest, then autophagy, then slippage into G1 phase, lastly senescence. Senescent cells had high level of autophagy, inhibiting autophagy led to apoptosis, and no senescence. PAB activated PKC activity to induce cell cycle arrest, autophagy and senescence, inhibiting PKC activity suppressed cell cycle arrest, autophagy and senescence. Conclusion: PAB induced cell cycle arrest, autophagy and senescence in murine fibrosarcoma L929 cell through PKC.

hua Yu, Jing; yu Liu, Chun; bin Zheng, Gui; Zhang, Li Ying; hui Yan, Ming; yan Zhang, Wen; ying Meng, Xian; fang Yu, Xiao

2013-01-01

265

Plasmacytoid dendritic cells promote rotavirus-induced human and murine B cell responses  

PubMed Central

B cell–dependent immunity to rotavirus, an important intestinal pathogen, plays a significant role in viral clearance and protects against reinfection. Human in vitro and murine in vivo models of rotavirus infection were used to delineate the role of primary plasmacytoid DCs (pDCs) in initiating B cell responses. Human pDCs were necessary and sufficient for B cell activation induced by rotavirus. Type I IFN recognition by B cells was essential for rotavirus-mediated B cell activation in vitro and murine pDCs and IFN-?/?–mediated B cell activation after in vivo intestinal rotavirus infection. Furthermore, rotavirus-specific serum and mucosal antibody responses were defective in mice lacking functional pDCs at the time of infection. These data demonstrate that optimal B cell activation and virus-specific antibody secretion following mucosal infection were a direct result of pDC-derived type I IFN. Importantly, viral shedding significantly increased in pDC-deficient mice, suggesting that pDC-dependent antibody production influences viral clearance. Thus, mucosal pDCs critically influence the course of rotavirus infection through rotavirus recognition and subsequent IFN production and display powerful adjuvant properties to initiate and enhance humoral immunity.

Deal, Emily M.; Lahl, Katharina; Narvaez, Carlos F.; Butcher, Eugene C.; Greenberg, Harry B.

2013-01-01

266

Potent Inhibition of Jun?n Virus Infection by Interferon in Murine Cells  

PubMed Central

The new world arenavirus Junín virus (JUNV) is the causative agent of Argentine hemorrhagic fever, a lethal human infectious disease. Adult laboratory mice are generally resistant to peripheral infection by JUNV. The mechanism underlying the mouse resistance to JUNV infection is largely unknown. We have reported that interferon receptor knockout mice succumb to JUNV infection, indicating the critical role of interferon in restricting JUNV infection in mice. Here we report that the pathogenic and vaccine strains of JUNV were highly sensitive to interferon in murine primary cells. Treatment with low concentrations of interferon abrogated viral NP protein expression in murine cells. The replication of both JUNVs was enhanced in IRF3/IRF7 deficient cells. In addition, the vaccine strain of JUNV displayed impaired growth in primary murine cells. Our data suggested a direct and potent role of host interferon response in restricting JUNV replication in mice. The defect in viral growth for vaccine JUNV might also partially explain its attenuation in mice.

Huang, Cheng; Walker, Aida G.; Grant, Ashley M.; Kolokoltsova, Olga A.; Yun, Nadezhda E.; Seregin, Alexey V.; Paessler, Slobodan

2014-01-01

267

Differentiated phenotypes of primary murine alveolar epithelial cells and their susceptibility to infection by respiratory viruses.  

PubMed

Severe respiratory viral infections are associated with spread to the alveoli of the lungs. There are multiple murine models of severe respiratory viral infections that have been used to identify viral and host factors that contribute to disease severity. Primary cultures of murine alveolar epithelial cells provide a robust in vitro model to perform mechanistic studies that can be correlated with in vivo studies to identify cell type-specific factors that contribute to pathology within the alveoli of the lung during viral infection. In this study, we established an in vitro model to compare the responses of type I (ATI) and type II (ATII) alveolar epithelial cells to infection by respiratory viruses used in murine models: mouse-adapted severe acute respiratory syndrome-associated coronavirus (SARS-CoV, v2163), murine coronavirus MHV-1, and influenza A (H1N1) virus, strain PR8. Murine alveolar cells cultured to maintain an ATII cell phenotype, determined by expression of LBP180, were susceptible to infection by all three viruses. In contrast, ATII cells that were cultured to trans-differentiate into an ATI-like cell phenotype were susceptible to MHV-1 and PR8, but not mouse-adapted SARS-CoV. Epithelial cells produce cytokines in response to viral infections, thereby activating immune responses. Thus, virus-induced cytokine expression was quantified in ATI and ATII cells. Both cell types had increased expression of IL-1? mRNA upon viral infection, though at different levels. While MHV-1 and PR8 induced expression of a number of shared cytokines in ATI cells, there were several cytokines whose expression was induced uniquely by MHV-1 infection. In summary, ATI and ATII cells exhibited differential susceptibilities and cytokine responses to infection by respiratory viruses. This in vitro model will be critical for future studies to determine the roles of these specialized cell types in the pathogenesis of respiratory viral infection. PMID:23639425

Kebaabetswe, Lemme P; Haick, Anoria K; Miura, Tanya A

2013-08-01

268

Cytokine Modulation of Murine Stem Cell Engraftment: The Role of Adherence to Plastic Surfaces  

Microsoft Academic Search

Murine marrow stem cells acquire an engraftment defect when cultured for 48 hours in cytokines, whereas the number of progenitor\\u000a cells expands. Stem or progenitor cells have been noted to adhere to various surfaces, including plastic. Despite vigorous\\u000a harvesting by cell scraping, the possibility existed that cytokines might induce selective adhesion of the rare engraftable\\u000a stem cells to plastic surfaces.We

Stefan O. Peters; Houri K. Habibian; Peter J. Quesenberry

2002-01-01

269

Neutrophil and B Cell Expansion in Mice That Lack the Murine IL8 Receptor Homolog  

Microsoft Academic Search

Interleukin-8 (IL-8) is a proinflammatory cytokine that specifically attracts and activates human neutrophils. A murine gene with a high degree of homology to the two known human IL-8 receptors was cloned and then deleted from the mouse genome by homologous recombination in embryonic stem (ES) cells. These mice, although outwardly healthy, had lymphadenopathy, resulting from an increase in B cells,

Grace Cacalano; James Lee; Kristine Kikly; Ann M. Ryan; Sharon Pitts-Meek; Bruce Hultgren; William I. Wood; Mark W. Moore

1994-01-01

270

Expression of authentic substance P receptors in murine and human dendritic cells  

Microsoft Academic Search

Recent studies from our laboratory have shown that substance P can elicit transcription factor activation in dendritic cells. In the present study, we extend these findings by demonstrating the presence of authentic substance P (NK-1) receptors on both normal murine and human dendritic cells. Specifically, we demonstrate the presence of mRNA encoding NK-1 tachykinin receptors and have utilized specific antibodies

Ian Marriott; Kenneth L. Bost

2001-01-01

271

Glutathione peroxidase-1 is required for self-renewal of murine embryonic stem cells.  

PubMed

Embryonic stem (ES) cells are pluripotent cells that are capable of giving rise to any type of cells in the body and possess unlimited self-renewal potential. However, the exact regulatory mechanisms that govern the self-renewal ability of ES cells remain elusive. To understand the immediate early events during ES cell differentiation, we performed a proteomics study and analyzed the proteomic difference in murine ES cells before and after a 6-h spontaneous differentiation. We found that the expression level of glutathione peroxidase-1 (GPx-1), an antioxidant enzyme, is dramatically decreased upon the differentiation. Both knockdown of GPx-1 expression with shRNA and inhibiting GPx-1 activity by inhibitor led to the differentiation of ES cells. Furthermore, we showed that during early differentiation, the quick degradation of GPx-1 was mediated by proteasome. Thus, our data indicated that GPx-1 is a key regulator of self-renewal of murine embryonic stem cells. PMID:24802396

Wang, Qian-Yi; Liu, Zhao-Shan; Wang, Jie; Wang, Hong-Xia; Li, Ang; Yang, Yang; Wang, Xin-Zheng; Zhao, Yong-Qiang; Han, Qiu-Ying; Cai, Hong; Liang, Bing; Song, Nan; Li, Wei-Hua; Li, Tao

2014-06-13

272

Isolation and characterization of side population cells in the postpartum murine endometrium.  

PubMed

Endometrium is a highly active organ that is periodically remodeled during the life span. Previous studies have indicated the presence of an adult stem or progenitor cell population in this tissue. In this study, side population (SP) cells were isolated from the endometrium of postpartum murine uterus but not from the endometrium of a uterus undergoing a normal estrus cycle. Phenotype analysis showed that SP cells were negative for hematopoietic, endothelial, and mesenchymal stem cell (MSC) markers, but they expressed stem cell antigen 1 (Sca-1) and c-kit at various degrees. Side population cell is a heterogeneous population of endometrial stem/progenitor cells that have colony-forming capacity. They were found to reside in quiescence in the stroma but not in the luminal epithelium. These data suggest that, like other tissues and organs, the murine endometrium also contains SP cells. Their specific role in the regeneration of the endometrium warrants further study. PMID:20581350

Hu, Fei-Fei; Jing Xu; Cui, Yu-Gui; Qian, Xiao-Qiao; Mao, Yun-Dong; Liao, Lian-Ming; Liu, Jia-Yin

2010-07-01

273

Genetic Ablation of Phosphatidylinositol Transfer Protein Function in Murine Embryonic Stem Cells  

PubMed Central

Phosphatidylinositol transfer proteins (PITPs) regulate the interface between signal transduction, membrane-trafficking, and lipid metabolic pathways in eukaryotic cells. The best characterized mammalian PITPs are PITP? and PITP?, two highly homologous proteins that are encoded by distinct genes. Insights into PITP? and PITP? function in mammalian systems have been gleaned exclusively from cell-free or permeabilized cell reconstitution and resolution studies. Herein, we report for the first time the use of genetic approaches to directly address the physiological functions of PITP? and PITP? in murine cells. Contrary to expectations, we find that ablation of PITP? function in murine cells fails to compromise growth and has no significant consequence for bulk phospholipid metabolism. Moreover, the data show that PITP? does not play an obvious role in any of the cellular activities where it has been reconstituted as an essential stimulatory factor. These activities include protein trafficking through the constitutive secretory pathway, endocytic pathway function, biogenesis of mast cell dense core secretory granules, and the agonist-induced fusion of dense core secretory granules to the mast cell plasma membrane. Finally, the data demonstrate that PITP?-deficient cells not only retain their responsiveness to bulk growth factor stimulation but also retain their pluripotency. In contrast, we were unable to evict both PITP? alleles from murine cells and show that PITP? deficiency results in catastrophic failure early in murine embryonic development. We suggest that PITP? is an essential housekeeping PITP in murine cells, whereas PITP? plays a far more specialized function in mammals than that indicated by in vitro systems that show PITP dependence.

Alb, James G.; Phillips, Scott E.; Rostand, Kathleen; Cui, Xiaoxia; Pinxteren, Jef; Cotlin, Laura; Manning, Timothy; Guo, Shuling; York, John D.; Sontheimer, Harald; Collawn, James F.; Bankaitis, Vytas A.

2002-01-01

274

Production of Immune Interferon by an Interleukin 2Independent Murine T Cell Line  

Microsoft Academic Search

An interleukin 2-independent murine T cell line (BFS) was isolated that produced immune interferon after stimulation with phorbol 12-myristate 13-acetate. The BFS cell line did not produce detectable levels of interleukin 1, interleukin 2, B cell growth factor, macrophage-granulocyte colony-stimulating factor, macrophage-activating factor, or T cell replacing factor. Maximal interferon was induced 48 hr after stimulation with phorbol myristate acetate

William R. Benjamin; Patricia S. Steeg; John J. Farrar

1982-01-01

275

Functional activity of murine CD34 +and CD34 ? hematopoietic stem cell populations  

Microsoft Academic Search

The transmembrane glycoprotein CD34 is expressed on human hematopoietic stem cells and committed progenitors in the bone marrow, and CD34-positive selection currently is used to isolate bone marrow repopulating cells in clinical transplantation protocols. Recently, CD34? hematopoietic stem cells were described in both humans and mice, and it was suggested that CD34+ murine bone marrow cells may lack long-term reconstituting

D. Scott Donnelly; Daniel Zelterman; Saul Sharkis; Diane S. Krause

1999-01-01

276

Th1\\/Th2-Regulated Expression of Arginase Isoforms in Murine Macrophages and Dendritic Cells  

Microsoft Academic Search

Activated murine macrophages metabolize arginine by two alternative pathways involving the enzymes inducible NO synthase (iNOS) or arginase. The balance between the two enzymes is competitively regulated by Th1 and Th2 T helper cells via their secreted cytokines: Th1 cells induce iNOS, whereas Th2 cells induce arginase. Whereas the role of macrophages expressing iNOS as inflammatory cells is well established,

Markus Munder; Klaus Eichmann; Francisco Centeno; German Soler; Manuel Modolell

277

The Ly49 family: regulation of cytokine production in murine NK cells  

Microsoft Academic Search

Most proteins encoded by members of the L.?y-49 gene family are class I-recognizing receptors on murine natural killer (NK) cells. Class I recognition by Ly-49 receptors usually results in inhibition of NK cell iysis of target cells. However, NK cells function not only in a lytic capacity, but also can mediate cy- tokine production. In this report we have demonstrated

J. A. Ortaldo; L. H. Mason; T. A. Gregorlo; J. Stoll; A. T. Winkter-Pickett

1997-01-01

278

Differentiation of subpopulations of human and murine hemopoietic stem cells by hypotonic lysis.  

PubMed Central

Both human and mouse bone marrow contain subpopulations of hemopoietic stem cells that greatly vary in their resistance to water exposure: The cells forming erythroid colonies or bursts in methyl cellulose in vitro are most sensitive to hypotonic conditions and are destroyed within 60 s in the hypotonic milieu. The murine pluripotent stem cells assayed by the spleen colony technique, as well as both murine and human myeloid stem cells assayed by the plasma clot diffusion chamber technique, displayed intermediate sensitivity and were nearly completely eliminated by 120 s of exposure to water. Both human and mouse bone marrow stem cells producing myeloid colonies in agar are most resistant to hypotonic conditions. The addition of monocyte-macrophages and lymphoid cells to water-exposed mouse bone marrow cell populations to compensate for losses did not restore either erythroid or myeloid colony formation.

Niskanen, E; Cline, M J

1980-01-01

279

Generation of a Large Number of Connective Tissue Type Mast Cells by Culture of Murine Fetal Skin Cells  

Microsoft Academic Search

We describe a novel culture system for generating large numbers of murine skin-associated mast cells and distinguish their characteristics from bone marrow-derived cultured mast cells. Culture of day 16 fetal skin single cell suspensions in the presence of interleukin-3 and stem cell factor allowed expansion and maturation of mast cells in the presence of stromal cells. The average yield of

Nobuo Yamada; Hironori Matsushima; Yutaka Tagaya; Shinji Shimada; Stephen I. Katz

2003-01-01

280

Cryoprotective effects of low-density lipoproteins, trehalose and soybean lecithin on murine spermatogonial stem cells.  

PubMed

Summary Spermatogonial stem cells (SSCs) have the ability to self-renew and offer a pathway for genetic engineering of the male germ line. Cryopreservation of SSCs has potential value for the treatment of male infertility, spermatogonial transplantation, and so on. In order to investigate the cryopreservation effects of different cryoprotectants on murine SSCs, 0.2 M of low-density lipoproteins (LDL), trehalose and soybean lecithin were added to the cryoprotective medium, respectively, and the murine SSCs were frozen at -80°C or -196°C. The results indicated that the optimal recovery rates of murine SSCs in the cryoprotective medium supplemented with LDL, trehalose and soybean lecithin were 92.53, 76.35 and 75.48% at -80°C, respectively. Compared with freezing at -196°C, the optimum temperature for improvement of recovery rates of frozen murine SSCs, cryopreservation in three different cryoprotectants at -80°C, were 17.11, 6.68 and 10.44% respectively. The recovery rates of murine SSCs in the cryoprotective medium supplemented with 0.2 M LDL were significantly higher than that of other cryoprotectants (P < 0.05). Moreover, the recovery rates were demonstrated to be greater at -80°C compared with at -196°C (P < 0.05). In conclusion, 0.2 M of LDL could significantly protect murine SSCs at -80°C. In the freezing-thawing process, LDL is responsible for the cryopreservation of murine SSCs because it can form a protective film at the surface of membranes. However, more research is needed to evaluate and understand the precise role of LDL during the freezing-thawing of SSCs. PMID:22974447

Wang, Peng; Li, Ying; Hu, Xiao-Chen; Cai, Xiao-Li; Hou, Li-Peng; Wang, Yan-Feng; Hu, Jian-Hong; Li, Qing-Wang; Suo, Li-Juan; Fan, Zhi-Guo; Zhang, Bo

2012-09-14

281

Dendritic Cells Are the Major Antigen Presenting Cells in Inflammatory Lesions of Murine Mycoplasma Respiratory Disease  

PubMed Central

Mycoplasmas cause chronic respiratory diseases in animals and humans, and to date, development of vaccines have been problematic. Using a murine model of mycoplasma pneumonia, lymphocyte responses, specifically T cells, were shown to confer protection as well as promote immunopathology in mycoplasma disease. Because T cells play such a critical role, it is important to define the role of antigen presenting cells (APC) as these cells may influence either exacerbation of mycoplasma disease pathogenesis or enhancement of protective immunity. The roles of APC, such as dendritic cells and/or macrophages, and their ability to modulate adaptive immunity in mycoplasma disease are currently unknown. Therefore, the purpose of this study was to identify individual pulmonary APC populations that may contribute to the activation of T cell responses during mycoplasma disease pathogenesis. The present study indeed demonstrates increasing numbers of CD11c? F4/80+ cells, which contain macrophages, and more mature/activated CD11c+ F4/80? cells, containing DC, in the lungs after infection. CD11c? F4/80+ macrophage-enriched cells and CD11c+ F4/80? dendritic cell-enriched populations showed different patterns of cytokine mRNA expression, supporting the idea that these cells have different impacts on immunity in response to infection. In fact, DC containing CD11c+ F4/80? cell populations from the lungs of infected mice were most capable of stimulating mycoplasma-specific CD4+ Th cell responses in vitro. In vivo, these CD11c+F4/80? cells were co-localized with CD4+ Th cells in inflammatory infiltrates in the lungs of mycoplasma-infected mice. Thus, CD11c+F4/80? dendritic cells appear to be the major APC population responsible for pulmonary T cell stimulation in mycoplasma-infected mice, and these dendritic cells likely contribute to responses impacting disease pathogenesis.

Sun, Xiangle; Jones, Harlan P.; Dobbs, Nicole; Bodhankar, Sheetal; Simecka, Jerry W.

2013-01-01

282

Dendritic cells are the major antigen presenting cells in inflammatory lesions of murine Mycoplasma respiratory disease.  

PubMed

Mycoplasmas cause chronic respiratory diseases in animals and humans, and to date, development of vaccines have been problematic. Using a murine model of mycoplasma pneumonia, lymphocyte responses, specifically T cells, were shown to confer protection as well as promote immunopathology in mycoplasma disease. Because T cells play such a critical role, it is important to define the role of antigen presenting cells (APC) as these cells may influence either exacerbation of mycoplasma disease pathogenesis or enhancement of protective immunity. The roles of APC, such as dendritic cells and/or macrophages, and their ability to modulate adaptive immunity in mycoplasma disease are currently unknown. Therefore, the purpose of this study was to identify individual pulmonary APC populations that may contribute to the activation of T cell responses during mycoplasma disease pathogenesis. The present study indeed demonstrates increasing numbers of CD11c(-) F4/80(+) cells, which contain macrophages, and more mature/activated CD11c(+) F4/80(-) cells, containing DC, in the lungs after infection. CD11c(-) F4/80(+) macrophage-enriched cells and CD11c(+) F4/80(-) dendritic cell-enriched populations showed different patterns of cytokine mRNA expression, supporting the idea that these cells have different impacts on immunity in response to infection. In fact, DC containing CD11c(+) F4/80(-) cell populations from the lungs of infected mice were most capable of stimulating mycoplasma-specific CD4(+) Th cell responses in vitro. In vivo, these CD11c(+)F4/80(-) cells were co-localized with CD4(+) Th cells in inflammatory infiltrates in the lungs of mycoplasma-infected mice. Thus, CD11c(+)F4/80(-) dendritic cells appear to be the major APC population responsible for pulmonary T cell stimulation in mycoplasma-infected mice, and these dendritic cells likely contribute to responses impacting disease pathogenesis. PMID:23390557

Sun, Xiangle; Jones, Harlan P; Dobbs, Nicole; Bodhankar, Sheetal; Simecka, Jerry W

2013-01-01

283

In vitro Stimulation of Murine Spleen Cells using a Microculture System and a Multiple Automated Sample Harvester.  

National Technical Information Service (NTIS)

A microculture system is described to study the in vitro properties of murine spleen cells. The use of a new multiple automated sample harvester enabled rapid multiple tests using relatively few cells. This new procedure was highly reproducible and dimini...

D. M. Strong A. A. Ahmed G. B. Thurman K. W. Sell

1973-01-01

284

P2X7 receptor activation induces cell death and microparticle release in murine erythroleukemia cells.  

PubMed

Extracellular ATP induces cation fluxes in and impairs the growth of murine erythroleukemia (MEL) cells in a manner characteristic of the purinergic P2X7 receptor, however the presence of P2X7 in these cells is unknown. This study investigated whether MEL cells express functional P2X7. RT-PCR, immunoblotting and immunofluorescence staining demonstrated the presence of P2X7 in MEL cells. Cytofluorometric measurements demonstrated that ATP induced ethidium+ uptake into MEL cells in a concentration-dependent fashion and with an EC(50) of approximately 154 microM. The most potent P2X7 agonist 2'- and 3'-0(4-benzoylbenzoyl) ATP, but not ADP or UTP, induced ethidium+ uptake. ATP-induced ethidium+ and YO-PRO-1(2+) uptake were impaired by the P2X7 antagonist, A-438079. A colourmetric assay demonstrated that ATP impaired MEL cell growth. A cytofluorometric assay showed that ATP induced MEL cell death and that this process was impaired by A-438079. Finally, cytofluorometric measurements of Annexin-V binding and bio-maleimide staining demonstrated that ATP could induce rapid phosphatidylserine exposure and microparticle release in MEL cells respectively, both of which were impaired by A-438079. These results demonstrate that MEL cells express functional P2X7, and indicate that activation of this receptor may be important in the death and release of microparticles from red blood cells in vivo. PMID:20529664

Constantinescu, Patrick; Wang, Bin; Kovacevic, Kati; Jalilian, Iman; Bosman, Giel J C G M; Wiley, James S; Sluyter, Ronald

2010-09-01

285

Identification of Replication Competent Murine Gammaretroviruses in Commonly Used Prostate Cancer Cell Lines  

Microsoft Academic Search

A newly discovered gammaretrovirus, termed XMRV, was recently reported to be present in the prostate cancer cell line CWR22Rv1. Using a combination of both immunohistochemistry with broadly-reactive murine leukemia virus (MLV) anti-sera and PCR, we determined if additional prostate cancer or other cell lines contain XMRV or MLV-related viruses. Our study included a total of 72 cell lines, which included

Karen Sandell Sfanos; Amanda L. Aloia; Jessica L. Hicks; David M. Esopi; Jared P. Steranka; Wei Shao; Silvia Sanchez-Martinez; Srinivasan Yegnasubramanian; Kathleen H. Burns; Alan Rein; Angelo M. De Marzo

2011-01-01

286

Mechanism of the melanogenesis stimulation activity of (?)-cubebin in murine B16 melanoma cells  

Microsoft Academic Search

(?)-Cubebin showed a melanogenesis stimulation activity in a concentration-dependent manner in murine B16 melanoma cells without any significant effects on cell proliferation. Tyrosinase activity was increased at 24–72h after addition of cubebin to B16 cells, and then intracellular melanin amount was increased at 48–96h after the treatment. The expression levels of tyrosinase were time-dependently enhanced after the treatment with cubebin.

Noriko Hirata; Shunsuke Naruto; Kenji Ohguchi; Yukihiro Akao; Yoshinori Nozawa; Munekazu Iinuma; Hideaki Matsuda

2007-01-01

287

Identification of tissue-specific vasculogenic cells originating from murine uterus  

Microsoft Academic Search

Endometrium is a highly regenerative adult tissue that undergoes repeated degeneration and regeneration following menarche.\\u000a Therefore, it is believed that endometrium contains stem and\\/or progenitor cells in order to compensate for the regeneration\\u000a of tissue components. We report here that stem-like cells having vasculogenic potential are present in the uterus. Enzymatically\\u000a extracted cells from murine uteri were characterized and fractionated

Narumi Onodera; Tetsuro Tamaki; Yoshinori Okada; Akira Akatsuka; Daisuke Aoki

2006-01-01

288

Comet assay in murine bone-marrow cell line (FDC-P2)  

Microsoft Academic Search

The comet assay, also known as the single cell gel electrophoresis (SCGE) assay, is a rapid, simple, visual and sensitive technique for measuring DNA damage in mammalian cells. In the present study, Methyl methanesulfonate (MMS), 4-Nitrosoquinoline-Oxide (4NQO), Cyclophosphamide (CPA), and Benzo(a)pyrene (BP)-induced DNA damage was assayed in vitro in a murine bone-marrow cell line (FDC-P2), with or without an activation

K. Oshida; E. Iwanaga; K. Miyamoto; Y. Miyamoto

2010-01-01

289

COMPARATIVE TOXICITY OF DIFFERENT EMISSION PARTICLES IN MURINE PULMONARY EPITHELIAL CELLS AND MACROPHAGES  

EPA Science Inventory

Comparative Toxicity of Different Emission Particles in Murine Pulmonary Epithelial Cells and Macrophages. T Stevens1, M Daniels2, P Singh2, M I Gilmour2. 1 UNC, Chapel Hill 27599 2Experimental Toxicology Division, NHEERL, RTP, NC 27711 Epidemiological studies have shown ...

290

Involvement of Retinal Neurons and Pigment Epithelial Cells in a Murine Model of Sandhoff Disease  

Microsoft Academic Search

Background\\/Aims: To investigate the effects of deficient degradation of glycolipids on the morphological appearance of all retinal cells in a murine model of GM2 gangliosidosis (Sandhoff disease). Methods: The morphological appearance of the retina in Sandhoff mice at symptomatic stages (3 and 4 months of age) was examined by immunohistochemistry, lectin histochemistry and electron microscopy. Results: Under a light microscope,

Kazunori Sango; Shoji Yamanaka; Kyoko Ajiki; Nobutaka Arai; Masahiko Takano

2008-01-01

291

GPBAR1/TGR5 Mediates Bile Acid-Induced Cytokine Expression in Murine Kupffer Cells  

PubMed Central

GPBAR1/TGR5 is a novel plasma membrane-bound G protein–coupled bile acid (BA) receptor. BAs are known to induce the expression of inflammatory cytokines in the liver with unknown mechanism. Here we show that without other external stimuli, TGR5 activation alone induced the expression of interleukin 1? (IL-1?) and tumor necrosis factor-? (TNF-?) in murine macrophage cell line RAW264.7 or murine Kupffer cells. The TGR5-mediated increase of pro-inflammatory cytokine expression was suppressed by JNK inhibition. Moreover, the induced pro-inflammatory cytokine expression in mouse liver by 1% cholic acid (CA) diet was blunted in JNK?/? mice. TGR5 activation by its ligands enhanced the phosphorylation levels, DNA-binding and trans-activities of c-Jun and ATF2 transcription factors. Finally, the induced pro-inflammatory cytokine expression in Kupffer cells by TGR5 activation correlated with the suppression of Cholesterol 7?-hydroxylase (Cyp7a1) expression in murine hepatocytes. These results suggest that TGR5 mediates the BA-induced pro-inflammatory cytokine production in murine Kupffer cells through JNK-dependent pathway. This novel role of TGR5 may correlate to the suppression of Cyp7a1 expression in hepatocytes and contribute to the delicate BA feedback regulation.

Lou, Guiyu; Ma, Xiaoxiao; Fu, Xianghui; Meng, Zhipeng; Zhang, Wenyu; Wang, Yan-Dong; Van Ness, Carl; Yu, Donna; Xu, Rongzhen; Huang, Wendong

2014-01-01

292

GPBAR1/TGR5 mediates bile acid-induced cytokine expression in murine Kupffer cells.  

PubMed

GPBAR1/TGR5 is a novel plasma membrane-bound G protein-coupled bile acid (BA) receptor. BAs are known to induce the expression of inflammatory cytokines in the liver with unknown mechanism. Here we show that without other external stimuli, TGR5 activation alone induced the expression of interleukin 1? (IL-1?) and tumor necrosis factor-? (TNF-?) in murine macrophage cell line RAW264.7 or murine Kupffer cells. The TGR5-mediated increase of pro-inflammatory cytokine expression was suppressed by JNK inhibition. Moreover, the induced pro-inflammatory cytokine expression in mouse liver by 1% cholic acid (CA) diet was blunted in JNK-/- mice. TGR5 activation by its ligands enhanced the phosphorylation levels, DNA-binding and trans-activities of c-Jun and ATF2 transcription factors. Finally, the induced pro-inflammatory cytokine expression in Kupffer cells by TGR5 activation correlated with the suppression of Cholesterol 7?-hydroxylase (Cyp7a1) expression in murine hepatocytes. These results suggest that TGR5 mediates the BA-induced pro-inflammatory cytokine production in murine Kupffer cells through JNK-dependent pathway. This novel role of TGR5 may correlate to the suppression of Cyp7a1 expression in hepatocytes and contribute to the delicate BA feedback regulation. PMID:24755711

Lou, Guiyu; Ma, Xiaoxiao; Fu, Xianghui; Meng, Zhipeng; Zhang, Wenyu; Wang, Yan-Dong; Van Ness, Carl; Yu, Donna; Xu, Rongzhen; Huang, Wendong

2014-01-01

293

Characterization of human mast cells developed in vitro from fetal liver cells cocultured with murine 3T3 fibroblasts.  

PubMed

Cocultures of dispersed human fetal liver cells with murine Swiss 3T3 fibroblasts resulted in the development of human mast cells after 1 to 4 weeks of culture. Mast cells were detected by immunohistochemistry using a murine monoclonal anti-tryptase antibody, before metachromasia appeared with toluidine blue. When subjected to double immunohistochemistry using murine monoclonal anti-chymase and anti-tryptase antibodies, 94% +/- 10% (SD) of the mast cells seen at day 30 of culture were of the MCT type. These results contrast with those obtained with human mast cells derived from cord blood mononuclear cells cocultured with murine 3T3 fibroblasts which are comprised of substantially greater numbers of MCTC cells, averaging 48% +/- 31% (SD) at day 30 of culture. Mast cells developed in vitro from fetal liver cells or cord blood mononuclear cells contained similar amounts (+/- SD) of histamine (0.9 +/- 0.5 pg/cell and 1.1 +/- 1 pg/cell, respectively) and tryptase (1.7 +/- 0.4 pg/cell and 1.9 +/- 1.2 pg/cell, respectively) on day 30 of culture. Fetal-liver-derived mast cells from a 30-day-old culture were identified by immunoelectron microscopy using gold-labelled antitryptase antibody. Typically, these mast cells appeared immature as they had large nuclear to cytoplasmic ratio and a small number of ill-formed cytoplasmic granules. For both fetal-liver- and cord-blood-derived mast cells, there was no evidence of conversion of the MCT type into the MCTC type provided by this study. These results suggest that commitment to develop as an MCT or MCTC type of mast cell may have occurred in mast cell precursors present in fetal liver and cord blood mononuclear cells, prior to granulation. PMID:1398760

Irani, A A; Craig, S S; Nilsson, G; Ishizaka, T; Schwartz, L B

1992-09-01

294

SPECIFIC MURINE B-CELL ACTIVATION BY SYNTHETIC SINGLE- AND DOUBLE-STRANDED POLYNUCLEOTIDES  

PubMed Central

The synthetic single- and double-stranded polynucleotides, poly I, poly C, and poly I·C, were shown to induce thymidine incorporation in six inbred strains of murine spleen cells. This stimulation was shown to be secondary to B-cell activation and not due to contamination of the polynucleotides with bacterial lipopolysaccharide (LPS). The ability of poly I·C to act as a B-cell mitogen, in addition to its behavior as a thymic-independent antigen, suggested that these two phenomena may be related. The similarity of the molecular structure of poly I·C to LPS, a material which also acts as a thymic-independent antigen and a B-cell mitogen, supports the hypothesis that the polyvalent nature of these materials accounts for their functional interaction with murine B cells.

Scher, Irwin; Strong, Douglas M.; Ahmed, Aftab; Knudsen, Richard C.; Sell, Kenneth W.

1973-01-01

295

Novel Murine Dendritic Cell Lines: A Powerful Auxiliary Tool for Dendritic Cell Research  

PubMed Central

Research in vitro facilitates discovery, screening, and pilot experiments, often preceding research in vivo. Several technical difficulties render Dendritic Cell (DC) research particularly challenging, including the low frequency of DC in vivo, thorough isolation requirements, and the vulnerability of DC ex vivo. Critically, there is not as yet a widely accepted human or murine DC line and in vitro systems of DC research are limited. In this study, we report the generation of new murine DC lines, named MutuDC, originating from cultures of splenic CD8? conventional DC (cDC) tumors. By direct comparison to normal WT splenic cDC subsets, we describe the phenotypic and functional features of the MutuDC lines and show that they have retained all the major features of their natural counterpart in vivo, the splenic CD8? cDC. These features include expression of surface markers Clec9A, DEC205, and CD24, positive response to TLR3 and TLR9 but not TLR7 stimuli, secretion of cytokines, and chemokines upon activation, as well as cross-presentation capacity. In addition to the close resemblance to normal splenic CD8? cDC, a major advantage is the ease of derivation and maintenance of the MutuDC lines, using standard culture medium and conditions, importantly without adding supplementary growth factors or maturation-inducing stimuli to the medium. Furthermore, genetically modified MutuDC lines have been successfully obtained either by lentiviral transduction or by culture of DC tumors originating from genetically modified mice. In view of the current lack of stable and functional DC lines, these novel murine DC lines have the potential to serve as an important auxiliary tool for DC research.

Fuertes Marraco, Silvia A.; Grosjean, Frederic; Duval, Anais; Rosa, Muriel; Lavanchy, Christine; Ashok, Devika; Haller, Sergio; Otten, Luc A.; Steiner, Quynh-Giao; Descombes, Patrick; Luber, Christian A.; Meissner, Felix; Mann, Matthias; Szeles, Lajos; Reith, Walter; Acha-Orbea, Hans

2012-01-01

296

Novel murine dendritic cell lines: a powerful auxiliary tool for dendritic cell research.  

PubMed

Research in vitro facilitates discovery, screening, and pilot experiments, often preceding research in vivo. Several technical difficulties render Dendritic Cell (DC) research particularly challenging, including the low frequency of DC in vivo, thorough isolation requirements, and the vulnerability of DC ex vivo. Critically, there is not as yet a widely accepted human or murine DC line and in vitro systems of DC research are limited. In this study, we report the generation of new murine DC lines, named MutuDC, originating from cultures of splenic CD8? conventional DC (cDC) tumors. By direct comparison to normal WT splenic cDC subsets, we describe the phenotypic and functional features of the MutuDC lines and show that they have retained all the major features of their natural counterpart in vivo, the splenic CD8? cDC. These features include expression of surface markers Clec9A, DEC205, and CD24, positive response to TLR3 and TLR9 but not TLR7 stimuli, secretion of cytokines, and chemokines upon activation, as well as cross-presentation capacity. In addition to the close resemblance to normal splenic CD8? cDC, a major advantage is the ease of derivation and maintenance of the MutuDC lines, using standard culture medium and conditions, importantly without adding supplementary growth factors or maturation-inducing stimuli to the medium. Furthermore, genetically modified MutuDC lines have been successfully obtained either by lentiviral transduction or by culture of DC tumors originating from genetically modified mice. In view of the current lack of stable and functional DC lines, these novel murine DC lines have the potential to serve as an important auxiliary tool for DC research. PMID:23162549

Fuertes Marraco, Silvia A; Grosjean, Frédéric; Duval, Anaïs; Rosa, Muriel; Lavanchy, Christine; Ashok, Devika; Haller, Sergio; Otten, Luc A; Steiner, Quynh-Giao; Descombes, Patrick; Luber, Christian A; Meissner, Felix; Mann, Matthias; Szeles, Lajos; Reith, Walter; Acha-Orbea, Hans

2012-01-01

297

Activation of murine lung mast cells by the adenosine A3 receptor.  

PubMed

Adenosine has been implicated to play a role in asthma in part through its ability to influence mediator release from mast cells. Most physiological roles of adenosine are mediated through adenosine receptors; however, the mechanisms by which adenosine influences mediator release from lung mast cells are not understood. We established primary murine lung mast cell cultures and used real-time RT-PCR and immunofluorescence to demonstrate that the A(2A), A(2B), and A(3) adenosine receptors are expressed on murine lung mast cells. Studies using selective adenosine receptor agonists and antagonists suggested that activation of A(3) receptors could induce mast cell histamine release in association with increases in intracellular Ca(2+) that were mediated through G(i) and phosphoinositide 3-kinase signaling pathways. The function of A(3) receptors in vivo was tested by exposing mice to the A(3) receptor agonist, IB-MECA. Nebulized IB-MECA directly induced lung mast cell degranulation in wild-type mice while having no effect in A(3) receptor knockout mice. Furthermore, studies using adenosine deaminase knockout mice suggested that elevated endogenous adenosine induced lung mast cell degranulation by engaging A(3) receptors. These results demonstrate that the A(3) adenosine receptor plays an important role in adenosine-mediated murine lung mast cell degranulation. PMID:12817016

Zhong, Hongyan; Shlykov, Sergiy G; Molina, Jose G; Sanborn, Barbara M; Jacobson, Marlene A; Tilley, Stephen L; Blackburn, Michael R

2003-07-01

298

Diabetes, insulin-mediated glucose metabolism and Sertoli/blood-testis barrier function  

PubMed Central

Blood testis barrier (BTB) is one of the tightest blood-barriers controlling the entry of substances into the intratubular fluid. Diabetes Mellitus (DM) is an epidemic metabolic disease concurrent with falling fertility rates, which provokes severe detrimental BTB alterations. It induces testicular alterations, disrupting the metabolic cooperation between the cellular constituents of BTB, with dramatic consequences on sperm quality and fertility. As Sertoli cells are involved in the regulation of spermatogenesis, providing nutritional support for germ cells, any metabolic alteration in these cells derived from DM may be responsible for spermatogenesis disruption, playing a crucial role in fertility/subfertility associated with this pathology. These cells have a glucose sensing machinery that reacts to hormonal fluctuations and several mechanisms to counteract hyper/hypoglycemic events. The role of DM on Sertoli/BTB glucose metabolism dynamics and the metabolic molecular mechanisms through which DM and insulin deregulation alter its functioning, affecting male reproductive potential will be discussed.

Alves, Marco G.; Martins, Ana D.; Cavaco, Jose E.; Socorro, Silvia; Oliveira, Pedro F.

2013-01-01

299

Influence of Murine Mesenchymal Stem Cells on Proliferation, Phenotype, Vitality, and Cytotoxicity of Murine Cytokine-Induced Killer Cells in Coculture  

PubMed Central

Stimulating lymphocytes with Ifn-?, anti-CD3, and interleukin-2 promotes the proliferation of a cell population coexpressing T-lymphocyte surface antigens such as CD3, CD8a, and CD25 as well as natural killer cell markers such as NK1.1, CD49, and CD69. These cells, referred to as cytokine-induced killer cells (CIKs), display cytotoxic activity against tumour cells, even without prior antigen presentation, and offer a new cell-based approach to the treatment of malignant diseases. Because CIKs are limited in vivo, strategies to optimize in vitro culture yield are required. In the last 10 years, mesenchymal stem cells (MSCs) have gathered considerable attention. Aside from their uses in tissue engineering and as support in haematopoietic stem cell transplantations, MSCs show notable immunomodulatory characteristics, providing further possibilities for therapeutic applications. In this study, we investigated the influence of murine MSCs on proliferation, phenotype, vitality, and cytotoxicity of murine CIKs in a coculture system. We found that CIKs in coculture proliferated within 7 days, with an average growth factor of 18.84, whereas controls grew with an average factor of 3.7 in the same period. Furthermore, higher vitality was noted in cocultured CIKs than in controls. Cell phenotype was unaffected by coculture with MSCs and, notably, coculture did not impact cytotoxicity against the tumour cells analysed. The findings suggest that cell–cell contact is primarily responsible for these effects. Humoral interactions play only a minor role. Furthermore, no phenotypical MSCs were detected after coculture for 4 h, suggesting the occurrence of immune reactions between CIKs and MSCs. Further investigations with DiD-labelled MSCs revealed that the observed disappearance of MSCs appears not to be due to differentiation processes.

Stolzing, Alexandra

2014-01-01

300

Stable long-term blood formation by stem cells in murine steady-state hematopoiesis.  

PubMed

Hematopoietic stem cells (HSCs) generate all mature blood cells during the whole lifespan of an individual. However, the clonal contribution of individual HSC and progenitor cells in steady-state hematopoiesis is poorly understood. To investigate the activity of HSCs under steady-state conditions, murine HSC and progenitor cells were genetically marked in vivo by integrating lentiviral vectors (LVs) encoding green fluorescent protein (GFP). Hematopoietic contribution of individual marked clones was monitored by determination of lentiviral integration sites using highly sensitive linear amplification-mediated-polymerase chain reaction. A remarkably stable small proportion of hematopoietic cells expressed GFP in LV-injected animals for up to 24 months, indicating stable marking of murine steady-state hematopoiesis. Analysis of the lentiviral integration sites revealed that multiple hematopoietic clones with both myeloid and lymphoid differentiation potential contributed to long-term hematopoiesis. In contrast to intrafemoral vector injection, intravenous administration of LV preferentially targeted short-lived progenitor cells. Myelosuppressive treatment of mice prior to LV-injection did not affect the marking efficiency. Our study represents the first continuous analysis of clonal behavior of genetically marked hematopoietic cells in an unmanipulated system, providing evidence that multiple clones are simultaneously active in murine steady-state hematopoiesis. PMID:22696148

Zavidij, Oksana; Ball, Claudia R; Herbst, Friederike; Oppel, Felix; Fessler, Sylvia; Schmidt, Manfred; von Kalle, Christof; Glimm, Hanno

2012-09-01

301

Murine coronavirus nonstructural protein ns2 is not essential for virus replication in transformed cells.  

PubMed Central

Two isolates of the murine hepatitis virus (MHV) strain JHM, which differed in their ability to express the nonstructural gene product ns2, were characterized. The MHV Wb3 isolate encodes a 30,000-molecular-weight ns2 protein that can be readily detected in infected cells by using a specific monoclonal antibody, MAb 2A. The MHV Wb1 isolate is a deletion mutant that lacks a functional ns2 gene and the transcriptional signals required for the synthesis of an ns2 mRNA. However, there are no obviously significant differences in the growth of the MHV Wb1 and MHV Wb3 isolates in continuous cell lines or in the synthesis of viral mRNAs or proteins in infected cells. These results demonstrate that the ns2 gene product is not essential for MHV replication in transformed murine cells and suggests that the function of the ns2 gene may only be manifest in vivo. Images

Schwarz, B; Routledge, E; Siddell, S G

1990-01-01

302

Immunostimulation of Murine Spleen Cells by Materials Associated with Bovine Milk Protein Fractions  

Microsoft Academic Search

Purified bovine milk proteins that were added to cultures of murine spleen cells significantly increased cell proliferation and production of immunoglobulin M. Casein and a whey mixture consisting of a- lactalbumin, bovine serum albumin, bovine gamma globulin, and b-lactoglobulin ( b-LG) were stimula- tory. Of the three b-LG preparations that are com- mercially available ( b-LG containing variants A and

K. F. Wong; N. Middleton; M. Montgomery; M. Dey; R. I. Carr

1998-01-01

303

Cleavage Map and Proteolytic Processing of the Murine Norovirus Nonstructural Polyprotein in Infected Cells  

Microsoft Academic Search

Murine norovirus (MNV) is presently the only member of the genus Norovirus in the Caliciviridae that can be propagated in cell culture. The goal of this study was to elucidate the proteolytic processing strategy of MNV during an authentic replication cycle in cells. A proteolytic cleavage map of the ORF1 polyprotein was generated, and the virus-encoded 3C-like (3CL) proteinase (Pro)

Stanislav V. Sosnovtsev; G. Belliot; K.-O. Chang; V. G. Prikhodko; L. B. Thackray; C. E. Wobus; S. M. Karst; H. W. Virgin; K. Y. Green

2006-01-01

304

Oligosaccharide modification by swainsonine treatment inhibits pulmonary colonization by B16-F10 murine melanoma cells  

Microsoft Academic Search

Oligosaccharide moieties of cell-surface glycoconjugates are thought to be involved in recognition events associated with tumor metastasis and invasion. Using swainsonine (SW), an inhibitor of Golgi ..cap alpha..-mannosidase II that results in the formation of hybrid-type oligosaccharides on N-linked glycoproteins, the authors have tested the hypothesis that specific glycan structures are required for pulmonary colonization by tumor cells. B16-F10 murine

M. J. Humphries; K. Matsumoto; S. L. White; K. Olden

1986-01-01

305

Methylcholanthrene-induced murine fibrosarcoma cell line BMT-11 secretes granulocyte colony-stimulating factor  

Microsoft Academic Search

Summary  Murine fibrosarcoma cell line BMT-11 was induced with 3-methyl-cholanthrene and maintained in culture. Transplantation of\\u000a BMT-11 into syngeneic C57 BL\\/6 mice produced leukocytosis consisting of marked increments of neutrophils and monocytes associated\\u000a with massive splenomegaly. In order to elucidate the mechanisms of this leukemoid reaction, we studied the changes occurring\\u000a in hematopoietic progenitor cells in BMT-11-transplanted mice. The numbers of

N. Ohhara; S. Okamura; S. Hayashi; T. Shibuya; F. Okada; M. Ishikawa; M. Hosokawa; H. Kobayashi; Y. Niho

1990-01-01

306

Role for TLR2 in NK Cell-Mediated Control of Murine Cytomegalovirus In Vivo  

Microsoft Academic Search

Natural killer (NK) cells are essential for the early control of murine cytomegalovirus (MCMV) infection. Here, we demonstrate that toll-like receptor 2 (TLR2) plays a role in the NK cell-mediated control of MCMV. TLR2 knockout (KO) mice had elevated levels of MCMV in the spleen and liver on day 4 postinfection compared to C57BL\\/6 mice. In vivo depletion of NK

Eva Szomolanyi-Tsuda; Xueya Liang; Raymond M. Welsh; Evelyn A. Kurt-Jones; Robert W. Finberg

2006-01-01

307

Blood-testis barrier dynamics are regulated by testosterone and cytokines via their differential effects on the kinetics of protein endocytosis and recycling in Sertoli cells  

Microsoft Academic Search

During spermatogenesis in the mamma- lian testis, preleptotene\\/leptotene spermatocytes differ- entiate from type B spermatogonia and traverse the blood-testis barrier (BTB) at stage VIII of the seminifer- ous epithelial cycle for further development. This timely movement of germ cells involves extensive junction re- structuring at the BTB. Previous studies have shown that these events are regulated by testosterone (T) and

Helen H. N. Yan; Dolores D. Mruk; Will M. Lee; C. Yan Cheng

2008-01-01

308

Murine and Human Myogenic Cells Identified by Elevated Aldehyde Dehydrogenase Activity: Implications for Muscle Regeneration and Repair  

Microsoft Academic Search

BackgroundDespite the initial promise of myoblast transfer therapy to restore dystrophin in Duchenne muscular dystrophy patients, clinical efficacy has been limited, primarily by poor cell survival post-transplantation. Murine muscle derived stem cells (MDSCs) isolated from slowly adhering cells (SACs) via the preplate technique, induce greater muscle regeneration than murine myoblasts, primarily due to improved post-transplantation survival, which is conferred by

Joseph B. Vella; Seth D. Thompson; Mark J. Bucsek; Minjung Song; Johnny Huard

2011-01-01

309

Expression and co-cytokine function of murine thioredoxin/adult T cell leukaemia-derived factor (ADF).  

PubMed

Human ADF (adult T cell leukaemia-derived factor), an isoform of thioredoxin, promotes proliferation of certain human lymphoid cell lines and is involved in many thiol-dependent reducing reactions. To study functional aspects of the murine homologue, we established inducible overexpression of murine ADF in E. coli and a purification method which led to an apparently homogeneous 14 kDa protein. This recombinant ADF was tested in proliferation assays with murine Th2 cells (D10.G4.1) and CTLL-2 cells. In synergy with IL-2, IL-4, IL-7 and IL-9 ADF displayed co-cytokine activity. These proliferative effects were neutralized by an affinity-purified polyclonal rabbit anti-ADF antiserum. The effects of ADF were critically dependent on the presence of 2-mercaptoethanol. Bacterial thioredoxin had similar effects on the proliferation of murine T cells. Thus, the thiol-related reducing capacity of these proteins is essential for their growth promoting activity. As investigated at the levels of mRNA and protein in several murine cell clones and lines as well as in mouse tissues ADF is expressed ubiquitously. Finally it could be demonstrated by competitive PCR that in contrast to cytokine mRNAs (e.g. IL-4 and IL-13) the expression of ADF mRNA in murine Th2 clones and spleen cells is not influenced by stimulation of these cells through the T cell receptor complex. Murine ADF therefore represents a protein constitutively expressed in a wide variety of cells with the capacity to enhance the proliferative effect of several cytokines on murine T cells. PMID:8742061

Blum, H; Röllinghoff, M; Gessner, A

1996-01-01

310

Purified murine granulocyte/macrophage progenitor cells express a high-affinity receptor for recombinant murine granulocyte/macrophage colony-stimulating factor  

SciTech Connect

Purified recombinant murine granulocyte/macrophage colony-stimulating factor (GM-CSF) was labeled with /sup 125/I and used to examine the GM-CSF receptor on unfractionated normal murine bone marrow cells, casein-induced peritoneal exudate cells, and highly purified murine granulocyte/macrophage progenitor cells (CFU-GM). CFU-GM were isolated from cyclophosphamide-treated mice by Ficoll-Hypaque density centrifugation followed by counterflow centrifugal elutriation. The resulting population had a cloning efficiency of 62-99% in cultures containing conditioned medium from pokeweed mitogen-stimulated spleen cells and 55-86% in the presence of a plateau concentration of purified recombinant murine GM-CSF. Equilibrium binding studies with /sup 125/I-labeled GM-CSF showed that normal bone marrow cells, casein-induced peritoneal exudate cells, and purified CFU-GM had a single class of high-affinity receptor. Affinity crosslinking studies demonstrated that /sup 125/I-labeled GM-CSF bound specifically to two species of M/sub r/ 180,000 and 70,000 on CFU-GM, normal bone marrow cells, and peritoneal exudate cells. The M/sub r/ 70,000 species is thought to be a proteolytic fragment of the intact M/sub r/ 180,000 receptor. The present studies indicate that the GM-CSF receptor expressed on CFU-GM and mature myeloid cells are structurally similar. In addition, the number of GM-CSF receptors on CFU-GM is twice the average number of receptors on casein-induced mature myeloid cells, suggesting that receptor number may decrease as CFU-GM mature.

Williams, D.E.; Bicknell, D.C.; Park, L.S.; Straneva, J.E.; Cooper, S.; Broxmeyer, H.E.

1988-01-01

311

Stimulation of cell proliferation in the subventricular zone by synthetic murine pheromones  

PubMed Central

Adult neurogenesis in female mice is known to be enhanced by exposure to soiled bedding from males, although the identity of the relevant chemosignals has remained unknown. Here we show that the previously recognized male murine pheromones, the farnesenes and 2-sec-butyl-4,5-dihydrothiazole (SBT), strongly increase cell proliferation in the subventricular zone (SVZ) of adult female mice, but not younger female mice. In addition, we found that a unique female murine pheromone, 2,5-dimethylpyrazine, facilitates similar changes in males. SBT stimulated cell proliferation in the SVZ of only adult females and not in young adult or pre- and post-puberty females. Our study suggests that pheromonal communication between males and females is enhancing reproductive success by controlling the estrous cycle and by promoting cell proliferation in a reciprocal manner.

Koyama, Sachiko; Soini, Helena A.; Foley, John; Novotny, Milos V.; Lai, Cary

2013-01-01

312

Murine and Human Myogenic Cells Identified by Elevated Aldehyde Dehydrogenase Activity: Implications for Muscle Regeneration and Repair  

PubMed Central

Background Despite the initial promise of myoblast transfer therapy to restore dystrophin in Duchenne muscular dystrophy patients, clinical efficacy has been limited, primarily by poor cell survival post-transplantation. Murine muscle derived stem cells (MDSCs) isolated from slowly adhering cells (SACs) via the preplate technique, induce greater muscle regeneration than murine myoblasts, primarily due to improved post-transplantation survival, which is conferred by their increased stress resistance capacity. Aldehyde dehydrogenase (ALDH) represents a family of enzymes with important morphogenic as well as oxidative damage mitigating roles and has been found to be a marker of stem cells in both normal and malignant tissue. In this study, we hypothesized that elevated ALDH levels could identify murine and human muscle derived cell (hMDC) progenitors, endowed with enhanced stress resistance and muscle regeneration capacity. Methodology/Principal Findings Skeletal muscle progenitors were isolated from murine and human skeletal muscle by a modified preplate technique and unfractionated enzymatic digestion, respectively. ALDHhi subpopulations isolated by fluorescence activate cell sorting demonstrated increased proliferation and myogenic differentiation capacities compared to their ALDHlo counterparts when cultivated in oxidative and inflammatory stress media conditions. This behavior correlated with increased intracellular levels of reduced glutathione and superoxide dismutase. ALDHhi murine myoblasts were observed to exhibit an increased muscle regenerative potential compared to ALDHlo myoblasts, undergo multipotent differentiation (osteogenic and chondrogenic), and were found predominately in the SAC fraction, characteristics that are also observed in murine MDSCs. Likewise, human ALDHhi hMDCs demonstrated superior muscle regenerative capacity compared to ALDHlo hMDCs. Conclusions The methodology of isolating myogenic cells on the basis of elevated ALDH activity yielded cells with increased stress resistance, a behavior that conferred increased regenerative capacity of dystrophic murine skeletal muscle. This result demonstrates the critical role of stress resistance in myogenic cell therapy as well as confirms the role of ALDH as a marker for rapid isolation of murine and human myogenic progenitors for cell therapy.

Vella, Joseph B.; Thompson, Seth D.; Bucsek, Mark J.; Song, Minjung; Huard, Johnny

2011-01-01

313

Differential gene expression in the murine gastric fundus lacking interstitial cells of Cajal  

PubMed Central

Background The muscle layers of murine gastric fundus have no interstitial cells of Cajal at the level of the myenteric plexus and only possess intramuscular interstitial cells and this tissue does not generate electric slow waves. The absence of intramuscular interstitial cells in W/WV mutants provides a unique opportunity to study the molecular changes that are associated with the loss of these intercalating cells. Method The gene expression profile of the gastric fundus of wild type and W/WV mice was assayed by murine microarray analysis displaying a total of 8734 elements. Queried genes from the microarray analysis were confirmed by semi-quantitative reverse transcription-polymerase chain reaction. Results Twenty-one genes were differentially expressed in wild type and W/WV mice. Eleven transcripts had 2.0–2.5 fold higher mRNA expression in W/WV gastric fundus when compared to wild type tissues. Ten transcripts had 2.1–3.9 fold lower expression in W/WV mutants in comparison with wild type animals. None of these genes have ever been implicated in any bowel motility function. Conclusions These data provides evidence that several important genes have significantly changed in the murine fundus of W/WV mutants that lack intramuscular interstitial cells of Cajal and have reduced enteric motor neurotransmission.

Daigo, Yataro; Takayama, Ichiro; Ponder, Bruce AJ; Caldas, Carlos; Ward, Sean M; Sanders, Kenton M; Fujino, Masayuki A

2003-01-01

314

Induction of Apoptosis and Inhibition of Cell Migration and Tube-Like Formation by Dihydroartemisinin in Murine Lymphatic Endothelial Cells  

Microsoft Academic Search

Dihydroartemisinin (DHA) is a semisynthesized agent from the artemisinin first extracted from the Chinese plant Artemisia annua. Previous studies have shown that artemisinin derivates, apart from their antimalarial activity, possess antitumor, antiangiogenic, and anti-inflammatory effects. In the present investigation, DHA was found to have a potent ability in influencing lymphatic endothelial cells (LECs) behavior. Murine LECs were isolated from benign

Jun Wang; Yan Guo; Bi-Cheng Zhang; Zheng-Tang Chen; Jian-Fei Gao

2007-01-01

315

Identification of replication competent murine gammaretroviruses in commonly used prostate cancer cell lines.  

PubMed

A newly discovered gammaretrovirus, termed XMRV, was recently reported to be present in the prostate cancer cell line CWR22Rv1. Using a combination of both immunohistochemistry with broadly-reactive murine leukemia virus (MLV) anti-sera and PCR, we determined if additional prostate cancer or other cell lines contain XMRV or MLV-related viruses. Our study included a total of 72 cell lines, which included 58 of the 60 human cancer cell lines used in anticancer drug screens and maintained at the NCI-Frederick (NCI-60). We have identified gammaretroviruses in two additional prostate cancer cell lines: LAPC4 and VCaP, and show that these viruses are replication competent. Viral genome sequencing identified the virus in LAPC4 and VCaP as nearly identical to another known xenotropic MLV, Bxv-1. We also identified a gammaretrovirus in the non-small-cell lung carcinoma cell line EKVX. Prostate cancer cell lines appear to have a propensity for infection with murine gammaretroviruses, and we propose that this may be in part due to cell line establishment by xenograft passage in immunocompromised mice. It is unclear if infection with these viruses is necessary for cell line establishment, or what confounding role they may play in experiments performed with these commonly used lines. Importantly, our results suggest a need for regular screening of cancer cell lines for retroviral "contamination", much like routine mycoplasma testing. PMID:21698104

Sfanos, Karen Sandell; Aloia, Amanda L; Hicks, Jessica L; Esopi, David M; Steranka, Jared P; Shao, Wei; Sanchez-Martinez, Silvia; Yegnasubramanian, Srinivasan; Burns, Kathleen H; Rein, Alan; De Marzo, Angelo M

2011-01-01

316

Identification of Replication Competent Murine Gammaretroviruses in Commonly Used Prostate Cancer Cell Lines  

PubMed Central

A newly discovered gammaretrovirus, termed XMRV, was recently reported to be present in the prostate cancer cell line CWR22Rv1. Using a combination of both immunohistochemistry with broadly-reactive murine leukemia virus (MLV) anti-sera and PCR, we determined if additional prostate cancer or other cell lines contain XMRV or MLV-related viruses. Our study included a total of 72 cell lines, which included 58 of the 60 human cancer cell lines used in anticancer drug screens and maintained at the NCI-Frederick (NCI-60). We have identified gammaretroviruses in two additional prostate cancer cell lines: LAPC4 and VCaP, and show that these viruses are replication competent. Viral genome sequencing identified the virus in LAPC4 and VCaP as nearly identical to another known xenotropic MLV, Bxv-1. We also identified a gammaretrovirus in the non-small-cell lung carcinoma cell line EKVX. Prostate cancer cell lines appear to have a propensity for infection with murine gammaretroviruses, and we propose that this may be in part due to cell line establishment by xenograft passage in immunocompromised mice. It is unclear if infection with these viruses is necessary for cell line establishment, or what confounding role they may play in experiments performed with these commonly used lines. Importantly, our results suggest a need for regular screening of cancer cell lines for retroviral “contamination”, much like routine mycoplasma testing.

Sfanos, Karen Sandell; Aloia, Amanda L.; Hicks, Jessica L.; Esopi, David M.; Steranka, Jared P.; Shao, Wei; Sanchez-Martinez, Silvia; Yegnasubramanian, Srinivasan; Burns, Kathleen H.; Rein, Alan; De Marzo, Angelo M.

2011-01-01

317

Peptidoglycan Induces Necrosis and Regulates Cytokine Production in Murine Trophoblast Stem Cells  

PubMed Central

Problem Intrauterine bacterial infection during pregnancy may lead to adverse outcome. The objective of this study was to assess whether peptidoglycan (PGN) derived from Gram-positive bacteria induces trophoblast stem (TS) cell death or alters TS cell cytokine production. Method of study TLR transcript expression was assessed by RT-PCR. Protein expression was determined by confocal microscopy or flow cytometry. 7-Aminoactinomycin D (7-AAD) staining was used to assess TS cell death. Morphological features of cell death were evaluated by transmission electron microscopy. The presence of cleaved caspase-3 and HMGB1 protein was examined by western blot. Cytokine levels in cell supernatants were determined using a mouse cytokine 23-plex panel. Results TLR2 and TLR4 protein was expressed from the 1-cell through the blastocyst stage of murine embryo development. Murine TS cells expressed TLR2 and TLR6 but not TLR1 or TLR4 RNA. Only TLR2 protein was detected at the plasma membrane of TS cells. PGN induced TS cell death by a caspase-3 independent mechanism. The cell death pathway induced by PGN was morphologically consistent with necrosis. Finally, PGN induced HMGB1 release and increased MIP-1? secretion while inhibiting the constitutive release of RANTES. Conclusion PGN-induced TS cell necrosis and the subsequent release of HMGB1 and MIP-1? may regulate an infection-induced inflammatory response at the maternal-fetal interface and thus may play a role in the pathogenesis of infection-associated pregnancy complications.

Rose, Jennifer A.; Rabenold, Jessica J.; Parast, Mana M.; Milstone, David S.; Abrahams, Vikki M.; Riley, Joan K.

2011-01-01

318

Single-nucleotide polymorphisms in the LRWD1 gene may be a genetic risk factor for Japanese patients with Sertoli cell-only syndrome.  

PubMed

Genetic mechanisms have been implicated as a cause of some cases of male infertility. Recently, ten novel genes involved in human spermatogenesis, including human LRWD1, have been identified by expression microarray analysis of human testictissue. The human LRWD1 protein mediates the origin recognition complex in chromatin, which is critical for the initiation of pre-replication complex assembly in G1 and chromatin organization in post-G1 cells. The Lrwd1 gene expression is specific to the testis in mice. Therefore, we hypothesized that mutation or polymorphisms of LRWD1 participate in male infertility, especially azoospermia. To investigate whether LRWD1 gene defects are associated with azoospermia caused by SCOS and meiotic arrest (MA), mutational analysis was performed in 100 and 30 Japanese patients by direct sequencing of the coding regions, respectively. Statistical analysis was performed for patients with SCOS and MA and in 100 healthy control men. No mutations were found in LRWD1; however, three coding single-nucleotide polymorphisms (SNP1-SNP3) could be detected in the patients. The genotype and allele frequencies in SNP1 and SNP2 were notably higher in the SCOS group than in the control group (P < 0.05). These results suggest the critical role of LRWD1 in human spermatogenesis. PMID:23445371

Miyamoto, T; Koh, E; Tsujimura, A; Miyagawa, Y; Saijo, Y; Namiki, M; Sengoku, K

2014-04-01

319

Intracerebral cell transplantation therapy for murine GM1 gangliosidosis  

Microsoft Academic Search

We performed a cell transplantation study to treat the brain involvement in lysosomal storage diseases. We used acid ?-galactosidase knock-out mice (BKO) from C57BL\\/6 as recipients. To minimize immune responses, we used cells derived from transgenic mice of C57BL\\/6 overexpressing the normal human ?-galactosidase. Fetal brain cells (FBC), bone marrow-derived mesenchymal stem cells (MSC), and mixed FBC and MSC cells

Tomo Sawada; Akemi Tanaka; Katsumi Higaki; Ayumi Takamura; Eiji Nanba; Toshiyuki Seto; Mitsuyo Maeda; Etsuko Yamaguchi; Junichiro Matsuda; Tunekazu Yamano

2009-01-01

320

Icariin promotes expression of PGC1?, PPAR?, and NRF-1 during cardiomyocyte differentiation of murine embryonic stem cells in vitro  

Microsoft Academic Search

Aim:To investigate the effect of icariin on the expression of peroxisome proliferator-activated receptor ? coactivator-1 alpha (PGC-1?), peroxisome proliferator-activated receptor alpha (PPAR?), and nuclear respiratory factor 1 (NRF-1) on cardiomyocyte differentiation of murine embryonic stem (ES) cells in vitro.Methods:The cardiomyocytes derived from murine ES cells were verified by immunocytochemistry using confocal laser scanning microscopy. Cardiac-specific sarcomeric proteins (ie ?-actinin, troponin

Ling Ding; Xing-guang Liang; Dan-yan Zhu; Yi-jia Lou

2007-01-01

321

Tellurite-induced oxidative stress leads to cell death of murine hepatocarcinoma cells.  

PubMed

Data regarding tellurium (Te) toxicity are scarce. Studies on its metabolism, performed mainly in bacteria, underline a major role of reactive oxygen species (ROS). We investigated whether tellurite undergoes redox cycling leading to ROS formation and cancer cell death. The murine hepatocarcinoma Transplantable Liver Tumor (TLT) cells were challenged with tellurite either in the presence or in the absence of different compounds as N-acetylcysteine (NAC), 3-methyladenine, BAPTA-AM, and catalase. NAC inhibition of tellurite-mediated toxicity suggested a major role of oxidative stress. Tellurite also decreased both glutathione (GSH) and ATP content by 57 and 80%, respectively. In the presence of NAC however, the levels of such markers were almost fully restored. Tellurite-mediated ROS generation was assessed both by using the fluorescent, oxidation-sensitive probe dichlorodihydrofluorescein diacetate (DCHF-DA) and electron spin resonance (ESR) spectroscopy to detect hydroxyl radical formation. Cell death occurs by a caspase-independent mechanism, as shown by the lack of caspase-3 activity and no cleavage of poly(ADP-ribose)polymerase (PARP). The presence of gamma-H2AX suggests tellurite-induced DNA strand breaking, NAC being unable to counteract it. Although the calcium chelator BAPTA-AM did show no effect, the rapid phosphorylation of eIF2alpha suggests that, in addition to oxidative stress, an endoplasmic reticulum (ER) stress may be involved in the mechanisms leading to cell death by tellurite. PMID:20213267

Sandoval, Juan M; Levêque, Philippe; Gallez, Bernard; Vásquez, Claudio C; Buc Calderon, Pedro

2010-08-01

322

Doxorubicin treatment induces tumor cell death followed by immunomodulation in a murine neuroblastoma model  

PubMed Central

Chemotherapy of malignant tumors induces tumor cell death. Numerous antitumor agents induce apoptosis of tumor cells, which are subsequently engulfed by phagocytes, initiating an immune reaction. The induction of immunogenic cell death by antitumor agents may be advantageous for antitumor immunity. The purpose of this study was to determine whether doxorubicin is capable of inducing an immunogenic reaction in murine neuroblastoma cells. The murine neuroblastoma cell line (neuro-2a cells) was cultured in a medium containing doxorubicin or cisplatin (CDDP), and induction of cell death was confirmed by cell viability assays. Cluster of differentiation (CD)8?+ lymphocytes were co-cultured with neuro-2a cells that had died following treatment with either doxorubicin or CDDP, and CD11b+ spleen cells or bone marrow-derived dendritic cells (BM-DCs) were added to the culture. Proliferation of CD8?+ lymphocytes and interferon (IFN)-? production were evaluated. When CD8?+ cells were co-cultured with doxorubicin-treated neuro-2a cells and BM-DCs, CD8?+ cells reacted to anti-CD3/CD28 antibody stimulation, proliferated and increased IFN-? production. IFN-? production was more effectively promoted by co-culture with doxorubicin-treated neuro-2a cells than by co-culture with CDDP-treated neuro-2a cells. These findings suggest that doxorubicin is capable of inducing immunogenic cell death in neuroblastoma cells, and thus has an immunological advantage for chemotherapy of neuroblastoma compared with CDDP. BM-DCs are considered to be the key antigen-presenting cells in the immune reaction following the induction of immunogenic neuroblastoma cell death and phagocytosis.

INOUE, SEIICHIRO; SETOYAMA, YUMIKO; ODAKA, AKIO

2014-01-01

323

Isolation, cultivation, and characterization of adult murine prostate stem cells  

PubMed Central

ABSTRACT/SUMMARY The successful isolation and cultivation of prostate stem cells will allow us to study their unique biological properties and their application in therapeutic approaches. Here we provide step-by-step procedures on the basis of previous work in our laboratory for: the harvesting of primary prostate cells from adolescent male mice by a modified enzymatic procedure; the isolation of an enriched population of prostate stem cells through cell sorting; the cultivation of prostate stem cells in vitro; and characterization of these cells and their stem-like activity, including in vivo tubule regeneration. Normally it will take approximately 8 hours to harvest prostate cells, isolate the stem cell enriched population, and set up the in vitro sphere assay. It will take up to 8 weeks to analyze the unique properties of the stem cells, including their regenerative capacity in vivo.

Lukacs, Rita U.; Goldstein, Andrew S.; Lawson, Devon A.; Cheng, Donghui; Witte, Owen N.

2010-01-01

324

The expression of TIPE1 in murine tissues and human cell lines.  

PubMed

Members of the tumor necrosis factor-alpha-induced protein-8 (TNFAIP8 or TIPE) family play important roles in immune homeostasis and cancer. TIPE1 (TNFAIP8-like 1) is a new member of the TIPE family that may regulate cell death. However, due to the lack of a suitable antibody, the nature of cells and tissues that express TIPE1 protein has not been determined. In this study, we generated a highly specific antibody to TIPE1 and examined TIPE1 expression in various murine tissues and human cell lines by immunohistochemistry, reverse transcription real-time PCR, and Western blot. We found that TIPE1 protein was detected in a wide variety of tissues in C57BL/6 mice, such as neurons in brain, hepatocytes, germ cells of female and male reproductive organs, muscular tissues, and a variety of cells of the epithelial origin, particularly those with secretory functions. TIPE1 protein was not expressed in mature T or B lymphocytes, but detectable in human B lymphoblast cell line HMy2.CIR and murine T cell line EL4. Furthermore, high levels of TIPE1 mRNA were detected in most human carcinoma cell lines, especially in cells transformed with viral genomes. These results indicate that TIPE1 may perform functions in cell secretion and carcinogenesis, but not in immunity. PMID:21600655

Cui, Jian; Zhang, Guizhong; Hao, Chunyan; Wang, Yan; Lou, Yunwei; Zhang, Wenqian; Wang, Juan; Liu, Suxia

2011-07-01

325

Pharmacological and molecular characterization of intrinsic and acquired doxorubicin resistance in murine tumor cell lines  

Microsoft Academic Search

We have studied the pharmacological parameters of doxorubicin resistance in three lines of murine cells selected by long-term culture in the presence of this drug or vincristine. A line originating from rat hepatoma spontaneously presented an intrinsic doxorubicin resistance as compared to the other lines, originating from a rat glioblastoma and from simian-virus-40-transformed mouse hepatocytes. This intrinsic resistance, as well

Brigitte Schott; Danielle Londos-Gagliardi; Colette Ries; Sylvie Huet; Jacques Robert

1993-01-01

326

Age-associated alterations in CXCL1 chemokine expression by murine B cells  

Microsoft Academic Search

BACKGROUND: The CXCL1 chemokines, macrophage inflammatory protein-2 (MIP-2) and cytokine-induced neutrophil chemoattractant (KC), have been shown to play a role in a number of pathophysiological disease states including endotoxin-induced inflammation and bacterial meningitis. While the expression of these chemokines has been identified in a variety of cell types in the mouse, little is known about their expression with murine B-lymphocytes.

Lina Hu; Vishwa Deep Dixit; Valeria de Mello-Coelho; Dennis D Taub

2004-01-01

327

Absence of a Specific Ganglioside Galactosyltransferase in Mouse Cells Transformed by Murine Sarcoma Virus  

PubMed Central

The ganglioside composition of a nonproducer subclone (derived from BALB/c 3T3 mouseembryo cells) transformed by Kirsten murine sarcoma virus was drastically altered compared to the nontransformed parental clone. The transformed clone was unable to synthesize the mono- and disialogangliosides, GM1 and GD1a, due to the complete absence of a specific galactosyltransferase. The lack of this enzyme activity was established by sensitive radiochemical and enzymological techniques in vivo and in vitro. Images

Fishman, Peter H.; Brady, Roscoe O.; Bradley, Roy M.; Aaronson, Stuart A.; Todaro, George J.

1974-01-01

328

Characterization of Progenitor Cells in Pulps of Murine Incisors  

Microsoft Academic Search

The continuous growth of rodent incisors requires the presence of stem cells capable of generating ameloblasts and odontoblasts. While epithelial stem cells giving rise to ameloblasts have been well-characterized, cells giving rise to the odontoblasts in incisors have not been fully characterized. The goal of this study was to gain insight into the potential population in dental pulps of unerupted

A. Balic; M. Mina

2010-01-01

329

In vivo developmental stages in murine natural killer cell maturation  

Microsoft Academic Search

Natural killer (NK) cells develop in the bone marrow, but their in vivo stages of maturation, expansion and acquisition of receptors that guide target cell specificity are not well defined. We describe here such stages of development. We also show that developing NK cells actively proliferate at a phenotypically distinguishable immature stage after they have acquired expression of Ly49 and

Sungjin Kim; Koho Iizuka; Hyun-Seok P. Kang; Ayotunde Dokun; Anthony R. French; Suellen Greco; Wayne M. Yokoyama

2002-01-01

330

Cytoplasmic superoxide dismutase and catalase activity and resistance to radiation lethality in murine tumor cells  

SciTech Connect

Reduced species of molecular oxygen are produced by the interaction of ionizing radiation with aqueous solutions containing molecular oxygen. The enzymes catalase and superoxide dismutase (SOD) are thought to function in vivo as scavengers of metabolically produced peroxide and superoxide respectively. SOD has been shown to protect against the lethal effects of ionizing radiation in vitro and in vivo. The authors have investigated the relationship between the cytosolic SOD catalase content and the sensitivity to radiation lethality of a number of murine cell lines (402AX, EL-4, MB-2T3, MB-4, MEL, P-815, SAI, SP-2, and SV-3T3). K/sub i/(CN/sup -/) for murine Cu-Zn-SOD was determined to be 6.8 x 10/sup -6/ M. No cytosolic Mn-SOD activity was found in any of the cell lines studied. No correlation was found between the cytosolic Cu-Zn-SOD or cytosolic catalase activity and the resistance to radiation lethality or the murine cell lines studied.

Davy, C.A.; Tesfay, Z.; Jones, J.; Rosenberg, R.C.; McCarthy, C.; Rosenberg, S.O.

1986-05-01

331

Relationships between DNA damage and the survival of murine bone marrow cells irradiated in situ  

SciTech Connect

The relationships between DNA damage and the survival of murine bone marrow cells irradiated in situ were examined. Cell survival was assayed by the ability of bone marrow cells from irradiated mice to form colonies in vitro (CFU-C). DNA double-strand breaks (DSBs) were measured by neutral (nondenaturing) filter elution and pulsed-field gel electrophoresis (PFGE). Double-strand breaks were measured in the proliferating bone marrow cells, identified by injecting the mice with [{sup 3}H]dThd at various times before {gamma} irradiation, as a model of the behavior of the radiosensitive target cells. To assess how the DNA lesions measured using these techniques correlated with cell killing, the effect of the radioprotective agent WR-2721 on the induction of DSBs in proliferating bone marrow cells was compared with its effect on CFU-C survival. WR-2721 protected against the killing of both granulocyte-macrophage and erythroid burst-forming CFU-C by a factor of about 2. In contrast, little (1.2-fold) protection was observed in the PFGE assay at radiation doses between 5 and 20 Gy. Similarly, at the lowest dose studied (5 Gy) there was little protection against DSBs as measured by neutral elution; only after doses of between 10 and 30 Gy was significant protection observed. Thus the previously reported predictive relationship between DSBs and cell survival in vitro does not appear to extend directly to murine bone marrow cells irradiated in vivo. 37 refs., 5 figs., 2 tabs.

Petrovecki, M.; Prager, A.; Terry, N.H.A.; Murray, D. [Univ. of Texas M.D. Anderson Cancer Center, Houston, TX (United States)

1994-06-01

332

CD133 is a marker for long-term repopulating murine epidermal stem cells.  

PubMed

Maintenance, repair, and renewal of the epidermis are thought to depend on a pool of dedicated epidermal stem cells (EpiSCs). Like for many somatic tissues, isolation of a nearly pure population of stem cells is a primary goal in cutaneous biology. We used a quantitative transplantation assay, using injection of keratinocytes into subcutis combined with limiting dilution analysis, to assess the long-term repopulating ability of putative murine EpiSC populations. Putative EpiSC populations were isolated by FACS sorting. The CD133(+) population and the subpopulation of CD133(+) cells that exhibits high mitochondrial membrane potential (D?m(hi)) were enriched for long-term repopulating EpiSCs versus unfractionated cells (3.9- and 5.2-fold, respectively). Evidence for self-renewal capacity was obtained by serial transplantation of long-term epidermal repopulating units derived from CD133(+) and CD133(+)??m(hi) keratinocytes. CD133(+) keratinocytes were multipotent and produced significantly more hair follicles than CD133(-) cells. CD133(+) cells were a subset of the previously described integrin ?6(+)CD34(+) bulge cell population, and 28.9±8.6% were label-retaining cells. Thus, murine keratinocytes within the CD133(+) and CD133(+)??m(hi) populations contain EpiSCs that regenerate the epidermis for the long term, are self-renewing, multipotent, and label-retaining cells. PMID:22763787

Charruyer, Alexandra; Strachan, Lauren R; Yue, Lili; Toth, Alexandra S; Cecchini, Gary; Mancianti, Maria L; Ghadially, Ruby

2012-11-01

333

No evidence for ? cell neogenesis in murine adult pancreas.  

PubMed

Whether facultative ? cell progenitors exist in the adult pancreas is a major unsolved question. To date, lineage-tracing studies have provided conflicting results. To track ? cell neogenesis in vivo, we generated transgenic mice that transiently coexpress mTomato and GFP in a time-sensitive, nonconditional Cre-mediated manner, so that insulin-producing cells express GFP under control of the insulin promoter, while all other cells express mTomato (INSCremTmG mice). Newly differentiated ? cells were detected by flow cytometry and fluorescence microscopy, taking advantage of their transient coexpression of GFP and mTomato fluorescent proteins. We found that ? cell neogenesis predominantly occurs during embryogenesis, decreases dramatically shortly after birth, and is completely absent in adults across various models of ? cell loss, ? cell growth and regeneration, and inflammation. Moreover, we demonstrated upregulation of neurogenin 3 (NGN3) in both proliferating ducts and preexisting ? cells in the ligated pancreatic tail after pancreatic ductal ligation. These results are consistent with some recent reports, but argue against the widely held belief that NGN3 marks cells undergoing endocrine neogenesis in the pancreas. Our data suggest that ? cell neogenesis in the adult pancreas occurs rarely, if ever, under either normal or pathological conditions. PMID:23619362

Xiao, Xiangwei; Chen, Zean; Shiota, Chiyo; Prasadan, Krishna; Guo, Ping; El-Gohary, Yousef; Paredes, Jose; Welsh, Carey; Wiersch, John; Gittes, George K

2013-05-01

334

FAAP, a novel murine protein, is involved in cell adhesion through regulating vinculin-paxillin association.  

PubMed

Focal adhesion associated protein (FAAP), encoded by murine D10Wsu52e gene, is highly homologous to human HSPC117, which interacts with vinculin and talin. HeLa cells transfected with FAAP exhibited normal adhesion incorporation but showed impaired cell spreading, and restrained focal adhesion translocation. Moreover, FAAP facilitated vinculin-paxillin association, decreased interaction of paxillin-focal adhesion kinase and inhibited the phosphorylation of extracellular signal-regulated kinase. Together, these results suggest that FAAP, by virtue of modulating interaction of adhesion molecules, regulates cell adhesion dynamics. PMID:18508721

Hu, Jinsong; Teng, Junlin; Ding, Naizheng; He, Mei; Sun, Yuhui; Yu, Albert Cheung Hoi; Chen, Jianguo

2008-01-01

335

Macrophage activation factor from EL-4, a murine T-cell line: antigenic characterization by hamster monoclonal antibodies to murine interferon-gamma.  

PubMed

A cloned variant of the EL-4 murine T-cell line treated with phorbol myristate acetate (PMA) releases a factor that activates macrophages for nonspecific tumor cytotoxicity. This macrophage activation factor (MAF) is both physicochemically (Mr 25,000; pH 2 stable) and biologically different from interferon-gamma (IFN-gamma). However, EL-4 MAF may represent a breakdown product or otherwise altered fragment of IFN-gamma. We examined this possibility with a unique pair of hamster monoclonal antibodies against different epitopes of murine IFN-gamma. Both antibodies inhibited IFN-gamma-induced fibroblast antiviral activity; H21 but not H1 antibody also inhibited lymphokine (LK)-induced macrophage-mediated tumor cytotoxicity. Neither antibody, however, had any effect on the EL-4 MAF throughout a broad dose response. Moreover, passage through a H21 immunoaffinity chromatography column or addition of staphylococcal protein A and antibody completely inhibited LK-induced macrophage tumoricidal activity but did not affect the activity in EL-4 MAF. Identical effects in both fluid and solid phase were observed with polyclonal rabbit antisera to murine IFN-gamma. Results with all of these antibodies strongly suggest that the EL-4 MAF and murine IFN-gamma are antigenically distinct. PMID:3109744

Meltzer, M S; Gilbreath, M J; Crawford, R M; Schreiber, R D; Nacy, C A

1987-07-01

336

Natural species-restricted attachment of human and murine T lymphocytes to various cells.  

PubMed Central

Murine and human T lymphocytes bear on their surface a receptor that confers on them the ability to attach to a variety of target cells from the same species, derived in vivo and in vitro. Thymocytes and activated T cells attached readily to target cells, while blood T lymphocytes were able to do so only after the removal of sialic acid from either their cell membrane or that of the target cell. The natural attachment (NA) receptor and the corresponding site on the target cells are trypsin sensitive and the conjugation between them is temperature dependent. The phenomenon may be a manifestation of self recognition in a broader sense--recognizing the species--which is also reflected in the reactivity of mitogen-activated T cells and specific immune responses against allo- or other antigens expressed on target cell surfaces. Images

Galili, U; Galili, N; Vanky, F; Klein, E

1978-01-01

337

Aryl hydrocarbon receptor controls murine mast cell homeostasis.  

PubMed

We propose that the aryl hydrocarbon receptor (AhR), a unique chemical sensor, is critical in controlling mast cell differentiation, growth, and function in vitro and in vivo. In antigen-stimulated mast cells, exposure to AhR ligands resulted in a calcium- and reactive oxygen species (ROS)-dependent increase of reversible oxidation in and reduced activity of SHP-2 phosphatase, leading to enhanced mast cell signaling, degranulation, and mediator and cytokine release, as well as the in vivo anaphylactic response. Surprisingly, significant mast cell deficiency was noted in AhR-null mice due to defective calcium signaling and mitochondrial function, concomitant with reduced expression of c-kit and cytosolic STAT proteins, as well as enhanced intracellular ROS and apoptosis. Consequently, AhR-null mast cells responded poorly to stimulation, demonstrating a critical role of AhR signaling in maintaining mast cell homeostasis. PMID:23462117

Zhou, Yufeng; Tung, Hui-Ying; Tsai, Ying-Ming; Hsu, Shih-Chang; Chang, Hui-Wen; Kawasaki, Hirokazu; Tseng, Hsiao-Chun; Plunkett, Beverly; Gao, Peisong; Hung, Chih-Hsing; Vonakis, Becky M; Huang, Shau-Ku

2013-04-18

338

Aryl hydrocarbon receptor controls murine mast cell homeostasis  

PubMed Central

We propose that the aryl hydrocarbon receptor (AhR), a unique chemical sensor, is critical in controlling mast cell differentiation, growth, and function in vitro and in vivo. In antigen-stimulated mast cells, exposure to AhR ligands resulted in a calcium- and reactive oxygen species (ROS)-dependent increase of reversible oxidation in and reduced activity of SHP-2 phosphatase, leading to enhanced mast cell signaling, degranulation, and mediator and cytokine release, as well as the in vivo anaphylactic response. Surprisingly, significant mast cell deficiency was noted in AhR-null mice due to defective calcium signaling and mitochondrial function, concomitant with reduced expression of c-kit and cytosolic STAT proteins, as well as enhanced intracellular ROS and apoptosis. Consequently, AhR-null mast cells responded poorly to stimulation, demonstrating a critical role of AhR signaling in maintaining mast cell homeostasis.

Zhou, Yufeng; Tung, Hui-Ying; Tsai, Ying-Ming; Hsu, Shih-Chang; Chang, Hui-Wen; Kawasaki, Hirokazu; Tseng, Hsiao-Chun; Plunkett, Beverly; Gao, Peisong; Hung, Chih-Hsing; Vonakis, Becky M.

2013-01-01

339

Murine bone marrow cell line producing colony-stimulating factor  

SciTech Connect

A cell line (H-1) derived from the adherent layer of a 14-wk-old Dexter bone marrow culture has been maintained as cloned and uncloned lines through 21 passages at the time of these studies. These cell lines develop many fat droplets as they age and become confluent. The uncloned line produces increasing amounts of colony-stimulating activity as the cells become confluent. Feeder-layers or supernatants from the nonconfluent or confluent fat-laden cells stimulate the formation of greater numbers of colonies derived from cultures of colony-forming units (CFU) than does medium from L cell culture containing colony-stimulating factor (CSF). Antibody to the CSF-containing medium from L cell culture neutralizes the colony-stimulating activity, thus showing immunologic similarity to a known molecular species that stimulates colony production in a CFU culture that produces granulocyte or macrophage populations, or both.

Harigaya, K. (Brookhaven National Lab., Upton, NY); Cronkite, E.P.; Miller, M.E.; Shadduck, R.K.

1981-11-01

340

Characterization of Murine Lung Dendritic Cells Infected with Mycobacterium tuberculosis  

Microsoft Academic Search

phage colony-stimulating factor, 99% of cells were CD11c1 and had a morphology typical of immature dendritic cells. These cells were negative for CD34, CD14, and CD8a antigens but expressed low levels of the myeloid marker F4\\/80 and moderate levels of MAC3. All expressed high levels of CD11a (LFA-1), CD11b (Mac1), and CD54 antigens, with low levels of class II major

MERCEDES GONZALEZ-JUARRERO; IAN M. ORME

2001-01-01

341

Extracortical origin of some murine subplate cell populations.  

PubMed

The subplate layer, the deepest cortical layer in mammals, has important roles in cerebral cortical development. The subplate contains heterogeneous cell populations that are morphologically diverse, with several projection targets. It is currently assumed that these cells are generated in the germinative zone of the earliest cortical neuroepithelium. Here we identify a pallial but extracortical area located in the rostromedial telencephalic wall (RMTW) that gives rise to several cell populations. Postmitotic neurons migrate tangentially from the RMTW toward the cerebral cortex. Most RMTW-derived cells are incorporated into the subplate layer throughout its rostrocaudal extension, with others contributing to the GABAergic interneuron pool of cortical layers V and VI. PMID:24778253

Pedraza, María; Hoerder-Suabedissen, Anna; Albert-Maestro, María Amparo; Molnár, Zoltán; De Carlos, Juan A

2014-06-10

342

Regulation of murine hematopoietic stem cell quiescence by Dmtf1  

PubMed Central

The cell-cycle status of hematopoietic stem cells (HSCs) is tightly regulated, most likely to balance maintenance of stem-cell status through quiescence and expansion/differentiation of the hematopoietic system. Tumor-suppressor genes (TSGs), with their cell cycle–regulatory functions, play important roles in HSC regulation. The cyclin-D binding myb-like transcription factor 1 (Dmtf1) was recently recognized as a TSG involved in human cancers by repressing oncogenic Ras/Raf signaling. However, the role of Dmtf1 in the hematopoietic system is entirely unknown. In the present study, we demonstrate that Dmtf1 regulates HSC function under both steady-state and stress conditions. Dmtf1?/? mice showed increased blood cell counts in multiple parameters, and their progenitor cells had increased proliferation and accelerated cell-cycle progression. In addition, long-term HSCs from Dmtf1?/? mice had a higher self-renewal capacity that was clearly demonstrated in secondary recipients in serial transplantation studies. Dmtf1?/? BM cells showed hyper proliferation after 5-fluorouracil–induced myeloablation. Steady-state expression and Induction of CDKN1a (p21) and Arf were impaired in HSCs from Dmtf1?/? mice. The function of Dmtf1 was mediated by both Arf-dependent and Arf-independent pathways. Our results implicate Dmtf1 in the regulation of HSC function through novel cell cycle–regulatory mechanisms.

Kobayashi, Michihiro

2011-01-01

343

Accumulation of invariant NKT cells into inflamed skin in a novel murine model of nickel allergy.  

PubMed

Nickel (Ni) can cause delayed-type hypersensitivity reactions, which are thought to be mediated by the accumulation of T cells into inflamed skin. Accumulated T cells at the developmental stages in metal allergy are poorly characterized because a suitable animal model has not been established. To investigate the accumulated T cells in allergic inflamed skin, we generated a novel murine model of Ni-induced allergy. The murine model of Ni allergy was induced by two sensitizations of Ni plus lipopolysaccharide solution into the groin followed by three challenges with Ni solution into the footpad. Here we show that a specific TCR repertoire bearing V?14J?18, called natural killer (NK) T cells, was expanded monoclonally in BALB/c or C57BL/6 mice. Accumulation of NKT cells was characterized as CD4(+) or CD4(-)CD8(-) T cells. These results suggested that NKT cells are major pathogenic T cells at the elicitation phase of Ni allergy. PMID:23978680

Eguchi, Takanori; Kumagai, Kenichi; Kobayashi, Hiroshi; Shigematsu, Hiroaki; Kitaura, Kazutaka; Suzuki, Satsuki; Horikawa, Tatsuya; Hamada, Yoshiki; Ogasawara, Kouetsu; Suzuki, Ryuji

2013-01-01

344

A murine model of hepatic veno-occlusive disease induced by allogeneic hematopoietic stem cell transplantation.  

PubMed

Hepatic veno-occlusive disease (HVOD) is a life-threatening complication of bone marrow stem cell transplantation. The understanding of this clinical condition is hampered by the lack of suitable animal models. Here, we present a murine (BALB/c-based) model of HVOD induced by allogeneic hematopoietic stem cell transplantation (allo-HSCT). The chimerism rate of bone marrow was measured on days 5 and 10, while the chimerism rate of peripheral blood was measured on day 15 after allo-HSCT. Percentages of peripheral reticulocytes and serum levels of bilirubin and alanine aminotransferase (as liver function tests) were measured on days 5, 10, 15, 20, and 30. Livers were obtained on days 5, 10, 15, 20, and 30, and fixed in formaldehyde or glutaric dialdehyde. Liver slices were processed using the hematoxylin-eosin, Masson's trichrome, or immunohistochemistry staining, and examined by light or transmission electron microscopy. Sinusoidal damages were the earliest pathological changes occurring in the allo-HSCT-induced HVOD, followed by coagulative necrosis of liver cells. The liver cell necrosis was later attenuated and sinusoidal endothelial cell morphology improved. However, on day 30, the edema and necrosis of liver cells became aggravated again. Furthermore, sinusoidal lining cell regeneration and partly attenuated liver cell necrosis were followed by the moderate to severe central vein fibrosis. In conclusion, we have successfully established a murine model of HSCT-HVOD. This model develops moderate to severe HVOD which cannot heal without intervention. PMID:23579582

Zeng, Lingyu; An, Licai; Fang, Ting; Pan, Bin; Sun, Haiying; Chen, Chong; Cao, Jiang; Li, Zhenyu; Xu, Kailin

2013-12-01

345

Cultured human bone marrow-derived mast cells, their similarities to cultured murine E-mast cells.  

PubMed Central

Homogeneous populations of human mast cells were differentiated and grown by culturing bone marrow cells in the presence of conditioned medium derived from lectin-stimulated human peripheral blood mononuclear cells. The cells obtained were similar in ultrastructure, proteoglycan type and lipid products generated upon calcium ionophore A23187, and immunological activation to the murine E-mast cells (E-MC) differentiated in culture containing IL-3. Fluorescence analysis revealed that the human E-MC expressed IgE-Fc receptors which retained bound IgE through several washes. These cells did not express cell-surface lymphoid determinants (T11, T4, T8 and B4) and myeloid determinants 'My'. However, 40% of these cells expressed monocytic surface determinants, such as M-1. The amount of histamine that was found per 10(6) cells was 525 +/- 106 ng (mean +/- SE, n = 4). These cultured mast cells possessed granular chondroitin sulphate E proteoglycan of about 180,000 MW. Following activation with either calcium ionophore A23187 or anti-hIgE challenge, these mast cells released their preformed mediators and generated mainly leukotriene C4 leukotriene B4, and platelet-activating factor. In conclusion, according to all of these criteria, these human cultured mast cells show many similarities to the murine cultured E-mast cells, and therefore could be considered as the culture analogue of the human intestinal E-mast cells identified recently. Images Figure 1

Gilead, L; Rahamim, E; Ziv, I; Or, R; Razin, E

1988-01-01

346

H4 Histamine Receptors Mediate Cell Cycle Arrest in Growth Factor-Induced Murine and Human Hematopoietic Progenitor Cells  

PubMed Central

The most recently characterized H4 histamine receptor (H4R) is expressed preferentially in the bone marrow, raising the question of its role during hematopoiesis. Here we show that both murine and human progenitor cell populations express this receptor subtype on transcriptional and protein levels and respond to its agonists by reduced growth factor-induced cell cycle progression that leads to decreased myeloid, erythroid and lymphoid colony formation. H4R activation prevents the induction of cell cycle genes through a cAMP/PKA-dependent pathway that is not associated with apoptosis. It is mediated specifically through H4R signaling since gene silencing or treatment with selective antagonists restores normal cell cycle progression. The arrest of growth factor-induced G1/S transition protects murine and human progenitor cells from the toxicity of the cell cycle-dependent anticancer drug Ara-C in vitro and reduces aplasia in a murine model of chemotherapy. This first evidence for functional H4R expression in hematopoietic progenitors opens new therapeutic perspectives for alleviating hematotoxic side effects of antineoplastic drugs.

Petit-Bertron, Anne-France; Machavoine, Francois; Defresne, Marie Paule; Gillard, Michel; Chatelain, Pierre; Mistry, Prakash

2009-01-01

347

Gene expression profiling of murine T-cell lymphoblastic lymphoma identifies deregulation of S-phase initiating genes?  

PubMed Central

In a search for genes and pathways implicated in T-cell lymphoblastic lymphoma (T-LBL) development, we used a murine lymphoma model, where mice of the NMRI-inbred strain were inoculated with murine leukemia virus mutants. The resulting tumors were analyzed by integration analysis and global gene expression profiling to determine the effect of the retroviral integrations on the nearby genes, and the deregulated pathways in the tumors. Gene expression profiling identified increased expression of genes involved in the minichromosome maintenance and origin of recognition pathway as well as downregulation in negative regulators of G1/S transition, indicating increased S-phase initiation in murine T-LBLs.

Dabrowska, Magdalena Julia; Ejegod, Ditte; Lassen, Louise Berkhoudt; Johnsen, Hans Erik; Wabl, Matthias; Pedersen, Finn Skou; Dybkaer, Karen

2014-01-01

348

Efficient propagation of betanodavirus in a murine astrocytoma cell line  

Microsoft Academic Search

Betanodavirus, a bipartite RNA virus of fishes and a member of Nodaviridae family, targets nervous tissues and is the causative agent of viral nervous necrosis in marine farmed fish. Betanodavirus is thought to be propagated only in fish cells because betanodavirus has only been isolated in fish and it is not well propagated in mammalian culture cells. However, the host

Naoki Takizawa; Kei Adachi; Tohru Ichinose; Nobuyuki Kobayashi

2008-01-01

349

Augmentation of Murine Natural Killer Cell Activity by Manganese Chloride.  

National Technical Information Service (NTIS)

Natural Killer (NK) cell activity of spleen cells from male CBA/J mice was augmented by a single parenteral injection of MnCl2 administered 1 day prior to testing by in vitro and in vivo isotope release assays. Increased cytotoxic activity was observed in...

R. Rogers R. Garner M. Riddle R. Luebke R. Smialowicz

1983-01-01

350

The effects of simulated hypogravity on murine bone marrow cells  

NASA Technical Reports Server (NTRS)

Mouse bone marrow cells grown in complete medium at unit gravity were compared with a similar population cultured in conditions that mimic some aspects of microgravity. After the cells adjusted to the conditions that simulated microgravity, they proliferated as fetal or oncogenic populations; their numbers doubled in twelve hour periods. Differentiated subpopulations were depleted from the heterogeneous mixture with time and the undifferentiated hematopoietic stem cells increased in numbers. The cells in the control groups in unit gravity and those in the bioreactors in conditions of microgravity were monitored under a number of parameters. Each were phenotyped as to cell surface antigens using a panel of monoclonal antibodies and flow cytometry. Other parameters compared included: pH, glucose uptake, oxygen consumption and carbon-dioxide production. Nuclear DNA was monitored by flow cytometry. Functional responses were studied by mitogenic stimulation by various lectins. The importance of these findings should have relevance to the space program. Cells should behave predictably in zero gravity; specific populations can be eliminated from diverse populations and other populations isolated. The availability of stem cell populations will enhance both bone marrow and gene transplant programs. Stem cells will permit developmental biologists study the paths of hematopoiesis.

Lawless, Desales

1989-01-01

351

AN IN VITRO MODEL FOR MURINE URETERIC EPITHELIAL CELLS  

EPA Science Inventory

This report presents a model developed to study growth and differentiation of primary cultures of ureteric epithelial cells from embryonic C57BL/6N mouse urinary tracts. Single cells were resuspended in medium and plated onto transwells coated with collagen IV and laminin. Basa...

352

Intracerebral cell transplantation therapy for murine GM1 gangliosidosis.  

PubMed

We performed a cell transplantation study to treat the brain involvement in lysosomal storage diseases. We used acid beta-galactosidase knock-out mice (BKO) from C57BL/6 as recipients. To minimize immune responses, we used cells derived from transgenic mice of C57BL/6 overexpressing the normal human beta-galactosidase. Fetal brain cells (FBC), bone marrow-derived mesenchymal stem cells (MSC), and mixed FBC and MSC cells were prepared and injected into the ventricle of newborn BKO mouse brain. The mice were examined at 1, 2, 4, and 8 weeks and 6 months after injection. In each experiment, the injected cells migrated into the whole brain effectively and survived for at least 8 weeks. Decrease in ganglioside GM1 level was also observed. FBC could survive for 6 months in recipient brain. However, the number of transplanted FBC decreased. In the brains of MSC- or mixed cell-treated mice, no grafted cells could be found at 6 months. To achieve sufficient long-term effects on the brain, a method of steering the immune response away from cytotoxic responses or of inducing tolerance to the products of therapeutic genes must be developed. PMID:19118961

Sawada, Tomo; Tanaka, Akemi; Higaki, Katsumi; Takamura, Ayumi; Nanba, Eiji; Seto, Toshiyuki; Maeda, Mitsuyo; Yamaguchi, Etsuko; Matsuda, Junichiro; Yamano, Tunekazu

2009-11-01

353

Murine model of leptomeningeal dissemination using human medulloblastoma cells.  

PubMed

An experimental model of meningeal dissemination was developed by intracisternal inoculation of human medulloblastoma (ONS-76) cells into nude mice. All mice died within 65 days after inoculation of 1 x 10(7) tumor cells. The median survival time was 56 days. Clinical signs and histological findings were similar to those in medulloblastoma patients with meningeal dissemination. Immunohistochemical studies showed that ONS-76 cells in the subarachnoid space expressed major histocompatibility complex (MHC) class I antigens until 20 days after inoculation. After 30 days, expression of MHC class I antigens decreased and cells began to proliferate rapidly. Expression of MHC class I antigens on tumor cells may result in effective recognition by the host immune system. PMID:1726223

Yamada, M; Shimizu, K; Tamura, K; Moriuchi, S; Mabuchi, E; Park, K C; Miyao, Y; Hayakawa, T

1991-12-01

354

Putative oncogenic role of the erythropoietin receptor in murine and human erythroleukemia cells.  

PubMed

To determine whether the erythropoietin receptor (Epo-R) plays a role in the course of malignant erythropoietic disorders, this gene was studied in murine and human erythroleukemia cells. An altered Epo-R gene was found in a murine Friend erythroleukemia cell line, FCL1, due to a spleen focus-forming virus (SFFV) long terminal repeat insertion within the noncoding region of the first exon, leading to Epo-R mRNA overexpression. A similar mechanism of Epo-R activation has previously been described in the T3CL-2 Friend erythroleukemia cell line. An elevated number of Epo-binding sites has been observed in two human erythroleukemia cell lines, TF-1 and UT7. In UT7 cells, homogeneously staining region of the short arm of chromosome 19 [hsr (19)] was evidenced, which contained an amplification of the Epo-R gene. This Epo-R gene amplification was confirmed by the quantification of Southern blots in which the intensity of the Epo-R signal was compared in UT7 DNA and in DNA from normal cells. The Epo-R gene was present in UT7 at a mean number of seven to eight copies per cell. Interestingly, the Epo-R gene was rearranged; the breakpoint region was located near the 3' end of the gene, 3 kb downstream from the end of the last exon. Taken together, these results suggest that, in both murine and human systems, genetic alterations of the Epo-R gene are not rare events and could be involved in the occurrence of the erythroleukemic process. PMID:8142650

Chretien, S; Moreau-Gachelin, F; Apiou, F; Courtois, G; Mayeux, P; Dutrillaux, B; Cartron, J P; Gisselbrecht, S; Lacombe, C

1994-04-01

355

Murine mast cells secrete and respond to interleukin-33.  

PubMed

Interleukin-33 (IL-33) appears to play a crucial role in the expression of allergic diseases, but its cellular source and regulatory mechanisms remain to be fully elucidated. Mast cells, one of the major effecter cell populations in mediating allergy, express high levels of IL-33 receptor, ST2, and have been shown to express IL-33 transcripts. In this study, we aimed to examine the secretion of IL-33 in mast cells and their response to IL-33. We have successfully detected secreted IL-33 from cell supernatants through a modified enzyme-linked immunosorbent assay (ELISA) technique-cell-based ELISA. Activation of bone marrow-derived cultured mast cells (BMMCs) by crosslinkage of an antigen [ovalbumin (OVA)] and OVA-specific IgE mAbs significantly induced the expression of IL-33 transcripts, cytosolic and secreted proteins. In addition, the Toll-like receptor (TLR) 2 and TLR-9 ligands could trigger IL-33 mRNA expression. Exposure of BMMCs to IL-33 significantly increased the levels of IL-13 and IL-6 expression, concomitant with enhanced activation of mitogen-activated protein kinase (MAPKs) (ERK, p38, and JNK) and nuclear factor-kappa B. These results suggest that mouse BMMCs are capable of producing and serving as endogenous sources of IL-33, and that IL-33 plays an important role in regulating mast cell functions. PMID:24028396

Tung, Hui-Ying; Plunkett, Beverly; Huang, Shau-Ku; Zhou, Yufeng

2014-03-01

356

Identification of molecular markers of bipolar cells in the murine retina  

PubMed Central

Retinal bipolar neurons serve as relay interneurons that connect rod and cone photoreceptor cells to amacrine and ganglion cells. They exhibit diverse morphologies essential for correct routing of photoreceptor cell signals to specific postsynaptic amacrine and ganglion cells. The development and physiology of these interneurons have not been completely defined molecularly. Despite previous identification of genes expressed in several bipolar cell subtypes, molecules that mark each bipolar cell type still await discovery. In this report, novel genetic markers of murine bipolar cells were found. Candidates were initially generated by using microarray analysis of single bipolar cells and mining of retinal serial analysis of gene expression (SAGE) data. These candidates were subsequently tested for expression in bipolar cells by RNA in situ hybridization. Ten new molecular markers were identified, five of which are highly enriched in their expression in bipolar cells within the adult retina. Double-labeling experiments using probes for previously characterized subsets of bipolar cells were performed to identify the subtypes of bipolar cells that express the novel markers. Additionally, the expression of bipolar cell genes was analyzed in Bhlhb4 knockout retinas, in which rod bipolar cells degenerate postnatally, to delineate further the identity of bipolar cells in which novel markers are found. From the analysis of Bhlhb4 mutant retinas, cone bipolar cell gene expression appears to be relatively unaffected by the degeneration of rod bipolar cells. Identification of molecular markers for the various subtypes of bipolar cells will lead to greater insights into the development and function of these diverse interneurons.

Kim, Douglas S; Ross, Sarah E; Trimarchi, Jeffrey M; Aach, John; Greenberg, Michael E; Cepko, Constance L

2008-01-01

357

The surface antigen CD45R identifies a population of estrogen-regulated murine marrow cells that contain osteoclast precursors.  

PubMed

We examined the osteoclastogenic potential of murine bone marrow cells that were fractionated according to their expression of the surface antigen CD45R. Osteoclast-like cells (OCL) with many authentic osteoclast characteristics readily formed in purified CD45R(+) murine bone marrow cell cultures after treatment with receptor activator of nuclear factor kappaB ligand (RANKL) and M-CSF. Ovariectomy (Ovx) caused a 1.5- to 2-fold increase in OCL number in unfractionated and CD45R(+) murine bone marrow cell cultures without affecting OCL formation in CD45R(-) marrow cells. Limiting dilution assays confirmed that Ovx caused an increase in osteoclast precursor cell number in CD45R(+) but not CD45R(-) cells. Mice deficient in the type 1 IL-1 receptor (IL-1R1 KO) do not lose bone mass after Ovx. We found that unfractionated, CD45R(+), and CD45R(-) bone marrow cells from IL-1R1 KO mice showed no increase in OCL formation in vitro after Ovx. In both the wild-type (WT) and the IL-1R1 KO mice Ovx was associated with a 2-fold increase in pre-B-lymphocytes. About 1.3-3.5% of murine marrow cells expressed surface RANK (the receptor for RANKL) while about 11.9-15% of murine bone marrow cells expressed c-Fms (the receptor for M-CSF). There was little effect of Ovx on cells expressing either RANK or c-Fms. These results demonstrate that CD45R expression identifies a subset of murine bone marrow cells whose ability to form OCL in vivo is regulated by estrogen in WT but not IL-1R1 KO cells. The effects of estrogen on bone mass may be related to these responses. PMID:12810165

Katavi?, Vedran; Grcevi?, Danka; Lee, Sun Kyeong; Kalinowski, Judith; Jastrzebski, Sandra; Dougall, William; Anderson, Dirk; Puddington, Lynn; Aguila, H Leonardo; Lorenzo, Joseph A

2003-06-01

358

Polycomb complexes repress developmental regulators in murine embryonic stem cells  

Microsoft Academic Search

The mechanisms by which embryonic stem (ES) cells self-renew while maintaining the ability to differentiate into virtually all adult cell types are not well understood. Polycomb group (PcG) proteins are transcriptional repressors that help to maintain cellular identity during metazoan development by epigenetic modification of chromatin structure. PcG proteins have essential roles in early embryonic development and have been implicated

Laurie A. Boyer; Kathrin Plath; Julia Zeitlinger; Tobias Brambrink; Lea A. Medeiros; Tong Ihn Lee; Stuart S. Levine; Marius Wernig; Adriana Tajonar; Mridula K. Ray; George W. Bell; Arie P. Otte; Miguel Vidal; David K. Gifford; Richard A. Young; Rudolf Jaenisch

2006-01-01

359

Bc3h1 myogenic cells produce an infectious ecotropic murine leukemia virus.  

PubMed

cDNAs representing an endogenous C-type ecotropic murine leukemia virus were isolated from a cDNA library constructed to represent mRNAs present in BC3H1 myogenic cells but not in C2C12 myogenic cells. RNA blot hybridization analysis using the cDNA inserts as probes revealed that BC3H1 cells produce MuLV-related transcirpts of at least three different size classes. A polymerase chain reaction enhanced assay for reverse transcriptase activity revealed the presence of reverse transcriptase in a viral pellet from medium conditioned by BC3H1 cells. A fungizone enhanced assay for syncitium formation provided further evidence of ecotropic retroviral particle production. Exposure of 3T3 cells to medium conditioned by BC3H1 cells, using conditions that facilitate infection, resulted in infection of the 3T3 cells, as confirmed by the syncitium formation assay. We conclude that BC3H1 cells produce an infectious ecotropic murine leukemia virus. Whether or not this feature of BC3H1 cells contributes to their inability to express some muscle-specific genes or to carry out myotube formation is unknown. Investigators will want to take into account that BC3H1 cells are virus producers when planning experiments that involve coculture of BC3H1 with other cell types, BC3H1 conditioned medium, retrovirally mediated transfection into BC3H1 cells, or study of the mCAT-1 amino acid transporter (the viral receptor) in BC3H1 cells. BC3H1 cells and the virus they produce may be of interest to those studying retroviral genomes and products and their effects. PMID:12534337

Sharp, Sandra B; Villalvazo, Maria; Espinosa, Alex; Damle, Sagar; Padilla, Xiomara; Hartono, John; Gonzalez, Rodolfo; Vu, Son

2002-01-01

360

Neurogenic potential of dental pulp stem cells isolated from murine incisors  

PubMed Central

Introduction Interest in the use of dental pulp stem cells (DPSC) to enhance neurological recovery following stroke and traumatic injury is increasing following successful pre-clinical studies. A murine model of autologous neural stem cell transplantation would be useful for further pre-clinical investigation of the underlying mechanisms. However, while human-derived DPSC have been well characterised, the neurogenic potential of murine DPSC (mDPSC) has been largely neglected. In this study we demonstrate neuronal differentiation of DPSC from murine incisors in vitro. Methods mDPSC were cultured under neuroinductive conditions and assessed for neuronal and glial markers and electrophysiological functional maturation. Results mDPSC developed a neuronal morphology and high expression of neural markers nestin, ßIII-tubulin and GFAP. Neurofilament M and S100 were found in lower abundance. Differentiated cells also expressed protein markers for cholinergic, GABAergic and glutaminergic neurons, indicating a mixture of central and peripheral nervous system cell types. Intracellular electrophysiological analysis revealed the presence of voltage-gated L-type Ca2+ channels in a majority of cells with neuronal morphology. No voltage-gated Na+ or K+ currents were found and the cultures did not support spontaneous action potentials. Neuronal-like networks expressed the gap junction protein, connexin 43 but this was not associated with dye coupling between adjacent cells after injection of the low-molecular weight tracers Lucifer yellow or Neurobiotin. This indicated that the connexin proteins were not forming traditional gap junction channels. Conclusions The data presented support the differentiation of mDPSC into immature neuronal-like networks.

2014-01-01

361

Antigen processing and presentation by a murine myoblast cell line.  

PubMed Central

The ability of non-professional antigen-presenting cells (APC) to process and present antigen to the immune system has been the subject of debate in autoimmunity and tumour immunology. The role of muscle cells in the processing and presentation of antigen to T cells via class I and class II MHC pathways is of increasing interest. Muscle cells are the targets of autoimmune attack in the inflammatory muscle diseases, and direct intramuscular injection of antigen-expressing DNA constructs is under scrutiny as a means of vaccination. Furthermore, the immunological properties of muscle cells are of relevance in attempts to transfer myoblasts as replacement cells in dystrophic diseases or as depot cells for the secretion of certain molecules in deficiency states. Using class I and class II MHC transfectant clones of the C2C12 myoblast cell line, myoblasts have been shown to be capable of presenting antigen to, and stimulating secretion of IL-2 by, T cell hybridomas via both of these pathways. The epitopes which are dominantly presented by professional APC after processing of native antigens we