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Georges Pointis - One of the best experts on this subject based on the ideXlab platform.
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Contraceptive steroids from pharmaceutical waste perturbate junctional communication in Sertoli Cells.
Biochimie, 2009Co-Authors: Mathieu Tramoni, Georges Pointis, Dominique Segretain, Jerome Gilleron, Diane Carette, Khadija Tahiri, Marie-therese Corvol, Jean-francois SavouretAbstract:The potential health impact of pharmaceutical waste is now a growing concern. Contraceptive steroids are prominent environmental contaminants and thus may act as endocrine disruptors. Numerous xenobiotics hamper Sertoli Cells junctional communication which is known to participate in spermatogenesis control. This has been associated with male subfertility and testicular cancer. We investigated three contraceptive molecules found in the environment for their potential impact on Sertoli Cells gap junction functionality: 17a-ethynylestradiol, medroxyprogesterone acetate and levonorgestrel. Four other non-steroid drugs also found in the environment were included in the study. Communication disruption was analyzed in vitro in murine seminiferous tubules and the 42GPA9 Sertoli Cell Line. Steroids modulated connexin43 trafficking and impaired junctional communication through rapid effects apparently acting on the Cell membrane but not on Cx43 expression. The 4 non-steroid compounds showed no effect. Longer exposure to steroids increased gap junction impairment, which was associated in part with Na/K ATPase internalization. Estrogen receptors (ER) did not appear to be involved in gap junction disruption: Sertoli Cells are devoid of ERalpha and only express the cytoplasmic beta isoform. ERbeta localization was not modified by either steroid. The threshold level was surprisingly low, around 10(-16) M. We conclude that steroidal pollutants disrupt Sertoli Cells junctional communication in vitro at concentrations that can be found in the environment.
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Three-dimensional analysis of connexin43 gap junction in the ex vivo rat seminiferous tubules: Short-term effects of hormonal effectors
Microscopy research and technique, 2009Co-Authors: Jerome Gilleron, Dominique Segretain, Diane Carette, Francoise Carpentier, Georges PointisAbstract:Cx43 gap junctions are essential for proliferation, differentiation, and apoptosis of germ Cells during spermatogenesis. However, only few and indirect observations have been reported on the distribution of Cx43, the predominant Cx within the seminiferous tubules. In thepresent study, we developed an innovative method that allows visualization of the three- dimensional localization of Cx43 associated with gap junctions and their functionality in isolated spermatogenic stage-specific seminiferous tubules. Cx43 gap junctions were present between myoid Cells, between Sertoli Cells, and between Sertoli and germ Cells. Cx43 levels and coupling were stage-dependent with higher values at stages VI–VIII of spermatogenesis and markedly reduced at stages IX–X. Short-term exposure of seminiferous tubule fragments at stages VI–VIII and of the 42GPA9 Sertoli Cell Line transfected with a Cx43-GFP vector, to FSH, cAMP, DHT, and 17β-E2 significantly altered Cx43 distribution as well as gap junction coupling. These observations highlight a nongenomic effect of these testicular effectors on Cx43 gap junction. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc.
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Accelerated internalization of junctional membrane proteins (connexin 43, N-cadherin and ZO-1) within endocytic vacuoles: An early event of DDT carcinogenicity
Biochimica et biophysica acta, 2007Co-Authors: Céline Fiorini, Stephan Chevalier, Dominique Segretain, Jerome Gilleron, Diane Carette, Astrid Valette, Anne Tilloy, Georges PointisAbstract:Abstract Stability of Cell-to-Cell interactions and integrity of junctional membrane proteins are essential for biological processes including cancer prevention. The present study shows that DDT, a non-genomic carcinogen used at a non-cytotoxic dose (1 μM), rapidly disrupted the Cell–Cell contacts and concomitantly induced the formation of cytoplasmic vacuoles close to the plasma membrane in the SerW3 Sertoli Cell Line. High-resolution deconvolution microscopy reveals that this vacuolization process was clathrin-dependent since a hyperosmotic media (0.2 M sucrose) blocked rhodamine–dextran endocytosis. In response to DDT, junctional proteins such as Cx43, N-Cadherin and ZO-1 were internalized and present in vacuoles. In Cx43-GFP transfected Cells, time lapse videomicroscopy demonstrates that DDT rapidly enhanced fragmentation of the gap junction plaques and abolished the gap junction coupling without major modification of Cx43 phosphorylation status. Repeated exposure to DDT resulted in chronic gap junction coupling injury. The present results demonstrate that one of the early effect of DDT is to interfere with the plasma membrane and to perturb its function, specifically its ability to establish Cell–Cell junctions that are essential for tissue homeostasis and control of Cell proliferation and differentiation. Such an alteration may play a specific role during carcinogenesis.
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Sertoli Cell junctional proteins as early targets for different classes of reproductive toxicants
Reproductive toxicology (Elmsford N.Y.), 2004Co-Authors: Céline Fiorini, Anne Tilloy-ellul, Stephan Chevalier, Claude Charuel, Georges PointisAbstract:In the testis, Sertoli Cells establish interCellular junctions that are essential for spermatogenesis. The SerW3 Sertoli Cell Line displays some features of native Sertoli Cells. Western blot and immunofluorescence analyses showed that SerW3 Sertoli Cells expressed typical components of tight (occludin and zonula occludens-1), anchoring (N-cadherin) and gap (connexin 43) junctions. Testicular toxicants (DDT, pentachlorophenol, dieldrin, dinitrobenzene, cadmium chloride, cisplatin, gossypol, bisphenol A and tert-octylphenol) affected interCellular junctions by either reducing the amount or inducing aberrant intraCellular localization of these membranous proteins. Phosphodiesterase inhibitors (isobutyl methylxantine, rolipram, zaprinast, zardaverine) did not alter junctional-complex component levels but caused a rapid and reversible redistribution of these proteins to the cytoplasmic compartment. The present study showed that occludin, ZO-1, N-cadherin and specifically Cx43 could be early targets for testicular toxicants. The SerW3 Cell Line therefore appears as a useful in vitro model to evaluate molecules with potential anti-reproductive effects.
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aberrant connexin 43 endocytosis by the carcinogen lindane involves activation of the erk mitogen activated protein kinase pathway
Carcinogenesis, 2003Co-Authors: Baharia Mograbi, Norah Defamie, Dominique Segretain, P Fenichel, Elisabeth Corcelle, Michel Samson, Marielle Nebout, Georges PointisAbstract:: Although worldwide concerns have emerged about environmental factors that display carcinogenic and reprotoxic effects, little is known about the mechanism(s) by which these chemicals alter testicular function. Using the 42GPA9 Sertoli Cell Line, we recently reported that one widely used lipid-soluble pesticide, Lindane impairs gap junctional interCellular communication by promoting the intraCellular localization of Connexin 43 (Cx43), a tumor suppressor. We showed here that this chemical triggered the accumulation of Cx43 within Rab5 positive endosomes. Interestingly, evidence is provided that Lindane-induced Cx43 endocytosis did not stem on alteration of Cx43 partition in lipid rafts. Lindane induced concomitantly Cx43 phosphorylation and activation of extraCellular signal-regulated kinases (ERK) but not of JNK and p38 mitogen- activated protein kinases. Inhibition of ERK pathway by PD98059, a MEK1-specific inhibitor, prevented Lindane-induced Cx43 phosphorylation, restored Cx43 membranous localization and gap junction coupling. Altogether, these findings provide the first evidence that Lindane-altered Cx43 endocytosis requires ERK activation. Such inappropriate activation of the mitogenic MAPK pathway and inactivation of the tumor suppressor Cx43 by Lindane may participate in the promotion of neoplastic Cell growth.
Masuo Obinata - One of the best experts on this subject based on the ideXlab platform.
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pspc1 nono and sfpq are expressed in mouse Sertoli Cells and may function as coregulators of androgen receptor mediated transcription
Biology of Reproduction, 2006Co-Authors: Sho Kuwahara, Yoshiaki Tabuchi, Masuo Obinata, Asako Ikei, Yusuke Taguchi, Nariaki Fujimoto, Seiichi Uesugi, Yasuyuki KuriharaAbstract:In Sertoli Cells of testis, androgen receptor-regulated gene transcription plays an indispensable role in maintaining spermatogenesis. Androgen receptor activity is modulated by a number of coregulators which are associated with the androgen receptor. Non-POU-domain-containing, octamer binding protein (NONO), a member of the DBHS-containing proteins, complexes with androgen receptor and functions as a coactivator for the receptor. Paraspeckle protein 1 alpha isoform (PSPC1, previously known as PSP1) and Splicing factor, proLine- and glutamine-rich (SFPQ, previously known as PSF), other members of the DBHS-containing proteins, are also found in androgen receptor complexes, suggesting that these DBHS-containing proteins may cooperatively regulate androgen receptor-mediated gene transcription. We demonstrated that PSPC1, NONO, and SFPQ are coexpressed in Sertoli Cell Line TTE3 and interact reciprocally. The effect of the DBHS-containing proteins on the transcriptional activity was assessed using the construct containing androgen-responsive elements followed by a luciferase gene. The results showed that all the DBHS-containing proteins activate androgen receptor-mediated transcription, and PSPC1 is the most effective coactivator among them. Furthermore, we confirmed the presence of PSPC1, NONO, and SFPQ proteins in Sertoli Cells of adult mouse testis sections. These observations suggest that PSPC1, NONO, and SFPQ form complexes with each other in Sertoli Cells and may regulate androgen receptor-mediated transcriptional activity.
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Development of a conditionally immortalized testicular Sertoli Cell Line RTS3-3 from adult transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene.
Cell structure and function, 2003Co-Authors: Yoshiaki Tabuchi, Ri-ichi Takahashi, Masatsugu Ueda, Masuo ObinataAbstract:Transgenic mice and rats harboring temperature-sensitive simian virus 40 (tsSV40) large T-antigen gene are useful for establishing Cell Lines from tissues. We succeeded in establishing a conditionally immortalized testicular Sertoli Cell Line, RT3-3, from adult transgenic rats harboring the oncogene. The Cells grew at permissive (33°C) and intermediate (37°C) temperatures but not at nonpermissive temperature (39°C). Large T-antigen was expressed at 33 and 37°C, whereas the expression level was gradually decreased at 39°C, suggesting that the temperature-sensitive growth characteristics arise as a result of the function of tsSV40 large T-antigen. The Cells showed biochemical features associate with normal Sertoli Cells including expressions of mRNAs of sulfated glycoprotein-2 (SGP-2), transferrin (TF) and steel factor. Quantitative polymerase chain reaction revealed that nonpermissive temperature induced increase in the level of SGP-2. Moreover, levels of SGP-2 and/or TF were significantly elevated in the Cells treatment with sodium butyrate and retinoic acid, inducers of Cellular differentiation. To our knowledge, this is the first report of the establishment of a testicular Sertoli Cell Line from the transgenic rats. Thus, the conditionally immortalized Cell Line RTS3-3 with unique characteristics may serve as good experimental in vitro models for basic and applied biology of testicular Sertoli Cells.
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DEVELOPMENT OF THE CONDITIONALLY IMMORTALIZED TESTICULAR Sertoli Cell Line TTE3 EXPRESSING Sertoli Cell SPECIFIC GENES FROM MICE TRANSGENIC FOR TEMPERATURE SENSITIVE SIMIAN VIRUS 40 LARGE T ANTIGEN GENE
The Journal of urology, 2002Co-Authors: Yoshiaki Tabuchi, Shoichiro Ohta, Nobuaki Yanai, Masuo Obinata, Takashi Kondo, Hideki Fuse, Shinji AsanoAbstract:Purpose: We developed and characterized a conditionally immortalized testicular Sertoli Cell Line from transgenic mice bearing the temperature sensitive simian virus 40 large T antigen gene pSVtsA58.Materials and Methods: Established Cells from 8-week-old male transgenic mice were cultured at a permissive (33C) or nonpermissive (39C) temperature on a collagen type I pre-coated culture vessel. The expression of Sertoli Cell specific proteins was analyzed by reverse transcriptase-polymerase chain reaction, immunocytochemical testing and Western blot analysis.Results: The Sertoli Cell Line TTE3 grew at 33C but not at 39C. Large T antigen was expressed only in the nuclei at 33C, indicating that the temperature sensitive growth phenotype of the Cells arose as a result of the function of temperature sensitive simian virus 40 large T antigen. The Cells did not show any colony forming activity in soft agar or form tumors in subcutaneous tissue in nude mice, showing that TTE3 Cells were not transformed. The Cells ...
Yoshiaki Tabuchi - One of the best experts on this subject based on the ideXlab platform.
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pspc1 nono and sfpq are expressed in mouse Sertoli Cells and may function as coregulators of androgen receptor mediated transcription
Biology of Reproduction, 2006Co-Authors: Sho Kuwahara, Yoshiaki Tabuchi, Masuo Obinata, Asako Ikei, Yusuke Taguchi, Nariaki Fujimoto, Seiichi Uesugi, Yasuyuki KuriharaAbstract:In Sertoli Cells of testis, androgen receptor-regulated gene transcription plays an indispensable role in maintaining spermatogenesis. Androgen receptor activity is modulated by a number of coregulators which are associated with the androgen receptor. Non-POU-domain-containing, octamer binding protein (NONO), a member of the DBHS-containing proteins, complexes with androgen receptor and functions as a coactivator for the receptor. Paraspeckle protein 1 alpha isoform (PSPC1, previously known as PSP1) and Splicing factor, proLine- and glutamine-rich (SFPQ, previously known as PSF), other members of the DBHS-containing proteins, are also found in androgen receptor complexes, suggesting that these DBHS-containing proteins may cooperatively regulate androgen receptor-mediated gene transcription. We demonstrated that PSPC1, NONO, and SFPQ are coexpressed in Sertoli Cell Line TTE3 and interact reciprocally. The effect of the DBHS-containing proteins on the transcriptional activity was assessed using the construct containing androgen-responsive elements followed by a luciferase gene. The results showed that all the DBHS-containing proteins activate androgen receptor-mediated transcription, and PSPC1 is the most effective coactivator among them. Furthermore, we confirmed the presence of PSPC1, NONO, and SFPQ proteins in Sertoli Cells of adult mouse testis sections. These observations suggest that PSPC1, NONO, and SFPQ form complexes with each other in Sertoli Cells and may regulate androgen receptor-mediated transcriptional activity.
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Development of a conditionally immortalized testicular Sertoli Cell Line RTS3-3 from adult transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene.
Cell structure and function, 2003Co-Authors: Yoshiaki Tabuchi, Ri-ichi Takahashi, Masatsugu Ueda, Masuo ObinataAbstract:Transgenic mice and rats harboring temperature-sensitive simian virus 40 (tsSV40) large T-antigen gene are useful for establishing Cell Lines from tissues. We succeeded in establishing a conditionally immortalized testicular Sertoli Cell Line, RT3-3, from adult transgenic rats harboring the oncogene. The Cells grew at permissive (33°C) and intermediate (37°C) temperatures but not at nonpermissive temperature (39°C). Large T-antigen was expressed at 33 and 37°C, whereas the expression level was gradually decreased at 39°C, suggesting that the temperature-sensitive growth characteristics arise as a result of the function of tsSV40 large T-antigen. The Cells showed biochemical features associate with normal Sertoli Cells including expressions of mRNAs of sulfated glycoprotein-2 (SGP-2), transferrin (TF) and steel factor. Quantitative polymerase chain reaction revealed that nonpermissive temperature induced increase in the level of SGP-2. Moreover, levels of SGP-2 and/or TF were significantly elevated in the Cells treatment with sodium butyrate and retinoic acid, inducers of Cellular differentiation. To our knowledge, this is the first report of the establishment of a testicular Sertoli Cell Line from the transgenic rats. Thus, the conditionally immortalized Cell Line RTS3-3 with unique characteristics may serve as good experimental in vitro models for basic and applied biology of testicular Sertoli Cells.
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DEVELOPMENT OF THE CONDITIONALLY IMMORTALIZED TESTICULAR Sertoli Cell Line TTE3 EXPRESSING Sertoli Cell SPECIFIC GENES FROM MICE TRANSGENIC FOR TEMPERATURE SENSITIVE SIMIAN VIRUS 40 LARGE T ANTIGEN GENE
The Journal of urology, 2002Co-Authors: Yoshiaki Tabuchi, Shoichiro Ohta, Nobuaki Yanai, Masuo Obinata, Takashi Kondo, Hideki Fuse, Shinji AsanoAbstract:Purpose: We developed and characterized a conditionally immortalized testicular Sertoli Cell Line from transgenic mice bearing the temperature sensitive simian virus 40 large T antigen gene pSVtsA58.Materials and Methods: Established Cells from 8-week-old male transgenic mice were cultured at a permissive (33C) or nonpermissive (39C) temperature on a collagen type I pre-coated culture vessel. The expression of Sertoli Cell specific proteins was analyzed by reverse transcriptase-polymerase chain reaction, immunocytochemical testing and Western blot analysis.Results: The Sertoli Cell Line TTE3 grew at 33C but not at 39C. Large T antigen was expressed only in the nuclei at 33C, indicating that the temperature sensitive growth phenotype of the Cells arose as a result of the function of temperature sensitive simian virus 40 large T antigen. The Cells did not show any colony forming activity in soft agar or form tumors in subcutaneous tissue in nude mice, showing that TTE3 Cells were not transformed. The Cells ...
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DNA microarray analysis of differentially expressed genes responsive to bisphenol A, an alkylphenol derivative, in an in vitro mouse Sertoli Cell model.
Japanese journal of pharmacology, 2002Co-Authors: Yoshiaki Tabuchi, Qing-li Zhao, Takashi KondoAbstract:To identify genes elicited by bisphenol A (BPA) in Sertoli Cells, we carried out a microarray analysis of TTE3 Cells (a mouse Sertoli Cell Line) treated with BPA. BPA (100, 200 and 400 μM) induced Cell death concentration-dependently, with levels being 25%, 33% and 96%, respectively. Of the 1,081 genes analyzed, 3 genes showed decreased levels of expression while the remaining 10 genes showed increased levels in the Cells treated with a subtoxic dose of BPA (200 μM). The expressions of six genes were confirmed by the TaqMan assay. These findings suggest that DNA microarray analysis is a useful tool for investigating the molecular mechanisms of the toxic effects of BPA in testicular Cells.
Michael D. Griswold - One of the best experts on this subject based on the ideXlab platform.
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expression of clusterin sulfated glycoprotein 2 under conditions of heat stress in rat Sertoli Cells and a mouse Sertoli Cell Line
Journal of Andrology, 1997Co-Authors: Ann M. Clark, Michael D. GriswoldAbstract:Clusterin is the major protein produced by rat Sertoli Cells and is deposited onto sperm membranes; however, its function is unknown. In order to gain insight into the regulation of clusterin in Sertoli Cells, the objective of the present study was to develop a model where the expression of clusterin could be affected in Sertoli Cells in vitro. Rat Sertoli Cells and mouse Sertoli Cells (MSC1) were cultured under heat stress conditions (41 degrees C) for up to 48 hours. The mRNA for clusterin in Sertoli Cells was compared to that in human epitheliod cancer Cells (A431) to determine if clusterin expression was regulated in a lestis-specific manner. The mRNA for heat shock protein 70 (HSP70) was also examined as it is a known stress-regulated gene. Expression of HSP70 mRNA was increased in all three Cell types by 4 hours after the start of heat stress. Clusterin mRNA was increased over that of controls by 4 hours in heat-stressed A431 Cells but did not significantly increase in MSC1 or Sertoli Cells until 12 hours (P < 0.05). The induction of clusterin mRNA in MSC1 Cells continued for at least 48 hours and required the sustained exposure of Cells to the 41 degrees C temperature. The increase in the amount of clusterin mRNA was not due to an increase in transcript half-life, as determined by the addition of actinomycin D to the media of control vs. heat-stressed MSC1 Cells. From the development of this in vitro model, we have seen that the timing of induction of clusterin by heat stress is Sertoli Cell specific and is different than that of HSP70. This response in surviving Cells during heat stress may be protective in that clusterin would bind to toxic compounds or solubilize Cellular debris released by degenerating Cells.
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Expression of Clusterin/Sulfated Glycoprotein-2 Under Conditions of Heat Stress in Rat Sertoli Cells and a Mouse Sertoli Cell Line
Journal of andrology, 1997Co-Authors: Ann M. Clark, Michael D. GriswoldAbstract:Clusterin is the major protein produced by rat Sertoli Cells and is deposited onto sperm membranes; however, its function is unknown. In order to gain insight into the regulation of clusterin in Sertoli Cells, the objective of the present study was to develop a model where the expression of clusterin could be affected in Sertoli Cells in vitro. Rat Sertoli Cells and mouse Sertoli Cells (MSC1) were cultured under heat stress conditions (41 degrees C) for up to 48 hours. The mRNA for clusterin in Sertoli Cells was compared to that in human epitheliod cancer Cells (A431) to determine if clusterin expression was regulated in a lestis-specific manner. The mRNA for heat shock protein 70 (HSP70) was also examined as it is a known stress-regulated gene. Expression of HSP70 mRNA was increased in all three Cell types by 4 hours after the start of heat stress. Clusterin mRNA was increased over that of controls by 4 hours in heat-stressed A431 Cells but did not significantly increase in MSC1 or Sertoli Cells until 12 hours (P < 0.05). The induction of clusterin mRNA in MSC1 Cells continued for at least 48 hours and required the sustained exposure of Cells to the 41 degrees C temperature. The increase in the amount of clusterin mRNA was not due to an increase in transcript half-life, as determined by the addition of actinomycin D to the media of control vs. heat-stressed MSC1 Cells. From the development of this in vitro model, we have seen that the timing of induction of clusterin by heat stress is Sertoli Cell specific and is different than that of HSP70. This response in surviving Cells during heat stress may be protective in that clusterin would bind to toxic compounds or solubilize Cellular debris released by degenerating Cells.
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Relationship of a mouse Sertoli Cell Line (MSC-1) to normal Sertoli Cells.
Biology of reproduction, 1994Co-Authors: Michael Mcguinness, Carol C. Linder, Carlos R. Morales, Leslie L. Heckert, Jeremie Pikus, Michael D. GriswoldAbstract:Recently, a mouse Sertoli Cell Line (MSC- I) was established from transgenic mice carrying a fusion gene composed of human Muillerian inhibitory substance transcriptional regulatory sequences linked to the SV40 T-antigen gene. This Cell Line contained a morphologically heterogeneous population of Cells that expressed characteristic Sertoli Cell mRNAs such as transferrin, the Psubunit for inhibin, and sulfated glycoprotein-2 (SGP-2). In this study, we compared various characteristics of MSC-1 Cells to primary cultures of Sertoli Cells from immature rats or adult mice. Our observations indicated that: 1) These Cells were ultrastructurally similar to mouse Sertoli Cells in culture; 2) MSC-1 Cells expressed mRNA for androgen-binding protein (ABP) and SGP-1, but not the receptor for FSH; 3) The expression of SGP-2 mRNA and secretion of SGP-2 increased approximately 2-fold when Cells were cultured at 41 0 C, the nonpermissive temperature for the SV40 virus, while SGP-1 and transferrin mRNA levels and secretion were unaffected; 4) Proliferation of these Cells was serum-dose dependent and temperature dependent and could be inhibited by incubating MSC-1 Cells at 41 0 C; 5) Proliferation was also significantly reduced after Cells were incubated in the presence of dibutyryl cAMP (dbcAMP) for 6 days; 6) Fifteen subcultures produced from single MSC-1 Cells displayed similar levels of mRNA expression for transferrin or SGP-1. Together these data indicate that the MSC-1 Cell Line is composed of a single Cell type displaying numerous characteristics of Sertoli Cells. This Cell Line may provide a unique source of tissue for studying various aspects of Sertoli Cell behavior.
Shinji Asano - One of the best experts on this subject based on the ideXlab platform.
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DEVELOPMENT OF THE CONDITIONALLY IMMORTALIZED TESTICULAR Sertoli Cell Line TTE3 EXPRESSING Sertoli Cell SPECIFIC GENES FROM MICE TRANSGENIC FOR TEMPERATURE SENSITIVE SIMIAN VIRUS 40 LARGE T ANTIGEN GENE
The Journal of urology, 2002Co-Authors: Yoshiaki Tabuchi, Shoichiro Ohta, Nobuaki Yanai, Masuo Obinata, Takashi Kondo, Hideki Fuse, Shinji AsanoAbstract:Purpose: We developed and characterized a conditionally immortalized testicular Sertoli Cell Line from transgenic mice bearing the temperature sensitive simian virus 40 large T antigen gene pSVtsA58.Materials and Methods: Established Cells from 8-week-old male transgenic mice were cultured at a permissive (33C) or nonpermissive (39C) temperature on a collagen type I pre-coated culture vessel. The expression of Sertoli Cell specific proteins was analyzed by reverse transcriptase-polymerase chain reaction, immunocytochemical testing and Western blot analysis.Results: The Sertoli Cell Line TTE3 grew at 33C but not at 39C. Large T antigen was expressed only in the nuclei at 33C, indicating that the temperature sensitive growth phenotype of the Cells arose as a result of the function of temperature sensitive simian virus 40 large T antigen. The Cells did not show any colony forming activity in soft agar or form tumors in subcutaneous tissue in nude mice, showing that TTE3 Cells were not transformed. The Cells ...
