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Published in: Reproductive Biology and Endocrinology 1/2003

Open Access 01-12-2003 | Research

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

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

Published in: Reproductive Biology and Endocrinology | Issue 1/2003

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Abstract

Several in vivo studies have reported the presence of immunoreactive transforming growth factor-β's (TGF-β's) in testicular cells at defined stages of their differentiation. The most pronounced changes in TGF-β1 and TGF-β2 immunoreactivity occurred during spermatogenesis. In the present study we have investigated whether germ cells and Sertoli cells are able to secrete bioactive TGF-β's in vitro, using the CCl64 mink lung epithelial cell line as bioassay for the measurement of TGF-β. In cellular lysates, TGF-β bioactivity was only observed following heat-treatment, indicating that within these cells TGF-β is present in a latent form. To our surprise, active TGF-β 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-β in a latent form, but that they also release a factor which can convert latent TGF-β into its active form. Following heat-activation of these culture supernatant's, total TGF-β bioactivity increased 6- to 9-fold. Spermatocytes are the cell type that releases most bioactive TGF-β during a 24 h culture period, although round and elongated spermatids and Sertoli cells also secrete significant amounts of TGF-β. The biological activity of TGF-β could be inhibited by neutralizing antibodies against TGF-β1 (spermatocytes and round spermatids) and TGF-β2 (round and elongating spermatids). TGF-β activity in the Sertoli cell culture supernatant was inhibited slightly by either the TGF-β1 and TGF-β2 neutralizing antibody.
These in vitro data suggest that germ cells and Sertoli cells release latent TGF-β's. Following secretion, the TGF-β's are converted to a biological active form that can interact with specific TGF-β receptors. These results strengthen the hypothesis that TGF-β's may play a physiological role in germ cell proliferation/differentiation and Sertoli cell function.
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Literature
1.
go back to reference Bellvé AR, Zheng W: Growth factors as autocrine and paracrine modulators of male gonadale functions. J Reprod Fertil. 1989, 85: 771-793.CrossRefPubMed Bellvé AR, Zheng W: Growth factors as autocrine and paracrine modulators of male gonadale functions. J Reprod Fertil. 1989, 85: 771-793.CrossRefPubMed
2.
go back to reference Avallet O, Vigier M, Albaladejo V, de Cesaris P, Saez JM: Transforming growth factor β gene expression in cultured porcine Sertoli and Leydig cells: Effects of hormine and growth factors. In: Proceedings of the 6th European Workshop on Molecular and Cellular Endocrinology of the Testis. 1990, D13: Avallet O, Vigier M, Albaladejo V, de Cesaris P, Saez JM: Transforming growth factor β gene expression in cultured porcine Sertoli and Leydig cells: Effects of hormine and growth factors. In: Proceedings of the 6th European Workshop on Molecular and Cellular Endocrinology of the Testis. 1990, D13:
3.
go back to reference Caussanel S, Tabone E, Hendrick JC, Dacheux F, Benahmed M: Cellular distribution of transforming growth factor betas 1, 2, and 3 and their types I and II receptors during postnatal development and spermatogenesis in the boar testis. Biol Reprod. 1997, 56: 357-367.CrossRefPubMed Caussanel S, Tabone E, Hendrick JC, Dacheux F, Benahmed M: Cellular distribution of transforming growth factor betas 1, 2, and 3 and their types I and II receptors during postnatal development and spermatogenesis in the boar testis. Biol Reprod. 1997, 56: 357-367.CrossRefPubMed
4.
go back to reference Watrin F, Scotto L, Associan RK, Wolgemuth DJ: Cell lineage specificity of expression of the murine transforming growth factor-β3 and transforming growth factor-β1 genes. Cell Growth & Differ. 1991, 2: 77-83. Watrin F, Scotto L, Associan RK, Wolgemuth DJ: Cell lineage specificity of expression of the murine transforming growth factor-β3 and transforming growth factor-β1 genes. Cell Growth & Differ. 1991, 2: 77-83.
5.
go back to reference Skinner MK, Moses HL: Transforming growth factorβ gene expression and action in the seminiferous tubule: Peritubular cell-Sertoli cell interactions. Mol Endocrinol. 1989, 3: 625-634.CrossRefPubMed Skinner MK, Moses HL: Transforming growth factorβ gene expression and action in the seminiferous tubule: Peritubular cell-Sertoli cell interactions. Mol Endocrinol. 1989, 3: 625-634.CrossRefPubMed
6.
go back to reference Mullaney BP, Skinner MK: Transforming growth factor-β (β1, β2, and β3) gene expression and action during pubertal development of the seminiferous tubule: Potential role at the onset of spermatogenesis. Mol Endocrinol. 1993, 7: 67-76.PubMed Mullaney BP, Skinner MK: Transforming growth factor-β (β1, β2, and β3) gene expression and action during pubertal development of the seminiferous tubule: Potential role at the onset of spermatogenesis. Mol Endocrinol. 1993, 7: 67-76.PubMed
7.
go back to reference Teerds KJ, Dorrington JH: Localization of transforming growth factor β1 and β2 during testicular development in the rat. Biol Reprod. 1993, 48: 40-45.CrossRefPubMed Teerds KJ, Dorrington JH: Localization of transforming growth factor β1 and β2 during testicular development in the rat. Biol Reprod. 1993, 48: 40-45.CrossRefPubMed
8.
go back to reference Themmen APN, Blok LJ, Post M, Baarends WM, Hoogerbrugge JW, Parmentier M, Vassert G, Grootegoed JA: Follitropin receptor down-regulation involves a cAMP-dependent post-transcriptional decrease of receptor mRNA expression. Molec Cell Endocrinol. 1991, 78: R7-R13.CrossRefPubMed Themmen APN, Blok LJ, Post M, Baarends WM, Hoogerbrugge JW, Parmentier M, Vassert G, Grootegoed JA: Follitropin receptor down-regulation involves a cAMP-dependent post-transcriptional decrease of receptor mRNA expression. Molec Cell Endocrinol. 1991, 78: R7-R13.CrossRefPubMed
9.
go back to reference Grootegoed JA, Jansen R, van der Molen HJ: Effect of glucose on ATP dephosphorylation in rat spermatids. J Reprod Fertil. 1986, 77: 99-107.CrossRefPubMed Grootegoed JA, Jansen R, van der Molen HJ: Effect of glucose on ATP dephosphorylation in rat spermatids. J Reprod Fertil. 1986, 77: 99-107.CrossRefPubMed
10.
go back to reference Toebosch AMW, Brussée R, Verkerk A, Grootegoed JA: Quantitative evaluation of the maintenance and development of spermatocytes and round spermatids in cultured tubule fragments from immature rat testis. Int J Androl. 1989, 12: 360-374.CrossRefPubMed Toebosch AMW, Brussée R, Verkerk A, Grootegoed JA: Quantitative evaluation of the maintenance and development of spermatocytes and round spermatids in cultured tubule fragments from immature rat testis. Int J Androl. 1989, 12: 360-374.CrossRefPubMed
11.
go back to reference Jutte NHPM, Jansen R, Grootegoed JA, Rommerts FFG, van der Molen HJ: Protein synthesis by isolated pachytene spermatocytes in the absence of Sertoli cells. J Exp Zool. 1985, 233: 285-290.CrossRefPubMed Jutte NHPM, Jansen R, Grootegoed JA, Rommerts FFG, van der Molen HJ: Protein synthesis by isolated pachytene spermatocytes in the absence of Sertoli cells. J Exp Zool. 1985, 233: 285-290.CrossRefPubMed
12.
go back to reference Grootegoed JA, Grollé-Hey AH, Rommerts FFG, van der Molen HJ: Ribonucleic acid synthesis in vitro in primary spermatocytes isolated from rat testis. Biochem J. 1977, 168: 23-31.PubMedCentralCrossRefPubMed Grootegoed JA, Grollé-Hey AH, Rommerts FFG, van der Molen HJ: Ribonucleic acid synthesis in vitro in primary spermatocytes isolated from rat testis. Biochem J. 1977, 168: 23-31.PubMedCentralCrossRefPubMed
13.
go back to reference Grootegoed JA, Krüger-Sewnarain RC, Jutte NHPM, Rommerts FFG, van der Molen HJ: Fucosylation of glycoproteins in rat spermatocytes and spermatids. Gam Res. 1982, 5: 303-315.CrossRef Grootegoed JA, Krüger-Sewnarain RC, Jutte NHPM, Rommerts FFG, van der Molen HJ: Fucosylation of glycoproteins in rat spermatocytes and spermatids. Gam Res. 1982, 5: 303-315.CrossRef
14.
go back to reference Danielpour D, Dart LL, Flanders KC, Roberts AB, Sporn MB: Immunodetection and quantitation of the two forms of transforming growth factor-β (TGF-β1 and TGF-β2) secreted by cells in culture. J Cell Physiol. 1989, 138: 79-86.CrossRefPubMed Danielpour D, Dart LL, Flanders KC, Roberts AB, Sporn MB: Immunodetection and quantitation of the two forms of transforming growth factor-β (TGF-β1 and TGF-β2) secreted by cells in culture. J Cell Physiol. 1989, 138: 79-86.CrossRefPubMed
15.
go back to reference Kalkhoven E, Kwakkenbos-Isbrucker L, Mummery CL, de Laat SW, van den Eijnden-van Raaij AJ, van der Saag PT, van den Burg B: The role of TGF-beta production in growth inhibition of breast-tumor cells by progestins. Int J Cancer. 1995, 61: 80-86.CrossRefPubMed Kalkhoven E, Kwakkenbos-Isbrucker L, Mummery CL, de Laat SW, van den Eijnden-van Raaij AJ, van der Saag PT, van den Burg B: The role of TGF-beta production in growth inhibition of breast-tumor cells by progestins. Int J Cancer. 1995, 61: 80-86.CrossRefPubMed
16.
go back to reference Sporn MB, Roberts AB, Wakefield LM, De Crombrugge B: Some recent advances in the chemistry and biology of transforming growth factor-β. J Cell Biol. 1987, 105: 1039-1045.CrossRefPubMed Sporn MB, Roberts AB, Wakefield LM, De Crombrugge B: Some recent advances in the chemistry and biology of transforming growth factor-β. J Cell Biol. 1987, 105: 1039-1045.CrossRefPubMed
17.
go back to reference Avallet O, Gomez E, Vigier M, Jégou B, Saez JM: Sertoli cell-germ cell interactions and TGFβ1 expression and secretion in vitro. Biochem Biophys Res Comm. 1997, 238: 905-909.CrossRefPubMed Avallet O, Gomez E, Vigier M, Jégou B, Saez JM: Sertoli cell-germ cell interactions and TGFβ1 expression and secretion in vitro. Biochem Biophys Res Comm. 1997, 238: 905-909.CrossRefPubMed
18.
go back to reference Takiuchi H, Tada T, Li XF, Ogata M, Ikeda T, Fujimoto S, Fujiwara H, Hamaoka T: Particular types of tumor cells have the capacity to convert transforming growth factor β from a latent to an active form. Cancer Res. 1992, 52: 5641-5646.PubMed Takiuchi H, Tada T, Li XF, Ogata M, Ikeda T, Fujimoto S, Fujiwara H, Hamaoka T: Particular types of tumor cells have the capacity to convert transforming growth factor β from a latent to an active form. Cancer Res. 1992, 52: 5641-5646.PubMed
19.
go back to reference Sporn MB, Roberts AB, Wakefield LM, Associan RK: Transforming growth factor β: Biological function and chemical structure. Science. 1986, 233: 532-534.CrossRefPubMed Sporn MB, Roberts AB, Wakefield LM, Associan RK: Transforming growth factor β: Biological function and chemical structure. Science. 1986, 233: 532-534.CrossRefPubMed
20.
go back to reference Pircher R, Jullien P, Lawrence DA: β-Transforming growth factor is stored in human blood platelets as a latent high molecular weight complex. Biochem Biophys Res Commun. 1986, 136: 30-37.CrossRefPubMed Pircher R, Jullien P, Lawrence DA: β-Transforming growth factor is stored in human blood platelets as a latent high molecular weight complex. Biochem Biophys Res Commun. 1986, 136: 30-37.CrossRefPubMed
21.
go back to reference Knabbe C, Lippman ME, Wakefield LM, Flanders KC, Kasid A, Derynck R, Dickson RB: Evidence that transforming growth factor β is a hormonally regulated negative growth factor in human breast cancer cells. Cell. 1987, 48: 417-428.CrossRefPubMed Knabbe C, Lippman ME, Wakefield LM, Flanders KC, Kasid A, Derynck R, Dickson RB: Evidence that transforming growth factor β is a hormonally regulated negative growth factor in human breast cancer cells. Cell. 1987, 48: 417-428.CrossRefPubMed
22.
go back to reference Associan RK, Fleurdelys BE, Stevenson HC, Miller PJ, Madtes DK, Rains EW, Ross R, Sporn MB: Expression and secretion of type β transforming growth factor by activated human macrophages. Proc Natl Acad Sci USA. 1987, 84: 6020-6024.CrossRef Associan RK, Fleurdelys BE, Stevenson HC, Miller PJ, Madtes DK, Rains EW, Ross R, Sporn MB: Expression and secretion of type β transforming growth factor by activated human macrophages. Proc Natl Acad Sci USA. 1987, 84: 6020-6024.CrossRef
23.
go back to reference Bendell JJ, Dorrington J: Estradiol-17β stimulates DNA synthsis in rat granulosa cells: Action mediated by transforming growth factor-β. Endocrinology. 1991, 128: 2663-2665.CrossRefPubMed Bendell JJ, Dorrington J: Estradiol-17β stimulates DNA synthsis in rat granulosa cells: Action mediated by transforming growth factor-β. Endocrinology. 1991, 128: 2663-2665.CrossRefPubMed
24.
go back to reference Wakefield L: Growth factors: An overview. In: Hormonal Communicating Events in the Testis. Edited by: Isidori A, Fabbri A, Dufau ML. 1990, 181-190. Wakefield L: Growth factors: An overview. In: Hormonal Communicating Events in the Testis. Edited by: Isidori A, Fabbri A, Dufau ML. 1990, 181-190.
25.
go back to reference Morera AM, Esposito G, Ghiglieri C, Chauvin MA, Hartmann DJ, Benahmed M: Transforming growth factor β1 inhibits gonadotropin action in cultured porcine Sertoli cells. Endocrinology. 1992, 130: 831-836.PubMed Morera AM, Esposito G, Ghiglieri C, Chauvin MA, Hartmann DJ, Benahmed M: Transforming growth factor β1 inhibits gonadotropin action in cultured porcine Sertoli cells. Endocrinology. 1992, 130: 831-836.PubMed
26.
go back to reference Benahmed M, Cochet C, Keramides M, Chauvin MA, Morera AM: Evidence for a FSH dependent secretion of a receptor reactive transforming growth factor-β-like material by immature Sertoli cells in primary culture. Biochem Biophys Res Commun. 1988, 154: 1222-1231.CrossRefPubMed Benahmed M, Cochet C, Keramides M, Chauvin MA, Morera AM: Evidence for a FSH dependent secretion of a receptor reactive transforming growth factor-β-like material by immature Sertoli cells in primary culture. Biochem Biophys Res Commun. 1988, 154: 1222-1231.CrossRefPubMed
27.
go back to reference Le Magueresse-Battistoni B, Morera AM, Goddard I, Benahmed M: Expression of mRNAs for transforming growth factor-beta receptors in the rat testis. Endocrinology. 1995, 136: 2788-2791.PubMed Le Magueresse-Battistoni B, Morera AM, Goddard I, Benahmed M: Expression of mRNAs for transforming growth factor-beta receptors in the rat testis. Endocrinology. 1995, 136: 2788-2791.PubMed
28.
go back to reference Espositi G, Keramidas M, Mauduit C, Feige JJ, Morera AM, Benahmed M: Direct regulating effects of transforming growth factor beta 1 on lactate production in cultured porcine Sertoli cells. Endocrinology. 1991, 128: 1441-1449.CrossRef Espositi G, Keramidas M, Mauduit C, Feige JJ, Morera AM, Benahmed M: Direct regulating effects of transforming growth factor beta 1 on lactate production in cultured porcine Sertoli cells. Endocrinology. 1991, 128: 1441-1449.CrossRef
29.
go back to reference Panthou P, Barbey P, Thiebot B, Bocquet J: Effects of transforming growth factor-beta1, interleukin-1 alpha and interleukin-6 on rat Sertoli cell proteoglycan synthesis. Biochem Mol Biol Int. 1994, 34: 603-612.PubMed Panthou P, Barbey P, Thiebot B, Bocquet J: Effects of transforming growth factor-beta1, interleukin-1 alpha and interleukin-6 on rat Sertoli cell proteoglycan synthesis. Biochem Mol Biol Int. 1994, 34: 603-612.PubMed
30.
go back to reference W-Lui Y, Lee WM, Cheng CY: Transforming growth factor-β3 perturbs the inter-Sertoli cell tight junction permeability barrier in vitro possibly mediated via its effects on occludin, zonula occludens-1, and claudin-11. Endocrinology. 2001, 142: 1865-1877. W-Lui Y, Lee WM, Cheng CY: Transforming growth factor-β3 perturbs the inter-Sertoli cell tight junction permeability barrier in vitro possibly mediated via its effects on occludin, zonula occludens-1, and claudin-11. Endocrinology. 2001, 142: 1865-1877.
31.
go back to reference Harkovirta H, Kaipia A, Soder O, Parvinnen M: Effects of avidin-A, inhibin-A, and transforming growth factor α1 on stage-specific deoxyribonucleic acid synthesis during rat seminiferous epithelial cycle. Endocrinology. 1993, 133: 1664-1668. Harkovirta H, Kaipia A, Soder O, Parvinnen M: Effects of avidin-A, inhibin-A, and transforming growth factor α1 on stage-specific deoxyribonucleic acid synthesis during rat seminiferous epithelial cycle. Endocrinology. 1993, 133: 1664-1668.
32.
go back to reference Olaso R, Pairault C, Habert R: Expression of type I and II receptors for transforming growth factor β in the adult rat testis. Histochem Cell Biol. 1998, 110: 613-618.CrossRefPubMed Olaso R, Pairault C, Habert R: Expression of type I and II receptors for transforming growth factor β in the adult rat testis. Histochem Cell Biol. 1998, 110: 613-618.CrossRefPubMed
Metadata
Title
Rat testicular germ cells and sertoli cells release different types of bioactive transforming growth factor beta in vitro
Authors
Bart L Haagmans
Jos W Hoogerbrugge
Axel PN Themmen
Katja J Teerds
Publication date
01-12-2003
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2003
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/1477-7827-1-3

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