Metformin counteracts the effects of FSH on rat Sertoli cell proliferation

Metformin (MET) is one of the most widely used anti-hyperglycemic agents for treating patients with type 2 diabetes and it has started to be used in pediatric population at ages when Sertoli cells are still proliferating. It is well known that follicle-stimulating hormone (FSH) is the major Sertoli cell mitogen. The aim of the study is to investigate a possible effect of MET, which has been shown to have anti-proliferative properties, on FSH regulation of postnatal Sertoli cell proliferation and on the molecular mechanisms involved in this regulation. The present study was performed in eight-day-old rat Sertoli cell cultures. The results obtained show that MET in the presence of FSH increases phosphorylated acetyl-CoA carboxylase and decreases phosphorylated p70S6K levels. Moreover, we show that MET decreases FSH-stimulated Sertoli cell proliferation, and this decrease is accompanied by a reduction in FSH-stimulated Ccnd1 and Ccnd2 expression and an increase in cell cycle inhibitor p21Cip expression. Altogether, these results suggest that MET can, at least in part, counteract the effect of FSH on postnatal Sertoli cell proliferation.

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Signal transduction pathways in FSH regulation of rat Sertoli cell proliferation

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00477.2011 ◽

2012 ◽

Vol 302 (8) ◽

pp. E914-E923 ◽

Cited By ~ 49

Author(s):

María F. Riera ◽

Mariana Regueira ◽

María N. Galardo ◽

Eliana H. Pellizzari ◽

Silvina B. Meroni ◽

...

Keyword(s):

Signal Transduction ◽

Cell Proliferation ◽

Sertoli Cell ◽

Sertoli Cells ◽

Kinase Inhibitors ◽

Production Capacity ◽

Signal Transduction Pathways ◽

Mtorc1 Signaling ◽

Mtorc1 Pathway ◽

Cell Mitogen

The final number of Sertoli cells reached during the proliferative periods determines sperm production capacity in adulthood. It is well known that FSH is the major Sertoli cell mitogen; however, little is known about the signal transduction pathways that regulate the proliferation of Sertoli cells. The hypothesis of this investigation was that FSH regulates proliferation through a PI3K/Akt/mTORC1 pathway, and additionally, AMPK-dependent mechanisms counteract FSH proliferative effects. The present study was performed in 8-day-old rat Sertoli cell cultures. The results presented herein show that FSH, in addition to increasing p-Akt, p-mTOR, and p-p70S6K levels, increases p-PRAS40 levels, probably contributing to improving mTORC1 signaling. Furthermore, the decrease in FSH-stimulated p-Akt, p-mTOR, p-p70S6K, and p-PRAS40 levels in the presence of wortmannin emphasizes the participation of PI3K in FSH signaling. Additionally, the inhibition of FSH-stimulated Sertoli cell proliferation by the effect of wortmannin and rapamycin point to the relevance of the PI3K/Akt/mTORC1 signaling pathway in the mitotic activity of FSH. On the other hand, by activating AMPK, several interesting observations were made. Activation of AMPK produced an increase in Raptor phosphorylation, a decrease in p70S6K phosphorylation, and a decrease in FSH-stimulated Sertoli cell proliferation. The decrease in FSH-stimulated cell proliferation was accompanied by an increased expression of the cyclin-dependent kinase inhibitors (CDKIs) p19INK4d, p21Cip1, and p27Kip1. In summary, it is concluded that FSH regulates Sertoli cell proliferation with the participation of a PI3K/Akt/mTORC1 pathway and that AMPK activation may be involved in the detention of proliferation by, at least in part, a decrease in mTORC1 signaling and an increase in CDKI expression.

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149. Smad3 DOSAGE INFLUENCES TESTICULAR MATURATION

Reproduction Fertility and Development ◽

10.1071/srb09abs149 ◽

2009 ◽

Vol 21 (9) ◽

pp. 67

Author(s):

C. Itman ◽

C. Wong ◽

D. A. Jans ◽

M. Ernst ◽

K. L. Loveland

Keyword(s):

Cell Proliferation ◽

Sertoli Cell ◽

Sertoli Cells ◽

Post Partum ◽

Cell Maturation ◽

Physiological Status ◽

Testis Weight ◽

Testis Development ◽

Smad3 Expression ◽

Cell Mitogen

Activin A, a TGF-beta superfamily ligand which signals via Smad2 and Smad3, is critical for normal mouse testis development and quantitatively normal sperm production. Whereas activin enhances immature Sertoli cell proliferation (1), excessive activin production causes Sertoli cell tumours (2); this is alleviated when mice lack Smad3 (3). Sertoli cells exhibit developmentally regulated Smad utilization in activin signalling. Immature Sertoli cells signal via Smad3 while the onset of Smad2-mediated signal transduction correlates with Sertoli cell maturation (4). This change coincides with decreased testicular Smad3 production at puberty and a shift in follicle stimulating hormone (FSH)-induced Smad transcription, from Smad3 in 6 dpp (days post partum) Sertoli cells to Smad2 in 15 dpp cells. These findings suggest that Smad3 is more important for testis development than adult spermatogenesis. To test this hypothesis, we examined testis development in Smad3+/– and Smad3–/– mice. At 7 dpp, testis weight and cord diameter were reduced in Smad3–/–mice, indicating impaired Sertoli cell proliferation. Levels of FSH, a potent Sertoli cell mitogen, were unaltered. Histological analysis revealed advanced spermatogenesis in heterozygous mice, with round spermatids already present at 16 dpp. Quantitative PCR also identified advanced Sertoli and germ cell maturation in Smad3+/– mice, while Leydig cell maturation appeared unaltered. Adult Smad3+/– and Smad3–/– mice were fertile, but had smaller testes. This is the first study relating Smad3 levels to puberty onset and identifies the Smad3+/– mouse as a model of peripheral precocious puberty with otherwise normal physiological status, i.e. no gonadal tumours and normal FSH levels. These results demonstrate that FSH influences testis growth and maturation by regulating Smad3 expression and highlights the importance of testing whether environmental factors, toxicants and endocrine disruptors affect Smad3 expression, thereby leading to altered testis development.

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311. Adult Sertoli cells proliferate in response to exogenous follicle stimulating hormone in the adult photo-inhibited Djungarian hamster

Reproduction Fertility and Development ◽

10.1071/srb05abs311 ◽

2005 ◽

Vol 17 (9) ◽

pp. 133

Author(s):

G. A. Tarulli ◽

P. G. Stanton ◽

S. J. Meachem

Keyword(s):

Cell Proliferation ◽

Tight Junction ◽

Sertoli Cell ◽

Cell Population ◽

Sertoli Cells ◽

Follicle Stimulating Hormone ◽

Tight Junction Proteins ◽

Djungarian Hamster ◽

Junction Proteins ◽

Blood Testis Barrier

Sperm production relies on nutritional and structural support from Sertoli cells. Sertoli cells undergo maturational changes (e.g. cessation of proliferation and formation of the blood–testis barrier) around the onset of puberty in higher mammals1 and maturational failure has been associated with some infertility syndromes and testicular malignancies2. The Sertoli cell population is considered to be stable and unmodifiable by hormones after puberty in mammals, although recent data using the adult Djungarian hamster showed that Sertoli cell numbers decreased by 35% in the absence of serum gonadotrophins, and returned to control levels by short-term replacement of follicle stimulating hormone (FSH)3. Therefore, the aims of this study were to (i) quantify the proliferative activity of Sertoli cells in the hormonally manipulated Djungarian hamster, and (ii) examine the localisation of several tight junction proteins as markers of the blood–testis barrier. Long day (LD) photoperiod (16L : 8D) adult hamsters were exposed to short day (SD) photoperiod (8L : 16D) for 11 weeks to suppress gonadotrophins and then received FSH for up to 10 days. Sertoli cell proliferation was assessed immunohistochemically by the colocalisation of GATA-4 and PCNA, and quantified by stereology. Tight junction proteins (occludin and ZO-1) were colocalised using confocal microscopy. Sertoli cell proliferation in both the LD and SD controls was minimal; however, in response to FSH treatment proliferation was upregulated within 4 days compared with SD controls (98% v. 2%, P < 0.001, respectively). Tight junction proteins colocalised at the blood–testis barrier in LD hamsters, but were disorganised within the Sertoli cell cytoplasm in SD animals. FSH treatment restores colocalisation in a time-dependent manner. It is concluded that FSH contributes to the regulation of Sertoli cell proliferation and tight junction formation in the adult Djungarian hamster. This data provides definitive evidence that the adult Sertoli cell population in this model is modifiable by hormones. (1)Meachem et al. (2005). Biol Reprod 72, 1187.(2)Allan et al. (2004). Endocrinol 145, 1587.(3)Russell and Peterson (1985). Int Rev Cytol 94, 177.

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FSH and testosterone signaling in Sertoli cells

Reproduction ◽

10.1530/rep.1.00358 ◽

2005 ◽

Vol 130 (1) ◽

pp. 15-28 ◽

Cited By ~ 243

Author(s):

William H Walker ◽

Jing Cheng

Keyword(s):

Gene Expression ◽

Cell Proliferation ◽

Signaling Pathways ◽

Sertoli Cell ◽

Germ Cells ◽

Sertoli Cells ◽

Follicle Stimulating Hormone ◽

Reproductive Potential ◽

Recent Advances ◽

Stimulating Hormone

Testosterone and follicle-stimulating hormone (FSH) are required to obtain full reproductive potential. In the testis, somatic Sertoli cells transduce signals from testosterone and FSH into the production of factors that are required by germ cells as they mature into spermatozoa. Recent advances in identifying new signaling pathways that are regulated by FSH and testosterone have allowed for refinement in the understanding of the independent, overlapping and synergistic actions of these hormones. In this review, we discuss the signaling pathways that are regulated by FSH and testosterone as well as the resulting metabolic and gene expression changes that occur as related to Sertoli cell proliferation, differentiation and the support of spermatogenesis.

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Molecular mechanisms underlying AMH elevation in hyperoestrogenic states in males

Scientific Reports ◽

10.1038/s41598-020-71675-7 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Clara Valeri ◽

María M. Lovaisa ◽

Chrystèle Racine ◽

Nadia Y. Edelsztein ◽

Marina Riggio ◽

...

Keyword(s):

Sertoli Cells ◽

Oestrogen Receptor ◽

Molecular Mechanisms ◽

Follicle Stimulating Hormone ◽

Androgen Insensitivity Syndrome ◽

Fetal Life ◽

Complete Androgen Insensitivity Syndrome ◽

Peutz Jeghers Syndrome ◽

Clinical Conditions ◽

Sertoli Cell Line

Abstract Anti-Müllerian hormone (AMH) is secreted by Sertoli cells of the testes from early fetal life until puberty, when it is downregulated by androgens. In conditions like complete androgen insensitivity syndrome (CAIS), AMH downregulation does not occur and AMH increases at puberty, due in part to follicle-stimulating hormone (FSH) effect. However, other conditions like Peutz-Jeghers syndrome (PJS), characterised by low FSH, also have increased AMH. Because both CAIS and PJS may present as hyperoestrogenic states, we tested the hypothesis that oestradiol (E2) upregulates AMH expression in peripubertal Sertoli cells and explored the molecular mechanisms potentially involved. The results showed that E2 is capable of inducing an upregulation of endogenous AMH and of the AMH promoter activity in the prepubertal Sertoli cell line SMAT1, signalling through ERα binding to a specific ERE sequence present on the hAMH promoter. A modest action was also mediated through the membrane oestrogen receptor GPER. Additionally, the existence of ERα expression in Sertoli cells in patients with CAIS was confirmed by immunohistochemistry. The evidence presented here provides biological plausibility to the hypothesis that testicular AMH production increases in clinical conditions in response to elevated oestrogen levels.

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Molecular mechanisms involved in Sertoli cell adaptation to glucose deprivation

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00235.2009 ◽

2009 ◽

Vol 297 (4) ◽

pp. E907-E914 ◽

Cited By ~ 50

Author(s):

María F. Riera ◽

María N. Galardo ◽

Eliana H. Pellizzari ◽

Silvina B. Meroni ◽

Selva B. Cigorraga

Keyword(s):

Germ Cell ◽

Glucose Uptake ◽

Sertoli Cell ◽

Sertoli Cells ◽

Molecular Mechanisms ◽

Lactate Concentration ◽

Glucose Deprivation ◽

Cell Development ◽

Germ Cell Development ◽

Glucose Levels

Sertoli cells provide the physical support and the necessary environment for germ cell development. Among the products secreted by Sertoli cells, lactate, the preferred energy substrate for spermatocytes and spermatids, is present. Considering the essential role of lactate on germ cell metabolism, it is supposed that Sertoli cells must ensure its production even in adverse conditions, such as those that would result from a decrease in glucose levels in the extracellular milieu. The aim of the present study was to investigate 1) a possible effect of glucose deprivation on glucose uptake and on the expression of glucose transporters in rat Sertoli cells and 2) the participation of different signal transduction pathways in the above-mentioned regulation. Results obtained show that decreasing glucose levels in Sertoli cell culture medium provokes 1) an increase in glucose uptake accompanied by only a slight decrease in lactate production, 2) an increase in GLUT1 and a decrease in GLUT3 expression, and 3) an activation of AMP-activated protein kinase (AMPK)-, phosphatidylinositol 3-kinase (PI3K)/PKB-, and p38 MAPK-dependent pathways. Additionally, by using specific inhibitors of these pathways, a possible participation of AMPK- and p38MAPK-dependent pathways in the regulation of glucose uptake and GLUT1 expression is shown. These results suggest that Sertoli cells adapt to conditions of glucose deprivation to ensure an adequate lactate concentration in the microenvironment where germ cell development occurs.

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In Vitro Sertoli Cell Bioassay of Follicle-Stimulating Hormone (FSH): Serum from Different Animal Species Alters the Morphology of Rat Sertoli Cells without Affecting Their Response to FSH

General and Comparative Endocrinology ◽

10.1006/gcen.1994.1106 ◽

1994 ◽

Vol 95 (1) ◽

pp. 99-108 ◽

Cited By ~ 1

Author(s):

Manuela Simoni ◽

Bettina Schuhmann ◽

Gerhard F. Weinbauer

Keyword(s):

Sertoli Cell ◽

Sertoli Cells ◽

Animal Species ◽

Follicle Stimulating Hormone ◽

Stimulating Hormone

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Establishment and applications of male germ cell and Sertoli cell lines

Reproduction ◽

10.1530/rep-15-0546 ◽

2016 ◽

Vol 152 (2) ◽

pp. R31-R40 ◽

Cited By ~ 16

Author(s):

Hong Wang ◽

Liping Wen ◽

Qingqing Yuan ◽

Min Sun ◽

Minghui Niu ◽

...

Keyword(s):

Cell Line ◽

Sertoli Cell ◽

Cell Lines ◽

Germ Cells ◽

Sertoli Cells ◽

Molecular Mechanisms ◽

Simian Virus 40 ◽

T Antigen ◽

Seminiferous Tubules ◽

Male Germ Cells

Within the seminiferous tubules there are two major cell types, namely male germ cells and Sertoli cells. Recent studies have demonstrated that male germ cells and Sertoli cells can have significant applications in treating male infertility and other diseases. However, primary male germ cells are hard to proliferatein vitroand the number of spermatogonial stem cells is scarce. Therefore, methods that promote the expansion of these cell populations are essential for their use from the bench to the bed side. Notably, a number of cell lines for rodent spermatogonia, spermatocytes and Sertoli cells have been developed, and significantly we have successfully established a human spermatogonial stem cell line with an unlimited proliferation potential and no tumor formation. This newly developed cell line could provide an abundant source of cells for uncovering molecular mechanisms underlying human spermatogenesis and for their utilization in the field of reproductive and regenerative medicine. In this review, we discuss the methods for establishing spermatogonial, spermatocyte and Sertoli cell lines using various kinds of approaches, including spontaneity, transgenic animals with oncogenes, simian virus 40 (SV40) large T antigen, the gene coding for a temperature-sensitive mutant ofp53, telomerase reverse gene (Tert), and the specific promoter-based selection strategy. We further highlight the essential applications of these cell lines in basic research and translation medicine.

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