Interactions of Sertoli Cells with Myoid Cells in vitro

We have examined the adhesion of primary Sertoli cells to a seminiferous tubule basement membrane (STBM) preparation in vitro. The STBM isolation procedure (Watanabe, T.K., L.J. Hansen, N.K. Reddy, Y.S. Kanwar, and J.K. Reddy, 1984, Cancer Res., 44:5361-5368) yields segments of STBM that retain their histotypic form in both three-dimensional tubular geometry and ultrastructural appearance. The STBM sleeves contain two laminae: a thick, inner basal lamina that was formed in vivo between Sertoli cells and peritubular myoid cells; and a thinner, outer basal lamina that was formed between myoid cells and sinusoidal endothelial cells. Characterization by immunofluorescence and SDS PAGE revealed that the isolated STBM retained fibronectin, laminin, and putative type IV collagen among its many components. When the STBM sleeves were gently shaken with an enriched fraction of primary Sertoli cells, the Sertoli cells bound preferentially to the lumenal basal lamina at the ends of the STBM sleeves. Few Sertoli cells bound to either the outer basal lamina of the STBM sleeves or to vascular extracellular matrix material which contaminated the STBM preparation. 3T3 cells, in contrast, bound to all surfaces of the STBM sleeves. Pretreatment of the STBM sleeves with proteases, 0.1 M Na metaperiodate, 4 M guanidine HCl, or heating to 80 degrees-90 degrees C inhibited lumenal Sertoli cell binding, but binding was not inhibited by chondroitinase ABC, heparinase, hyaluronidase, or 4 M NaCl. The lumenal Sertoli cell binding occurred in the presence or absence of added soluble laminin, but not fibronectin. The addition of soluble laminin, but not fibronectin, restored random binding of Sertoli cells to trypsinized STBM sleeves. Our in vitro model system indicates that Sertoli cells recognize differences in two basal laminae produced in vivo on either side of myoid cells.

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Follicle-stimulating hormone-dependent estrogen secretion by rat Sertoli cells in vitro: Modulation by calcium

Acta Endocrinologica ◽

10.1530/acta.0.1250280 ◽

1991 ◽

Vol 125 (3) ◽

pp. 280-285 ◽

Cited By ~ 6

Author(s):

J. Alan Talbot ◽

Ann Lambert ◽

Robert Mitchell ◽

Marek Grabinski ◽

David C. Anderson ◽

...

Keyword(s):

Sertoli Cells ◽

Culture Medium ◽

Follicle Stimulating Hormone ◽

Dibutyryl Camp ◽

Immature Rat ◽

Estradiol Secretion ◽

Old Rats ◽

Stimulating Hormone

Abstract We have investigated the role of Ca2+ in the control of FSH-induced estradiol secretion by Sertoli cells isolated from 8-10 days old rats. Exogenous Ca2+ (4-8 mmol/1) inhibited FSH-stimulated E2 secretion such that, with 8 mmol/l Ca2+ and FSH (8 IU/l) E2 secretion decreased from 2091±322 to 1480±84 pmol/l (p<0.002), whilst chelation of Ca2+ in the culture medium with EGTA (3 mmol/l) increased E2 secretion from 360±45 to 1242±133 pmol/l) in the absence of FSH. Further, EGTA (3 mmol/l) markedly potentiated FSH (8 IU/l), forskolin (1 μmol/l) and dibutyryl cAMP (1 mmol/l)-stimulated E2 secretion. Addition of the Ca2+ ionophores, ionomycin (2-5 μmol/l) and A23187 (2 μmol/l), inhibited FSH (8 IU/l)-stimulated E2 secretion by >80%. The effect of ionomycin was totally reversible, whereas that of A23187 was irreversible. Ionomycin (5 μmol/l) had no effect on EGTA-induced E2 secretion in the absence of FSH, but reduced EGTA-provoked E2 secretion by 59% in the presence of FSH (8 IU/l). Similarly, forskolin- and dibutyryl cAMP-provoked E2 production was inhibited 46-50% by ionomycin (5 μmol/l). We conclude that FSH-induced E2 secretion from immature rat Sertoli cells is modulated by intra- and extracellular Ca2+.

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TCF21+ mesenchymal cells contribute to testis somatic cell development, homeostasis, and regeneration in mice

Nature Communications ◽

10.1038/s41467-021-24130-8 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Yu-chi Shen ◽

Adrienne Niederriter Shami ◽

Lindsay Moritz ◽

Hailey Larose ◽

Gabriel L. Manske ◽

...

Keyword(s):

Tissue Homeostasis ◽

Testicular Development ◽

Myoid Cells ◽

Regenerative Capacity ◽

Adult Testis ◽

Mesenchymal Progenitors ◽

And Function ◽

Potential Reserve ◽

Resident Fibroblast

AbstractTesticular development and function rely on interactions between somatic cells and the germline, but similar to other organs, regenerative capacity declines in aging and disease. Whether the adult testis maintains a reserve progenitor population remains uncertain. Here, we characterize a recently identified mouse testis interstitial population expressing the transcription factor Tcf21. We found that TCF21lin cells are bipotential somatic progenitors present in fetal testis and ovary, maintain adult testis homeostasis during aging, and act as potential reserve somatic progenitors following injury. In vitro, TCF21lin cells are multipotent mesenchymal progenitors which form multiple somatic lineages including Leydig and myoid cells. Additionally, TCF21+ cells resemble resident fibroblast populations reported in other organs having roles in tissue homeostasis, fibrosis, and regeneration. Our findings reveal that the testis, like other organs, maintains multipotent mesenchymal progenitors that can be potentially leveraged in development of future therapies for hypoandrogenism and/or infertility.

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Ezrin is an Actin Binding Protein That Regulates Sertoli Cell and Spermatid Adhesion During Spermatogenesis

Endocrinology ◽

10.1210/en.2014-1163 ◽

2014 ◽

Vol 155 (10) ◽

pp. 3981-3995 ◽

Cited By ~ 19

Author(s):

N. Ece Gungor-Ordueri ◽

Elizabeth I. Tang ◽

Ciler Celik-Ozenci ◽

C. Yan Cheng

Keyword(s):

Sertoli Cell ◽

Sertoli Cells ◽

Adherens Junction ◽

Actin Binding ◽

Permeability Barrier ◽

Tight Junction Permeability ◽

Residual Bodies ◽

Cell Interface

Abstract During spermatogenesis, the transport of spermatids and the release of sperms at spermiation and the remodeling of the blood-testis barrier (BTB) in the seminiferous epithelium of rat testes require rapid reorganization of the actin-based cytoskeleton. However, the mechanism(s) and the regulatory molecule(s) remain unexplored. Herein we report findings that unfold the functional significance of ezrin in the organization of the testis-specific adherens junction at the spermatid-Sertoli cell interface called apical ectoplasmic specialization (ES) in the adluminal compartment and the Sertoli cell-cell interface known as basal ES at the BTB. Ezrin is expressed at the basal ES/BTB in all stages, except from late VIII to IX, of the epithelial cycle. Its knockdown by RNA interference (RNAi) in vitro perturbs the Sertoli cell tight junction-permeability barrier via a disruption of the actin microfilaments in Sertoli cells, which in turn impeded basal ES protein (eg, N-cadherin) distribution, perturbing the BTB function. These findings were confirmed by a knockdown study in vivo. However, the expression of ezrin at the apical ES is restricted to stage VIII of the cycle and limited only between step 19 spermatids and Sertoli cells. A knockdown of ezrin in vivo by RNAi was found to impede spermatid transport, causing defects in spermiation in which spermatids were embedded deep inside the epithelium, and associated with a loss of spermatid polarity. Also, ezrin was associated with residual bodies and phagosomes, and its knockdown by RNAi in the testis also impeded the transport of residual bodies/phagosomes from the apical to the basal compartment. In summary, ezrin is involved in regulating actin microfilament organization at the ES in rat testes.

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Thyroid hormone modulates androgen and oestrogen receptor content in the Sertoli cells of peripubertal rats

Journal of Endocrinology ◽

10.1677/joe.0.1480043 ◽

1996 ◽

Vol 148 (1) ◽

pp. 43-50 ◽

Cited By ~ 26

Author(s):

M L Panno ◽

D Sisci ◽

M Salerno ◽

M Lanzino ◽

V Pezzi ◽

...

Keyword(s):

Sertoli Cells ◽

Oestrogen Receptor ◽

Age Groups ◽

Strong Relationship ◽

Inhibitory Influence ◽

Aromatase Activity ◽

Severe Impairment ◽

Cellular Compartments

Abstract A possible role of tri-iodothyronine (T3) on the interplay between testicular steroids and Sertoli cells has been investigated on the basis of previous findings demonstrating a direct inhibitory influence of T3 on aromatase activity and oestradiol production in peripuberal Sertoli cells. In this context, the present study was focused on the effects of T3 on oestrogen receptor (ER) and androgen receptor (AR) contents in the cytosol and nucleus of Sertoli cells isolated from 2-, 3- and 4-week-old euthyroid, hypothyroid and hypothyroid treated rats. Hypothyroidism was induced by the oral administration of 0·025% methimazole (MMI) from birth until the rats were killed at 2, 3 and 4 weeks of age. Half of the MMI-treated animals were injected i.p. with l-tri-iodothyronine (T3; 3 μg/100 g body weight) during the last week before death. Sertoli cells from all groups were initially cultured under basal conditions for the first 24 h and subsequently in the presence of testosterone with or without T3 for an additional 24 h. Hypothyroidism was associated with severe impairment of body as well as testicular growth. Euthyroid ERs showed an elevated Kd (0·76 nm) which was similar in the different age groups investigated. The in vitro addition of T3 or testosterone induced a decrease in ER content and this decrease was greater after exposure to both hormones. In 2- and 3-week-old hypothyroid rats, ER content was markedly increased and was reversed in euthyroid rats when T3 was given in vivo. When ERs were assayed in the Sertoli cell nucleus and cytoplasm of 2- and 3-week-old animals, a strong relationship in ER content in the two cellular compartments was observed. Neither of the hormones tested seemed to affect the AR content in the nucleus significantly, while the in vitro addition of testosterone or T3 or both hormones together augmented the ARs in the cytosol to a greater extent, resulting in an increase in their total (cytosolic and nuclear) content in the cells. The present data suggest that T3 down-regulates ERs and up-regulates ARs in peripuberal Sertoli cells. The additive effect of testosterone and T3 in up-regulating ARs could possibly involve a role for T3 in influencing the androgen responsiveness of the Sertoli cells during spermatogenesis. Journal of Endocrinology (1996) 148, 43–50

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The enhancement of neural stem cell survival and growth by coculturing with expanded sertoli cells in vitro

Biotechnology Progress ◽

10.1002/btpr.720 ◽

2011 ◽

Vol 28 (1) ◽

pp. 196-205 ◽

Cited By ~ 6

Author(s):

Bingyang Shi ◽

Lei Deng ◽

Xiaolin Shi ◽

Sheng Dai ◽

Hu Zhang ◽

...

Keyword(s):

Stem Cell ◽

Cell Survival ◽

Neural Stem Cell ◽

Sertoli Cells ◽

Stem Cell Survival ◽

Survival And Growth

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Reproductive toxicity of heavy metals in fallow deer in vitro

Acta Veterinaria Brno ◽

10.2754/avb202190030277 ◽

2021 ◽

Vol 90 (3) ◽

pp. 277-286

Author(s):

Ehdaa Eltayeb Eltigani Abdelsalam ◽

Hana Banďouchová ◽

Tomáš Heger ◽

Miroslava Kaňová ◽

Kateřina Kobelková ◽

...

Keyword(s):

Heavy Metals ◽

Sertoli Cells ◽

Reproductive Toxicity ◽

Toxic Metal ◽

Fallow Deer ◽

Ovarian Follicles ◽

Metal Exposure ◽

Lipid Hydroperoxides ◽

Single Strand Break

Sertoli cells play a crucial role in male fertility through boosting and regulating the differentiation of spermatogonial stem cells into mature sperm during spermatogenesis. Female ovarian follicles are responsible for the production of mature ova and control of ovarian steroidogenesis. Disruption of these structures through exposure to environmental pollutants is critical for reproductive health. Here, we derived primary cell cultures of Sertoli cells and ovarian follicles from fallow deer (Dama dama). Cells were used as in vitro models to explore reproductive toxicity of heavy metals in wild species. Adverse effects of cadmium (CdCl2), methylmercury (MeHgCl2), and lead (PbCl2) were investigated through a range of equal molar concentrations (0, 15, 30, 60, 125, 250 µM). We found both concentration-dependent and independent cytotoxic patterns (P < 0.01, P < 0.05) in cells exposed to CdCl2, MeHgCl2, and PbCl2. Based on generation of lipid hydroperoxides, significant levels of cell oxidative perturbation were detected in the CdCl2 (P = 0.0001), PbCl2 (P = 0.001), and MeHgCl2 (P = 0.003) groups. Likewise, the antioxidant enzymes catalase and glutathione peroxidase were inhibited in all metal-treated groups (P < 0.01). Genotoxic DNA damage (single-strand break) was also observed (MeHgCl2 group, P = 0.002; CdCl2 and PbCl2 groups, P = 0.004). Increased activity of superoxide dismutase (P = 0.0002 and P = 0.01) was observed in MeHgCl2 and CdCl2, respectively. Cell apoptosis was detected in all the PbCl2 and CdCl2 (P = 0.00007) and MeHgCl2 (P = 0.001) groups. The results of this study can be used to characterize the responsiveness of fallow deer gonadal cells to the stress of toxic metal exposure.

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Regulation of GDNF expression in Sertoli cells

Reproduction ◽

10.1530/rep-18-0239 ◽

2019 ◽

Cited By ~ 8

Author(s):

Parag Parekh ◽

Thomas Xavier Garcia ◽

Marie-claude Hofmann

Keyword(s):

Germ Cell ◽

Sertoli Cells ◽

Seminiferous Tubules ◽

Receptor Complex ◽

Myoid Cells ◽

Self Renewal ◽

Cell Proliferation And Differentiation ◽

Proliferation And Differentiation ◽

Direct Cell ◽

Ability To Drive

Sertoli cells regulate male germ cell proliferation and differentiation and are a critical component of the spermatogonial stem cell (SSC) niche, where homeostasis is maintained by the interplay of several signaling pathways and growth factors. These factors are secreted by Sertoli cells located within the seminiferous epithelium, and by interstitial cells residing between the seminiferous tubules. Sertoli cells and peritubular myoid cells produce glial cell line-derived neurotrophic factor (GDNF), which binds to the RET/GFRA1 receptor complex at the surface of undifferentiated spermatogonia. GDNF is known for its ability to drive SSC self-renewal and proliferation of their direct cell progeny. Even though the effects of GDNF are well studied, our understanding of the regulation its expression is still limited. The purpose of this review is to discuss how GDNF expression in Sertoli cells is modulated within the niche, and how these mechanisms impact germ cell homeostasis.

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