A Mouse Model for Imprinting of the Human Retinoblastoma Gene

Tasiou, Vasiliki; Hiber, Michaela; Steenpaß, Laura

doi:10.1371/journal.pone.0134672

Cited by 5 publications

(4 citation statements)

References 57 publications

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“…After reaching 50 to 60% confluence, cells were incubated with 0.1 µg/mL colcemid (Sigma-Aldrich, St. Louis, MO, USA) for 3 h [21]. Afterwards they were washed with phosphate buffer (PBS, Lonza, Basel, Switzerland) before the evaluation of the proliferation.…”

Section: Methodsmentioning

confidence: 99%

Effects of Insulin on Porcine Neonatal Sertoli Cell Responsiveness to FSH In Vitro

Cannarella

Arato

Condorelli

et al. 2019

JCM

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There is ongoing debate as to whether the decline of sperm production in recent times may be related to a parallel increase in the rate of obesity and diabetes. Lower anti-Müllerian hormone (AMH) and inhibin B secretion have been observed in young hyperinsulinemic patients compared to healthy controls, suggesting a Sertoli cell (SC) dysfunction. The pathophysiological mechanisms underlying SC dysfunction in these patients are poorly understood. To the best of our knowledge, no evidence is available on the effects of insulin on SC function. Therefore, this study was undertaken to assess the effects of insulin on basal and follicle-stimulating hormone (FSH)-stimulated SC function in vitro. To accomplish this, we evaluated the expression of AMH, inhibin B and FSHR genes, the secretion of AMH and inhibin B and the phosphorylation of AKT473 and SC proliferation on neonatal porcine SC after incubation with FSH and/or insulin. We found that similar to FSH, the expression and secretion of AMH is suppressed by insulin. Co-incubation with FSH and insulin decreased AMH secretion significantly more than with FSH alone. Insulin had no effect on the expression and secretion of the inhibin B gene, but co-incubation with FSH and insulin had a lower effect on inhibin B secretion than that found with FSH alone. FSH and/or insulin increased AKT473 phosphorylation and SC proliferation. In conclusion, the results of this study showed that insulin modulates SC function. We hypothesize that hyperinsulinemia may therefore influence testicular function even before puberty begins. Therefore, particular care should be taken to avoid the onset of hyperinsulinemia in children to prevent a future deleterious effect on fertility.

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Section: Methodsmentioning

confidence: 99%

Effects of Insulin on Porcine Neonatal Sertoli Cell Responsiveness to FSH In Vitro

Cannarella

Arato

Condorelli

et al. 2019

JCM

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“…After reaching 50-60% confluence, cells were treated with 0.1 μg/mL colcemid (Sigma-Aldrich, St. Louis, MO, USA) for 3 hours in the incubator ( 55 ) and washed with phosphate buffer (PBS). For the cell proliferation assay, SCs were incubated with 1 μM 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE, Sigma-Aldrich, St. Louis, MO, USA) in PBS for 8 min and washed with HBSS medium three times.…”

Section: Methodsmentioning

confidence: 99%

Sperm-carried IGF2 downregulated the expression of mitogens produced by Sertoli cells: A paracrine mechanism for regulating spermatogenesis?

et al. 2022

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IntroductionInsulin-like growth factor 2 (IGF2) mRNA has been found in human and mouse spermatozoa. It is currently unknown whether the IGF2 protein is expressed in human spermatozoa and, if so, its possible role in the cross-talk between germ and Sertoli cells (SCs) during spermatogenesis.MethodsTo accomplish this, we analyzed sperm samples from four consecutive Caucasian men. Furthermore, to understand its role during the spermatogenetic process, porcine SCs were incubated with increasing concentrations (0.33, 3.33, and 10 ng/mL) of recombinant human IGF2 (rhIGF2) for 48 hours. Subsequently, the experiments were repeated by pre-incubating SCs with the non-competitive insulin-like growth factor 1 receptor (IGF1R) inhibitor NVP-AEW541. The following outcomes were evaluated: 1) Gene expression of the glial cell-line derived neurotrophic factor (GDNF), fibroblast growth factor 2 (FGF2), and stem cell factor (SCF) mitogens; 2) gene and protein expression of follicle-stimulating hormone receptor (FSHR), anti-Müllerian hormone (AMH), and inhibin B; 3) SC proliferation.ResultsWe found that the IGF2 protein was present in each of the sperm samples. IGF2 appeared as a cytoplasmic protein localized in the equatorial and post-acrosomal segment and with a varying degree of expression in each cell. In SCs, IGF2 significantly downregulated GDNF gene expression in a concentration-dependent manner. FGF2 and SCF were downregulated only by the highest concentration of IGF2. Similarly, IGF2 downregulated the FSHR gene and FSHR, AMH, and inhibin B protein expression. Finally, IGF2 significantly suppressed the SC proliferation rate. All these findings were reversed by pre-incubation with NVP-AEW541, suggesting an effect mediated by the interaction of IGF2 with the IGFR.ConclusionIn conclusion, sperm IGF2 seems to downregulate the expression of mitogens, which are known to be physiologically released by the SCs to promote gonocyte proliferation and spermatogonial fate adoption. These findings suggest the presence of paracrine regulatory mechanisms acting on the seminiferous epithelium during spermatogenesis, by which germ cells can influence the amount of mitogens released by the SCs, their sensitivity to FSH, and their rate of proliferation.

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“…After reaching 50%–60% confluence, cells were treated with 0.1 µg/mL colcemid (Sigma-Aldrich no. 10295892001) for 3 h in the incubator [13] and washed with phosphate buffer (PBS). For cell proliferation assay, SC were incubated with 1 µM 5(6)-carboxyfluorescein diacetate N-succinmidyl ester (CFSE, catalog no.…”

Section: Methodsmentioning

confidence: 99%

Effects of GH and IGF1 on Basal and FSH-Modulated Porcine Sertoli Cells In-Vitro

Cannarella

Mancuso

Condorelli

et al. 2019

JCM

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Several lines of evidence suggest that insulin-like growth factor 1 (IGF1) is involved in Sertoli cell (SC) proliferation and that its receptor (IGF1R) could mediate follicle-stimulating hormone (FSH) effects. To examine the role of the growth hormone (GH)-IGF1 axis on SC function, we evaluated the effects of GH and IGF1 on basal and FSH-modulated SC proliferation, as well as on anti-Müllerian hormone (AMH) and inhibin B expression and secretion in-vitro. SCs from neonatal pigs were incubated with (1) placebo, (2) 100 nM highly purified urofollitropin (hpFSH), (3) 100 nM recombinant GH (rGH), (4) 100 nM recombinant IGF1 (rIGF1), (5) 100 nM hpFSH plus 100 nM rGH, (6) 100 nM hpFSH plus 100 nM rIGF1, for 48 h. We found that IGF1, but not FSH nor GH, stimulated SC proliferation. Furthermore, an inhibitory effect of FSH, GH and IGF1 on AMH secretion, and a stimulatory role of FSH and IGF1, but not GH, on inhibin B secretion were found. These results suggest that the GH-IGF1 axis influences basal and FSH-modulated SC proliferation and function. We speculate that SC proliferation occurring in childhood might be supported by the increased serum IGF1 levels observed during this period of life.

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A Mouse Model for Imprinting of the Human Retinoblastoma Gene

Cited by 5 publications

References 57 publications

Effects of Insulin on Porcine Neonatal Sertoli Cell Responsiveness to FSH In Vitro

Effects of Insulin on Porcine Neonatal Sertoli Cell Responsiveness to FSH In Vitro

Sperm-carried IGF2 downregulated the expression of mitogens produced by Sertoli cells: A paracrine mechanism for regulating spermatogenesis?

Effects of GH and IGF1 on Basal and FSH-Modulated Porcine Sertoli Cells In-Vitro

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