Effects of IGF-1, TGF-alpha plus TGF-beta1 and bFGF on in vitro survival, growth and apoptosis in FSH-stimulated buffalo (Bubalis bubalus) preantral follicles.
ABSTRACT Investigate the effect of various growth factors viz. IGF-I, TGF-alpha + TGF-beta1 and bFGF either alone or in combination, with FSH on in vitro growth, survival, antrum formation, steroidogenesis and apoptosis of buffalo preantral follicles (PFs).
Buffalo ovaries were collected from abattoir; PFs were isolated and divided into five treatment groups. TCM-199 supplemented with 10% FBS, 1% ITS+EGF+FSH control (group a), control+IGF-I (group b), control + TGF-alpha + TGF-beta1 (group c), control + IGF-I + TGF-alpha + TGF-beta1 (group d) and control+bFGF (group e). Progesterone (P4) and 17beta-estradiol (E2) concentrations were evaluated by RIA and apoptosis by TUNEL assay.
TGF-alpha + TGF-beta1 inhibited follicular survival and induced oocyte apoptosis, while IGF-I + TGF-alpha + TGF-beta1 suppressed this inhibitory action. IGF-I significantly (P < 0.05) enhanced the follicle survival, growth and induced antrum formation. FGF had greater effects on both survival and growth rate of oocytes than other treatment groups. Progesterone and estradiol accumulation was significantly (P < 0.05) greater in presence of FGF and IGF-I than TGF-alpha + TGF-beta1.
Survival, growth, antrum formation and steroidogenesis are stimulated by IGF-I and bFGF, whereas TGF-alpha + TGF-beta1 inhibited growth and survival of PFs which led to induced oocyte apoptosis in buffalo PFs.
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ABSTRACT: Cryopreservation of ovarian tissue or follicles has been proposed as an alternative method for fertility preservation. Although successful vitrification of follicles has been reported in several mammalian species, the survival is generally low. The aim of this study was to investigate the effects of FGF and EGF on in vitro preantral follicle development after vitrification. Preantral follicles with diameter of 150-180 μm were mechanically isolated from 18-21 days old NMRI mice ovaries. Follicles were vitrified and warmed then cultured in ɑ- minimal essential medium supplemented with 20 ng/ml FGF, 20 ng/ml EGF and 20 ng/ml FGF +20 ng/ml EGF simultaneously. After 12 days hCG/EGF was added to culture medium and after 18-20 h later the presence of cumulus oocyte complexes (COC) and oocyte maturation were assessed. χ2 test was used to analyse survival and ovulation rate of the follicles. our results showed that the rate of MII oocytes in FGF group increased in comparison with control and treatment groups (P<0.027), but there was no difference between control and EGF and EGF+FGF groups in oocyte maturation rate (P>0.05). There was significant decrease in survival rate of follicles in EGF-FGE group in comparison with other groups (P<0.008). After in vitro ovulation induction the follicles in EGF group showed a higher ovulation rate (P<0.008) than those cultured in other groups. FGF has beneficial effect on oocyte maturation and EGF increases COCs number in vitro. Combination of EGF and FGE decrease the number of survived follicles.Cell Journal 10/2013; 16(3). · 0.46 Impact Factor
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ABSTRACT: The proliferating effect of basic fibroblast growth factor (bFGF) on granulosa cells from ovarian pre-hierarchical follicles was evaluated in the laying chickens. Expression of bFGF receptor 1 (FGFR1) from small yellow follicles was determined by immunohistochemistry and RT-PCR. The FGFR1 protein and mRNA were intensively expressed in the granulosa layer. After 8- to 24-h treatment with bFGF (0.1-100 ng/ml), the proliferation of cultured granulosa cells was remarkably enhanced in a dose- and time-dependent manner. The FGFR1 antagonist SU5402 inhibited bFGF-induced cell proliferation. This stimulating effect was further confirmed by 5-bromo-2-deoxyuridine incorporation and terminal transferase dUTP nick end-labelling assay. Immunocytochemistry of protein kinases A (PKA) and C (PKC) showed that the pro-proliferation action of bFGF predominantly activated PKC expression. Meanwhile, the bFGF-induced cell proliferation was significantly promoted by PKC activator PMA and inhibited by PKC inhibitor H(7) (p < 0.05). In addition, the bFGF-elicited cell proliferation was accompanied with increased mRNA expression of the cell cycle-regulating genes including cyclins D1 and E1, cyclin-dependent kinases 2 and 6. In conclusion, bFGF promoted the proliferation of ovarian granulosa cells through binding with FGFR1 and involving PKC pathway in the pre-hierarchical follicles of the laying chickens.Reproduction in Domestic Animals 05/2011; 47(1):135-42. DOI:10.1111/j.1439-0531.2011.01813.x · 1.18 Impact Factor
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ABSTRACT: The insulin-like growth factor-1 (IGF-1) plays an important role in the regulation of reproductive function. In the present study, we examined the effects of caloric restriction (CR) on the reproductive lifespan in rats and investigated the potential role of IGF-1. After 10 weeks of treatment, we determined the distribution of the ovarian follicles at various stages and measured the plasma level of IGF-1, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estrogen (ESG). Our results show that IGF-1 level was decreased after CR and correlated with the decrease in the levels of LH, FSH and ESG. Moreover, a higher percentage of primordial follicles and surviving follicles was observed in CR rats than in control rats (P<0.05). Immunohistochemical analysis showed that IGF-1 was extensively expressed in the cytoplasm of granulosa cells in the surviving follicles at different stages but not in the atretic follicles. Taken together, these results suggest that caloric restriction promotes the reproductive capacity of female rats via modulating the level of IGF-1, which then regulate pituitary gonadotrope cells to reduce the release of LH, FSH and ESG, and modulate follicular development.General and Comparative Endocrinology 09/2011; 174(2):232-7. DOI:10.1016/j.ygcen.2011.09.005 · 2.67 Impact Factor