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: This study aimed to evaluate the immunolocalization and messenger RNA (mRNA) expression for transforming growth factor-beta (TGF-β) and its receptors (TGF-βRI and RII), as well as mRNA expression for P450 aromatase and FSH receptor in caprine preantral follicles. The effects of TGF-β, FSH alone, or in association on the in vitro follicular development were also assessed. Immunohistochemical analyses showed the expression of TGF-β and its receptors in oocytes of all follicle stages and granulosa cells of primary and secondary follicles. mRNA for TGF-β receptors and for FSH receptor (FSHR) was present in preantral follicles as well as in oocytes and granulosa cells of antral follicles. Isolated secondary follicles were cultured in α-minimum essential medium (MEM) alone or supplemented with either FSH (100 ng/ml), TGF-β (10 ng/ml), or TGF-β + FSH for 18 d. TGF-β increased significantly oocyte diameter when compared to FSH alone and control. After 18 d of culture, all groups showed a significant reduction in P450 aromatase and FSHR mRNA levels in comparison to fresh control. In contrast, treatment with FSH significantly increased the mRNA expression for TGF-β in comparison to fresh control and other treatments. In conclusion, the findings showed that TGF-β and its receptors are present in caprine ovarian follicles. Furthermore, they showed a positive effect on oocyte growth in vitro.In Vitro Cellular & Developmental Biology - Animal 05/2014; · 1.29 Impact Factor
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ABSTRACT: It is known that caloric restriction extends lifespan and can minimize age-related dysfunction of the reproductive system. We became interested in how caloric restriction influences apoptosis, which is a crucial process that maintains ovarian cell homeostasis. We examined ovarian cells in: 2.5-year-old wild type mice on caloric restriction (CR) or fed ad libitum (AL) and Laron dwarf mice (GHR-KO) at the same ages on CR or fed AL. Apoptosis was assessed by histochemical analysis on paraffin sections of ovarian tissue. Morphological and histochemical analysis revealed that CR improved reproductive potential in 2.5-year-old WT littermates and GHR-KO female mice, as indicated by the increased number of ovarian follicles. The level of apoptosis in ovarian tissue was higher in WT mice on a CR diet compared with WT mice on the AL diet. In GHR-KO mice, the level of apoptosis in ovaries was similar for mice on CR and on AL diets and bigger than in WT mice on CR. Morphological and histochemical analysis revealed a younger biological age of the ovaries in 2-year-old WT littermates and GHR-KO female mice on CR compared with animals fed AL.Journal of Ovarian Research 09/2013; 6(1):67. · 2.43 Impact Factor
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ABSTRACT: To compare the effects of human menopausal gonadotropins (hMG) and recombinant follicle stimulating hormone (rFSH) on transforming growth factor (TGF) β1 concentration in the rat ovary. Twenty-one fertile Wistar-Albino rats were divided into 3 groups of 7. Groups 1, 2 and 3 were injected with saline, hMG or rFSH, respectively, over 5 days, after which they underwent ovariectomy. Hematoxylin and eosin (H&E) staining was used for histological examination. TGF β1 staining levels in ovarian stroma, vessel walls, granulosa cells of Graafian follicles and corpus luteum cells were investigated immunohistochemically. On histological examination, the number of smaller antral follicles was higher in the control group, while there were more and larger antral follicles in the hyperstimulated groups. There were statistically significant differences in staining in vessel walls and granulosa cells between the control and stimulated groups. Both stimulation protocols caused an increased TGF β1 concentration in vessel walls, while there was weak staining in granulosa cells in the treatment groups compared to the control group (p<0.05). There were no significant differences in staining scores between the two treatment groups (p>0.05). The effects of two different gonadotropin preparations on TGF β1 concentrations in different localizations in the rat ovaries are comparable. It may be postulated that the luteinizing hormone (LH) content of hMG contributes little or nothing to the TGF β1 mediated angiogenesis.European journal of obstetrics, gynecology, and reproductive biology 04/2012; 163(1):35-8. · 1.97 Impact Factor