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20111
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Author: Anand × End date: 2011 × Start date: 2011 × Type: Thesis ×
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    ThesisItemOpen Access
    Characterization of Growth and Atresia of Buffalo Ovarian Follicle by Biochemical Analysis and Expression Studies
    (Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar, 2011-10-15) Anand; Murthy, V.C.; Ravindra, J.P.; Jamuna, K.V.; Honnappa, T.G.; Selvaraju, S.
    The most important constraint of buffalo ovarian physiology that hampers reproductive efficiency is the availability of less number of primordial follicle, associated with high atretic rate as compared to cattle. Hence the objective of the present study was (i) to determine certain enzymatic and hormonal changes associated with growth and atresia of buffalo ovarian follicles and (ii) To identify the receptor in granulosa cells and the changes in the ovarian follicular fluid of buffalo. Buffalo ovarian follicular fluid was collected from slaughter house ovaries by aspiration and from live animals by transvaginal ultrasound guided follicular aspiration. Based on the Haematoxylin and Feulgen’s staining of granulosa cells nuclei follicles were classified into healthy and atretic (>5% of granulosa cells with pyknotic and fragmented nuclei were considered as atretic). Based on the surface diameter the follicles were classified in to small (3-5mm), medium (5-8mm) and large (>8mm) follicles. The study revealed that the presence of CL influenced the growth of the follicle in all sizes. The follicular fluid progesterone was significantly (P<0.05) high in ovulatory follicle. A significantly (P<0.05) higher estradiol-17β concentration has observed in medium sized non atretic follicle as compared atretic follicle. The presence of around 40-45 kDa (pI of 5-7) proteins in the medium size (5-8mm) atretic follicle, on other hand the presence of the 30 -35 kDa (pI of 6-8) proteins were observed in dominant ovulatory follicle. In live animals, the follicular fluid SOD concentration (IU/mg of protein) was significantly (P<0.05) higher in non-atretic follicles as compared to atretic follicles of 5-8 mm follicles. In live animals, the follicular fluid HDL cholesterol concentration in >8mm diameter atretic follicle was significantly (P<0.05) higher than non-atretic follicle. Expression studies revealed that the presence of LH receptor mRNA was detected only in healthy follicles of > 8 mm in diameter and the expression of LH receptor mRNA in granulosa cells was not detected in the regressing dominant follicles. The 3β HSD expression was significantly higher in non atretic follicles than atretic follicles. From This study it is evident that the medium size follicles with high estradiol- 17β concentration is responsible for dominance and the dominant follicle with higher progesterone concentration reaches ovulation. Study further revels that around 40-45 kDa (5-7 pI) proteins might be involved in the atresia of the medium size (5-8mm) follicle, where as 30 -35 kDa (6-8 pI) proteins might be involved in follicular dominance and ovulation. The present study showed that only the dominant follicle expressed LH receptor indicating lutenization and progressing toward ovulation.