Effect of buffalo oviductal and follicular fluid proteins on in vitro sperm capacitation
2006
Kumaresan, A. | Ansari, M.R. | Garg, A.
The effect of oviductal and follicular fluid proteins was studied on capacitation of spermatozoa in buffaloes. oviducts and ovaries were collected from apparently healthy buffalo genital tracts (nonluteal and luteal stage of estrous cycle) and oviducts were separated into isthmus and ampulla. Each segment of oviduct (nonluteral and luteal) was flushed with PBS (pH 7.4). The flushing obtained were centrifuged(3000 rpm; 30min), filtered (0.2u) and frozen at 20 C Follicular fluid was aspiratedfrom the follicles (5 mm) of buffalo ovaries. Proteins in the oviductal and follicular fluid were precipitated, dialyzed, filtered (0.2u) and stored as lyophilized in cryovials at-20C. Six pooled good quality ejaculates from 2 Murrah buffalo bulls were used. Eash pooled ejaculate was divided into 6 parts and prior to freezing oviductal and follicular proteins were added @ 1 mg/ml of extended semen while ne part was frozen without any added proteins (control). Frozen thawed semen samples were evaluated for capacitation status during incubation at 37C for 4 h by inducing acrosome reaction with lysophosphotidyl choline. Results revealed that both oviductal and follicular fluid proteins induced a capacitation. The nonluteal isthmic proteins induced capacitation in significantly higher number of sperms when compared to other groups. Similarly, at 4 h there were significantly higher acrosome reacted spermatozoa in follicular fluid protein treated group than the control group. From this study, it may be inferred that oviductal and follicular fluid proteins induced capacitation in higher percentage of sperm when incubated for 4 h at 37C and among the oviductal proteins, the monluteal isthmic proteins induced capacitation in significantly higher number of sperms, which may be the site for inducing capacitation of inseminated spermatozoa at estrus.
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