In vitro induction of catfish, Clarias batrachus, oocyte maturation by conspecific vitellogenin 1 (CFVg1).

Abstract

Present study demonstrates that conspecific vitellogenin1 (CFVg1) induces oocyte maturation in the catfish, Clarias batrachus. CFVg1 is able to develop fertilizable eggs in the Clarias batrachus. Therefore, different in vitro oocyte culture experiments were designed to see whether CFVg1 has efficacy of oocyte maturation and its pathway. In in vitro oocyte culture experiment, CFVg1 showed a dose- and time-dependent response and 64% maturation was obtained at the dose level of 10µg/ml or more. CFVg1 induction of oocyte maturation was confirmed by co-incubating CFVg1 with CFVg1-antiserum (a-CFVg1), which inhibited the CFVg1-induced oocyte maturation. To answer issues lead to the understanding of the mechanism of vitellogenin (Vg) on oocyte maturation, trypsin digested CFVg1 and Indian major carp Cirhinus mrigala Vg HAI (Hydroxy appetite peak I) also showed significant level of maturation. Actinomycin-D and cycloheximide blocked the effect of CFVg1, indicating that CFVg1 acts through transcription and translation. Theophylline, the phosphodiesterase inhibitor, and cAMP also inhibited the stimulatory effect of CFVg1 on oocyte maturation, indicating indirectly that CFVg1-induced oocyte maturation by decreasing the intracellular cAMP possibly by activating the phosphodiesterase enzyme. Trilostane, the 3β-HSD-blocker, did not inhibit the CFVg1-induced oocyte maturation but wortmannin and Ly294002 two mechanistically different specific inhibitors of PI3 kinase blocked the oocyte maturation. The results thus indicate that oocyte maturation in catfish by Vg may be regulated by two pathways: (1) through decreasing the intraoocyte cAMP level by activating the cAMP-PKA pathway and (2) by cAMP-dependent PI3K/Akt pathway. Therefore, there might be role of vitellogenin itself in initiation of oocyte maturation.