Effect of Versican Protein on the Cumulus Expansion of Buffalo Oocytes Complexes and its Role Sperm Transport

The present study was designed aiming at modulating the cumulus expansion characteristics of poor quality buffalo oocytes in an attempt to improve their fertilization outcomes. Cumulus expansion is a known morphological indicator of successful maturation and subsequent development potential. It depends on extracellular matrix (ECM) assembly during in vitro maturation (mediated by versican) and its subsequent disassembly prior to fertilization (as a result of cleavage of versicans by ADAMTS-1). In this work, poor quality (BCB-) buffalo cumulus oocyte complexes (COCs) were cultured for in vitro maturation (IVM) in presence of Interleukin (IL – 1B) at 50 and 100ng/ml along with BCB(control). IL-1B is known to enhance the ADAMTS-1 expression and hence helps in ECM disassembly. Impact of IL-1B supplementation on cumulus expansion was assessed morphologically, by scanning electron microscopy (SEM) and by expression analysis of related genes. Further, numbers of zona bound sperm under altered conditions were also counted. Development rates of treated oocytes were also monitored vis-à-vis the control ones. Expression of different isoforms of versican as well as ADAMTS-1 was observed to be significantly different between cumulus cells of BCB+ and BCBCOCs. Upon IL1B supplementation, ADAMTS-1 expression increased in BCBCOCs along with corresponding cumulus expansion rates. SEM analysis also revealed improved cumulus expansion in IL-1B supplemented BCBCOCs. HAS2 and TNFAIP-6 were significantly up-regulated in IL-1B supplemented groups as compared to BCBcontrol oocytes while PTGS2 expression remained unaffected. Significantly more numbers of sperms crossed cumulus barrier, especially in 100ng/ml IL-1B group. Following in vitro fertilisation (IVF) and in vitro culture (IVC) of newly formed embryos, both cleavage and blastocyst development rates were improved with IL-1B addition as compared to BCBcontrol. From this work, we conclude that IL-1B supplementation in IVM medium can improve cumulus expansion and development ability of poor quality buffalo oocytes. Scholar H.O. D. Major Advisor Versican oocytes cumulus शोधार्थी मखु्य मार्ग दशग क विभार् गौरव कुमार चौबे डॉ. टी. के. दत्ता ए. बी. टी. सी orZeku v/;;u esa [kjkc xq.kork okys HkSal ds oocytes dh cumulus foLrkj fo‖ ks ’krkvksa esa vf/kfeJ.k djds mudh fu’ks pu ifj.kkeksa esa lq/kkj djus dh dks f‖ k‖ k fd;k x;kA cumulus foLrkj ,d lQy ifjiDork vkSj fodkl dh {kerk dk ,d Kkr :irked lqpd gSA ;g {kerk mldh matrix dh assembly (versican dh e/;LFkrk esa ) rFkk ADAMTS-1 }kjk VERSICAN dks rksMsa tkus ds ckn ij fuHkZj djrk gSA bl dke es [kjkc xq.kork okys oocytes(BCB) dks IL-1B ds mifLFkrh e IVM s ek/;e es BCB (control) ds lkFk Mky fn;sA IL-1 B dks IVM ek/;e es Mkyus ds ckn cumulus foLrkj gqvk tks fd SEM rFkk Cumulus foLrkj lEcfU/kr genes ds vk/kkj ij crk;k x;kA vkSj vkxs bl vf/kfeJ.k ds ckn “kqdzk.kqvks a dh l[a;k zona pellucida ds Åij fxuk x;kA versican ds fofHkUu isoforms rFkk lkFk gh ADAMTS-1 ds vfHkO;fDr Fkh BCB+ rFkk BCB esa cMs vUrj ds lkFk ns[kk x;kA IL1B, IVM ek/;e esa Mkyus ds ckn ADAMTS-1 dh vfHkO;fDr rFkk Cumulus foLrkj Hkh gqvk fo‖ ys’k.k Hkh bl foLrkj dh iqf’V djrk gS rFkk IL-1B Mkyus ds ckn HAS2 rFkk TNFAIP-6 dh Hkh vfHkO;fDr esa lq/kkj gqvk ysfdu PTGS 2 dh vfHkO;fDr ij [kklk vUrj ugha ns[kk x;kA xkSjrykc gs fd vf/kd ek=k esa “kqdzk.kqvks a dh l[a;k Hkh Cumulus barrier dh ikj djds fo‖ ks ’krj okys leqg esa zona pellucida ds Åij x;sA rFkk bl vf/kfeJ.k ds ckn cleavage vkSj blastocyst rate Hkh BCB (Control) dh rqyuk esa T;knk ik;ka rRi‖ pkr bl dke ls ;g fu’d’kZ fudyrk gS fd IL-1B , IVM ek/;e esa Mkyus ls [kjkc xq.kork okys HkSal ds oocytes ds fodkl djus dh {kkerk rFkk cumulus foLrkj esa lq/kkj fd;k tk ldrk gSaA शोधार्थी मखु्य अध्यक्ष मखु्य मार्ग दशग क