Singh, VijayShah, Nadeem2017-11-102017-11-102016-07-27http://krishikosh.egranth.ac.in/handle/1/5810035331Effect Of Addition Of Glutathione On Tyrosine Phosphorlation And Apoptosis Like Changes In Cryopreserved Hariana Bull SemenThe present study evaluated the beneficial effects of incorporation of Glutathione as an additive in Tris egg yolk based extender in Hariana bull semen opted for ultralow freezing. The study valuated physical seminal attributes (motility, livability and membrane integrity), process of ryocapacitation, immunoblotting for identification of tyrosine phosphorylated proteins, immunolocalization of osine phosphoproteins in spermatozoa, and apoptosis like changes in terms of mitochondrial transmembrane potential and DNA integrity during after equilibration and thawing. Ten ejaculates from four Hariana bulls were divided into three aliquots: One aliquot diluted with egg yolk tris citrate (EYTC) extender (Control), second aliquot was diluted with EYTC but supplemented with 0.5Mm Glutathione (T1) and the third aliquot was diluted with EYTC and supplemented with 1.0Mm Glutathione (T2) and were cryopreserved. Semen evaluation at equilibration and post-thaw showed supplementation of Glutathione (0.5Mm) to EYTC extender significantly (P < 0.05) increased motility, viability and membrane integrity of spermatozoa. The degree of cryocapacitation was significantly (P < 0.05) decreased in Glutathione supplemented group. Immunoblot revealed six proteins which were tyrosine phosphorylated and protein of 30kDa (p30) showed differential variation in intensity in the three samples. There was significant reduction in band intensity of 30kDa in T1 as compared to control and T2. It was also found that tyrosine phosphorylated proteins were differently located during different stages of semen preservation. Addition of GSH significantly decreased percentage of spermatozoa showing fragmented DNA after thawing as compared to control. Along with this, GSH supplementation significantly increased the percentage of spermatozoa with high transmembrane mitochondrial potential. The result of the present study clearly demonstrated beneficial effects of Glutathione supplementation on post thaw cryocapacitation and apoptosis like changes in spermatozoa and it can be suitably incorporated for long term preservation of spermatozoa.ennullThesis