Online ISSN : 2349-8080 Issues : 12 per year Publisher : Excellent Publishers Email : editorinchiefijcrbp@gmail.com |
2Senior Research Fellow, ABC Division, National Dairy Research Institute, Karnal, Haryana, India
3Principal Scientist, ABC Division, National Dairy Research Institute, Karnal, Haryana, India
The study was designed to investigate the effect of plant based soya milk Tris (SMT)- extender used for cryopreservation of buffalo (Bubalus bubalis) spermatozoa on the expression of Bcl-2 family proteins that regulates the susceptibility of cells to apoptosis. Fresh buffalo semen (control without dilution) was cryopreserved in conventional EYT (20% egg yolk v/v in Tris) and SMT (25% soya milk v/v in Tris) extender and used for the assessment of expression of Bcl-2 family proteins. Proteins extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random were separated using SDS–PAGE followed by immunoblotting against Bcl-2, Bcl-xl, Bax and Bak. In addition, comet assay was performed to compare sperm DNA integrity in frozen thawed semen samples. The obtained results indicated a significant (p < 0.05) reduction in the expression of Bcl-2, Bax and Bak in cryopreserved semen samples. The results clearly demonstrated a significant (p<0.05) higher expression of Bcl-2 protein in SMT extender as compared to EYT extender indicating its better resistance to cryodamage.