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ThesisItem Open Access GENE EXPRESSION PATTERNS OF SELECTIVE AQUAPORINS IN BUBALINE SPERMATOZOA(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-07) UDAY SIVA NAGA SANKAR, A; SRINIVASA PRASAD, CH(MAJOR); IQBAL HYDER; MUTHA RAO, M; ASWANI KUMAR, KThe present study was designed to assess the gene expression patterns of specific aquaporins (AQP’s) in bubaline spermatozoa. Eight healthy Murrah bulls aged between three to six years were selected from ABC Semen Station, Veeravalli, Krishna District, Andhra Pradesh. Fresh and frozen semen samples were collected for the study. As a preliminary quality evaluation, fresh semen samples (4 samplings from each bull, n=32) were examined for semen volume, sperm concentration, mass motility, individual progressive motility, abnormalities, viability, hypo-osmotic swelling test (HOST) and acrosomal integrity. The corresponding mean±SE values were found to be 4.337±0.29mL, 1147±3.10million/mL, 3.637±0.14, 71.5±0.18%, 9.36±0.82, 74.42±0.49%, 74.92±0.56% and 86.40±0.70%, respectively. Similarly, frozen semen samples (four straws per sample, n=32) were also evaluated for concentration, post thaw motility, viability, HOST and acrosomal integrity and the mean±SE values were found to be 19.68±0.28million/straw, 51.82±0.23%, 62.15±0.45%, 51.10±1.17% and 74.12±1.043%, respectively. The functional parameters were compared between fresh and frozen samples using unpaired t-test and it was observed that percent viability, HOST and acrosomal integrity were significantly (p0.05) difference was noticed between the down regulated groups. The expression of AQP7 showed no significant (p>0.05) difference between control and frozen non capacitated group, while the expression was significantly (p<0.05) upregulated in frozen capacitated compared to fresh capacitated group. The mRNA expression of AQP8 was significantly (p<0.05) down-regulated in fresh capacitated and frozen non-capacitated groups compared to fresh non-capacitated control. Further, a significant (p<0.05) increase in the expression of AQP8 was noticed in frozen capacitated compared to fresh capacitated group. The mRNA expression of AQP11 was significantly (p<0.05) upregulated in frozen capacitated compared to fresh capacitated and frozen non capacitated groups. The expression pattern of AQP3 is indicative of intense oxidative stress during capacitation and cryopreservation, while the expression pattern of aquaglyceroporins AQP7 and 11 is suggestive of its protective role in cryopreservation (AQP7) and freeze thaw process (AQP7 and 11) via influx and efflux of glycerol which was added to tris-yolk citrate extender. Similarly, AQP8 being a peroxiporin, its protective role in combating oxidative damage by excess ROS formed due to cumulative effect of both cryopreservation and capacitation has been noticed. The present study confirmed the presence of AQP3, 7, 8 and 11 in Murrah buffalo spermatozoa. Further, the study revealed that the studied genes have potential role in cryotolerance (AQP7), capacitation induced oxidative stress (AQP8) and freeze thaw process (AQP 7 and 11) of frozen semen.
