GENE EXPRESSION PATTERNS OF SELECTIVE MONOCARBOXYLATE TRANSPORTERS IN BUBALINE SPERMATOZOA
Loading...
Files
Date
2023-10
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA
Abstract
The objective of the present study was to evaluate the gene expression of
certain monocarboxylate transporters in bubaline spermatozoa. Eight adult healthy
buffalo bulls between three to six years of age were randomly selected from ABC
Semen Station, Veeravalli, Krishna District, Andhra Pradesh. The fresh semen
samples were collected from eight buffalo bulls once in a week for four weeks by AV
method. As a preliminary quality check, fresh semen samples (4 samples from each
bull, n=32) were evaluated at the semen station for volume, concentration, mass
motility, individual progressive motility, viability, HOST, acrosomal integrity and the
mean±SE values were found to be 4.09±0.26mL, 1243.875±65.621 million/mL,
3.86±0.34 and 70.04±9.92%, 73.42±0.49%, 73.98±0.56% and 87.75±0.52%
respectively. With respect to evaluation of gene expression of monocarboxylate
transporters, the fresh semen samples were transported at 37°C. The samples were
divided in to two groups. Fresh uncapacitated as control (4 samples from each bull
n=32) and fresh capacitated (n=32). It was taken care that fresh semen samples had
sperm concentration of 50-60 million spermatozoa. In vitro capacitation of fresh
semen samples was performed in BO media supplemented with heparin for 2 hrs.
Genes such as SLC16A1 and SLC16A7 were selected for the study. The mRNA
isolated from respective samples were converted to cDNA and subjected to q-PCR
analysis. The target genes were normalized to endogenous control GLUT5. The
mRNA expression of genes in fresh capacitated was analyzed relative to fresh
uncapacitated as control. Fold change in gene expression was calculated using ∆∆Cq
method. Significant difference between groups for fold change was analyzed using t
test and multiple comparison was performed by Duncan multiple range test. No
significant difference (p<0.05) was observed in the mRNA expression of MCT1and
MCT2 between fresh capacitated and fresh uncapacitated (control) semen samples. It
was concluded that, in vitro capacitation induces no significant change in the mRNA
expression of both SLC16A1 and SLC16A7 genes which encode for MCT1 and MCT2
respectively and MCTs exert minimal role in capacitation related biochemical changes
in bubaline spermatozoa.