EXPRESSION PROFILE OF OASl AND MX2 GENES IN PERIPHERAL BLOOD AND LUTEAL CELLS FOR EARLY PREGNANCY DIAGNOSIS IN CATTLE
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Date
2021
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Abstract
The present study was planned to analyse the expression levels of OAS 1 and
MX2 genes in Peripheral Blood Mononuclear Cells (PBMC) and luteal cell culture
upon stimulation of Interferon tau (IFNT). 30 cows were selected and divided into 2
groups as group 1 (n^lO) and II (n=20). Group I cows served as control and group 11
cows were synchronized with CIDR + PGF2a protocol in the presence of matured CL.
Breeding was carried during the observed estrus in group 1 and 48 and 72 h after
removal of CIDR in group II cows. The blood samples were collected on day 0, 7,
14, 18 and 21 post insemination and PBMC harvested was utilized for isolation and
synthesis of RNA and cDNA. Quantitative real time PCR was carried out for OASl
and MX2 genes and relative expression was calculated by 2’^^ method. Buffalo
ovaries with corpus luteum (CL) were collected immediately following slaughter.
Mid luteal phase CL was selected and cell culture was carried out and incubated for
72 h. Cultured luteal cells were treated with three early pregnant serum which was
collected on 24^ day post Al from group II cows and stored at -80° C. Varying
concentrations of Interferon tau (IFNT) as 1, 10 and 100 ng/ml were also exposed to
cultured luteal cells and incubated for 48 h and cells were harvested for RNA
isolation followed by cDNA synthesis. Quantitative real time PCR was carried out
for OASl and MX2 genes and relative expression was calculated by 2 2 -^^CT method.
The results concluded that cows treated with CIDR + PGFaa showed 100 per
cent estrus induction response and achieved 60.00 per cent of conception rate.
Expression of both OASl and MX2 genes in PBMC was significantly higher
(P<0.01) on day 14, 18 and 21 in group I and 11 pregnant cows, when compared to
group I and 11 non pregnant cows. No significant difference was observed in the
relative expression of OASl between different days in non pregnant cows of group I
and II and between different days in pregnant cows of group 1 and IL In luteal cell
culture, the OASl and MX2 expression was significantly higher (P<0.05) in early
pregnant serum and IFNT treated luteal cells when compared to untreated control
luteal cells. The OASl and MX2 expression was significantly higher (P<0.()5) in 10
and 100 ng/ml IFNT treated luteal cells when compared to Ing/ml IFNT treated
luteal cells. Hence, it could be inferred that the higher expression of OAS1 and MX2
genes during early pregnancy corresponds to the translation of related proteins, which
if identified could be used as a bio marker for early pregnancy diagnosis in cattle.