Chronological profiling of early pregnancy transcripts in Murrah buffaloes

Effective reproductive management in cattle, such as cows and buffaloes, requires early and accurate pregnancy detection. Early identification of pregnancy enables farmers to promptly identify non-pregnant animals for treatment and/or rebreeding, thereby reducing the calving interval. This study aimed to standardize the expression of the CCL۸ and CXCL۱۰ genes as markers for early pregnancy detection in Murrah buffaloes. Blood samples were collected on the ۱۶th day post-artificial insemination for gene expression analysis and on days zero, seven, ۱۴, and ۲۱ post-artificial insemination for progesterone concentration measurement. Buffaloes were categorized as pregnant (n = ۶) or non-pregnant (n = ۶) based on the resumption of estrus. Gene expression levels in peripheral blood leukocytes were analyzed using Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) with SYBR green dye. Amplicons of CCL۸, CXCL۱۰, and GAPDH genes were measured ۱۰۸, ۱۱۷, and ۱۵۸ bp, respectively. Results showed that CCL۸ mRNA expression in pregnant buffaloes was ۵.۱۳ and ۱۲.۲۱ times higher compared to non-pregnant buffaloes, while CXCL۱۰ mRNA expression was ۴.۱۹ and ۲۲.۱۷ times higher. These findings indicated significantly elevated CCL۸ and CXCL۱۰ mRNA expression levels in peripheral blood leukocytes of pregnant buffaloes on the ۱۶th day. Progesterone levels in the pregnant group were increased significantly from day zero to day ۲۱, while no significant differences were observed between groups on days zero, seven and ۱۴. Pregnancy was further confirmed via per-rectal examination on the ۴۵th day post-artificial insemination Therefore, CCL۸ and CXCL۱۰ gene expression profiling on the ۱۶th day could serve as reliable early pregnancy markers in Murrah buffaloes.

کلیدواژه ها:

Chemokines ، Early pregnancy marker ، Gene expression ، Murrah buffaloes ، qRT-PCR