Comparative Analysis of Gonadotropin-Releasing Hormone Receptor Gene Expression in Pituitary Glands of Prolific Malabari and Non-Prolific Attappady Black Goats

A study was conducted to investigate the expression profile of gonadotropin-releasing hormone receptor (GNRHR) gene in pituitary glands of prolific Malabari and non-prolificAttappady Black goats. To ensure reliable normalization of GNRHR gene expression, stability of four candidate reference genes viz., Beta-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylprolyl isomerase A (PPIA) and ribosomal protein L34 (RPL34) were evaluated in pituitary tissues using quantitative real-time polymerase chain reaction (qPCR). Comprehensive stability analysis employing BestKeeper, delta Ct, geNorm, NormFinder and RefFinder algorithms identified GAPDH as the most stable reference gene, followed by RPL34, ACTB and PPIA. Normalisation to GAPDH revealed that relative GNRHR expression was significantly higher (1.64 fold, P=0.018) in Malabari goats than Attappady Black goats. Furthermore, in Malabari goats, heterozygous diplotypes had significantly higher GNRHR expression (1.49 fold, P=0.02) than homozygous diplotypes. This study establishes GAPDH as a robust reference gene for accurate GNRHR gene expression analysis in goat pituitary tissues, providing a foundational framework for future investigations into reproductive gene regulation in small ruminants.