Preliminary evaluation of PRM1, TNP1, and MARCH1 transcript abundance in semen of Bali bulls

Abstract

Superior bulls with high Total Motile Spermatozoa (TMS) values can generate substantially more frozen semen doses per year, and integrating semen production evaluation with gene expression analysis may offer complementary information for evaluating breeding bulls. This study aimed to analyze semen productivity of Bali bulls and the expression of protamine 1 (PRM1), Transition Nuclear Protein 1 (TNP1), and membrane-associated ring-CH 1 (MARCH1) genes in Bali bulls. Secondary data from 28 Bali bulls at the Singosari National Artificial Insemination Center (SNAIC) were collected, including semen volume, motility, and sperm concentration, from which TMS per ejaculate was calculated. Bulls were classified into high-productivity (n=9) and low-productivity (n=3) groups based on significant TMS differences (P<0.05). RNA was extracted from frozen-thawed spermatozoa, and gene expression was quantified by Real-Time PCR using the 2−ΔΔCT method. The high-productivity group produced significantly higher average TMS (299,992±115,974 ×10⁶ sperm/year) and frozen semen straws (12,000±4,639 straws/year) than the low-productivity group (P<0.05), whereas PRM1, TNP1, and MARCH1 expression did not differ significantly between groups. These results suggest that PRM1, TNP1, and MARCH1 expression was not linked to productivity differences, likely due to limited statistical power from the small, unbalanced sample. TNP1 and MARCH1 remain candidates warranting validation in larger studies.

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