Lack of differential sperm acrosome-associated ۱ gene expression between X- and Y-bearing spermatozoa in Bali bulls (Bos javanicus domesticus)

This study aimed to determine whether the sperm acrosome-associated ۱ (SPACA۱) gene exhibits differential expression between X- and Y-bearing spermatozoa of Bali bulls and assess its potential as an RNA biomarker for sperm sexing. Sexed frozen semen (X or Y fractions) from Bali bulls was obtained from the Singosari National Artificial Insemination Center, Malang, Indonesia. Post-thaw semen quality was evaluated for motility, viability, abnormality, concentration, and plasma membrane integrity. Total RNA was extracted separately from X and Y fractions using easy-BLUE reagent, and SPACA۱ expression was quantified by RT-qPCR using GAPDH as an internal control. Relative expression was calculated by the ۲−ΔΔCt method, and group differences were analyzed with the Mann-Whitney U test. The SPACA۱ expression was numerically higher in X-bearing than Y-bearing spermatozoa (۱.۰۰ versus ۰.۵۱), but the difference was not significant. Thus, SPACA۱ transcripts do not provide a reliable molecular marker for sperm sex differentiation. Future work should focus on protein-level characterization of SPACA۱ and exploration of sex-biased surface proteins for immunological sexing. This research supports Sustainable Development Goal ۲: Zero Hunger, by contributing to reproductive efficiency in livestock, and Sustainable Development Goal ۱۲: Responsible Consumption and Production, through development of cost-effective, sustainable sperm sexing technologies.

کلیدواژه ها:

Bos javanicus domesticus ، RT-qPCR ، Sperm acrosome-associated ۱ ، sustainable reproduction ، Zero Hunger