Veterinary Research Forum

Lack of differential sperm acrosome-associated 1 gene expression between X- and Y-bearing spermatozoa in Bali bulls (Bos javanicus domesticus)

Articles in Press

Document Type : Original Article

Authors

Widjiati Widjiati 1
Suryo Kuncorojakti 1
Suherni Susilowati 1
Muhammad Yusuf 2
Sahiruddin Sahiruddin 2
Masturi Masturi 2
Athhar Manabi Diansyah 2
Sigit Bintara 3
Aeni Nurlatifah 3
Yayuk Kholifah 4
Riski Lesta Mega 1
Devia Yoanita Kurniawati 1
Viski Fitri Hendrawan 5
Intan Firdha Olien Noor Al Ichsan 1

1 Department of Veterinary Reproduction, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia

2 Department of Animal Production, Faculty of Animal Science, Hasanuddin University, Makassar, Indonesia

3 Department of Animal Breeding and Reproduction, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta, Indonesia

4 Balai Besar Inseminasi Buatan Singosari (BBIB Singosari), Directorate General of Livestock and Animal Health Services, Ministry of Agriculture of the Republic of Indonesia, Malang, Indonesia

5 Department of Veterinary Reproduction, Faculty of Veterinary Medicine, Universitas Brawijaya, Malang, Indonesia

10.30466/vrf.2026.2077333.4986
Abstract
This study aimed to determine whether the sperm acrosome-associated 1 (SPACA1) 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 SPACA1 expression was quantified by RT-qPCR using GAPDH as an internal control. Relative expression was calculated by the 2−ΔΔCt method, and group differences were analyzed with the Mann-Whitney U test. The SPACA1 expression was numerically higher in X-bearing than Y-bearing spermatozoa (1.00 versus 0.51), but the difference was not significant. Thus, SPACA1 transcripts do not provide a reliable molecular marker for sperm sex differentiation. Future work should focus on protein-level characterization of SPACA1 and exploration of sex-biased surface proteins for immunological sexing. This research supports Sustainable Development Goal 2: Zero Hunger, by contributing to reproductive efficiency in livestock, and Sustainable Development Goal 12: Responsible Consumption and Production, through development of cost-effective, sustainable sperm sexing technologies.

Keywords

Bos javanicus domesticus
RT-qPCR
Sperm acrosome-associated 1
Sustainable reproduction
Zero Hunger
Subjects
Veterinary Research Forum

Articles in Press, Accepted Manuscript
Available Online from 20 July 2026

  • Receive Date 18 November 2025
  • Revise Date 17 February 2026
  • Accept Date 25 April 2026

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