Veterinary Research Forum

LACK OF DIFFERENTIAL SPACA1 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
Viski Fitri Hendrawan 2
Suryo Kuncorojakti 3
Suherni Susilowati 3
Muhammad Yusuf 4
Sahiruddin Sahiruddin 4
Masturi Masturi 4
Athhar Manabi Diansyah 4
Sigit Bintara 5
Aeni Nurlatifah 5
Yayuk Kholifah 6
Riski Lesta Mega 3
Devia Yoanita Kurniawati 3
Intan Firdha Olien Nor Al Ichsan 3

1 Department of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya

2 Faculty of Veterinary Medicine, Brawijaya University, Jl. Veteran No.10–11, Malang 65145, Indonesia

3 Faculty of Veterinary Medicine, Universitas Airlangga, Campus C, Jl. Mulyorejo, Surabaya 60115, Indonesia

4 Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan Km.10, Tamalanrea, Makassar 90245, Indonesia

5 Faculty of Animal Science, Universitas Gadjah Mada, Jl. Fauna No. 3, Bulaksumur, Yogyakarta 55281, Indonesia

6 Balai Besar Inseminasi Buatan Singosari (Singosari National Artificial Insemination Center), Jl. BBIB No.1, Desa Toyomarto, Kec. Singosari, Malang 65153, 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 to 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. 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 vs. 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 SDG 2 (Zero Hunger) by contributing to reproductive efficiency in livestock, and SDG 12 (Responsible Consumption and Production) through development of cost-effective, sustainable sperm sexing technologies.

Keywords

Bali cattle
SPACA1
spermatozoa
RT-qPCR
sustainable reproduction
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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