Pituitary extract supplementation enhances follicular survival and gonadotropin receptor expression in vitrified ovarian tissue of Aceh cattle, Indonesia

Vet World Vol.18 November-2025 Article - 23

Research Article

Veterinary World, 18(11): 3607-3621

https://doi.org/10.14202/vetworld.2025.3607-3621

Cut Intan Novita^1,2, Tongku Nizwan Siregar³, Ni Wayan Kurniani Karja⁴, Sri Wahyuni⁵, and Amalia Sutriana⁶

1. Animal Sciences Study Program, Faculty of Agriculture, Universitas Syiah Kuala, Banda Aceh, Indonesia.

2. Graduate School of Mathematics and Applied Sciences, Universitas Syiah Kuala, Banda Aceh, Indonesia.

3. Laboratory of Reproduction, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia.

4. Division of Reproduction and Obstetrics, School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia.

5. Laboratory of Anatomy, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia.

6. Laboratory of Pharmacology, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia.

Background and Aim: Vitrification is a promising cryopreservation technique for conserving the genetic resources of Aceh cattle; however, it may induce cryoinjury and follicular apoptosis. Pituitary extract (PE), containing follicle-stimulating hormone (FSH) and luteinizing hormone (LH), can potentially mitigate apoptosis and maintain follicular viability. This study investigated the effect of bovine PE supplementation in vitrification media on the expression of caspase-3, FSH receptor (FSHR), and LH receptor (LHR) in vitrified ovarian tissue of Aceh cattle.

Materials and Methods: A factorial, completely randomized design was employed using five vitrification media, PE at 0, 200, 400, and 600 μg/mL, and commercial bovine PE (Gibco, 300 μg/mL), combined with three vitrification durations (0, 7, and 14 days), each in triplicate. Post-vitrification ovarian tissue was evaluated by immunohistochemistry for caspase-3, FSHR, and LHR expression. The intensity scores (IS) were analyzed using the Kruskal–Wallis and Mann–Whitney U tests (p < 0.05).

Results: PE supplementation significantly (p < 0.05) reduced caspase-3 expression across all follicular stages, indicating decreased apoptosis, whereas vitrification duration alone showed limited effects. The lowest caspase-3 IS (negative) occurred in the 600 μg/mL PE group after 7 days. Both PE concentration and vitrification duration significantly influenced FSHR and LHR expression (p < 0.05), with strong positive interactions between the two factors. FSHR and LHR expression increased progressively from primordial to antral follicles, suggesting preserved gonadotropin responsiveness. Optimal follicular preservation and receptor integrity were achieved at 600 μg/mL PE following 7 days of vitrification.

Conclusion: Supplementation of vitrification media with 600 μg/mL bovine PE effectively suppresses follicular apoptosis while enhancing FSHR and LHR expression, maintaining ovarian functionality after vitrification. These results highlight PE as a low-cost, multi-hormonal alternative to single-gonadotropin supplements in cryopreservation media, supporting sustainable genetic conservation of Aceh cattle. Integrating locally derived PE into cryopreservation protocols could improve post-thaw follicular survival, reduce dependency on commercial reagents, and strengthen reproductive biotechnology for indigenous livestock conservation.

Keywords: Aceh cattle, caspase-3, cryopreservation, follicle-stimulating hormone receptor, luteinizing hormone receptor, pituitary extract, vitrification.

How to cite this article: Novita CI, Siregar TN, Karja NWK, Wahyuni S, and Sutriana A (2025) Pituitary extract supplementation enhances follicular survival and gonadotropin receptor expression in vitrified ovarian tissue of Aceh cattle, Indonesia, Veterinary World, 18(11): 3607–3621.

Received: 11-07-2025 Accepted: 28-10-2025 Published online: 29-11-2025

Corresponding author: Tongku Nizwan Siregar E-mail: siregar@usk.ac.id

DOI: 10.14202/vetworld.2025.3607-3621

Copyright: Novita, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.