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Effects of insulin-like growth factor-1 on the proliferation and apoptosis of stallion testicular cells under normal and heat stress culture conditions.
Abstract: This study investigated the effect of heat stress on stallion testicular cells (TCs) and the effect of insulin-like growth factor (IGF)-1 on TC viability, proliferation, and apoptosis, including different stages of germ cells. TCs were divided into control or treatment groups with 0.01, 0.1, 1, 10, and 100 ng/mL of recombinant human IGF-1 (rhIGF-1) for 24 h at 34 °C and 37 °C. The population and viability were measured before and after treatment. The effects of rhIGF-1 on TC viability, proliferation, and apoptosis were determined using RT-qPCR. Proliferating cell nuclear antigen (PCNA) and marker of proliferation Ki-67 (MKI-67) were used as proliferation markers. Myeloid leukemia-1 (MCL-1) was used as an antiapoptotic marker. BCL2 antagonist/killer-1 (BAK-1) was used as a proapoptotic marker. The relative abundance of mRNA transcript of undifferentiated cell transcription factor 1 (UTF-1), protein gene product 9.5 (PGP9.5), and deleted in azoospermia-like (DAZL), was measured for spermatogenesis progression. TCs treated with 1 ng/mL rhIGF-1 at 34 °C exhibited the highest viability. Significant upregulation of the relative abundance of mRNA transcript of PCNA, MKI-67, and MCL-1 was observed in treated TCs compared with untreated TCs; however, BAK-1 was significantly downregulated in treated TCs. Germ cells treated with 1 ng/mL rhIGF-1 exhibited the highest relative abundance of mRNA transcript of UTF-1 and DAZL, whereas TCs exposed to 0.1 ng/mL showed the highest PGP9.5 level. These data confirm that heat stress in stallions decreases TC viability. These findings may help identify a basal IGF-1 level for TC proliferation and apoptosis during heat stress-induced testicular degeneration in stallions.
Copyright © 2023. Published by Elsevier B.V.
Publication Date: 2023-08-15 PubMed ID: 37633109DOI: 10.1016/j.anireprosci.2023.107319Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research investigates the impact of heat stress on the health of stallion testicular cells, and how insulin-like growth factor-1 can potentially enhance their viability and proliferation, and hinder apoptosis. This study can aid in understanding the base level of IGF-1 required for maintaining health of testicular cells during heat stress-induced degeneration.
Methods and Approach
- The investigators chose to use stallion testicular cells (TCs) for their research, assessing the effect of heat and insulin-like growth factor (IGF)-1 on these cells.
- The TCs were divided into control and treatment groups that were exposed to differing concentrations of recombinant human IGF-1 for 24 hours at either 34 degrees Celsius or 37 degrees Celsius.
- Quantitative PCR (RT-qPCR) was applied to measure the effects of IGF-1 on TC proliferation, viability, and apoptosis.
- The researchers used two proliferation markers, proliferating cell nuclear antigen (PCNA) and marker of proliferation Ki-67 (MKI-67), and the anti-apoptotic marker, Myeloid leukemia-1 (MCL-1), to gauge cell proliferation and survival. A pro-apoptotic marker, BCL2 antagonist/killer-1 (BAK-1), was used to measure cell death.
- They also used some gene tests to study progression of spermatogenesis: undifferentiated cell transcription factor 1 (UTF-1), protein gene product 9.5 (PGP9.5), and deleted in azoospermia-like (DAZL).
Results and Findings
- The study concluded that TCs treated with 1 ng/mL of IGF-1 at 34 degrees Celsius had the highest survival rate.
- A significant increase in the relative abundance of PCNA, MKI-67, and MCL-1 mRNA transcript was registered in the treated TCs when compared to the control group. On the other hand, BAK-1 was significantly downregulated in the treated group, indicating lower rate of apoptosis.
- TCs that were treated with 1 ng/mL IGF-1 showed the most substantial relative abundance of UTF-1 and DAZL. This indicates that IGF-1 treatment could aid in promoting spermatogenesis.
- Testicular cells exposed to 0.1 ng/mL IGF-1 exhibited the most pronounced PGP9.5 level, signifying the presence of spermatogonia and primary spermatocytes within these cells.
- The results confirmed that heat stress in stallions decreases the survival of their testicular cells.
Implications
- The data gathered from this research can be helpful in identifying the necessary base level of IGF-1 that would enhance proliferation and reduce apoptosis in TCs during heat stress-induced testicular degeneration in stallions.
- These findings may pave the way for treatments or interventions that can mitigate the adverse effects of heat stress on stallion fertility.
Cite This Article
APA
Shakeel M, Yoon M.
(2023).
Effects of insulin-like growth factor-1 on the proliferation and apoptosis of stallion testicular cells under normal and heat stress culture conditions.
Anim Reprod Sci, 256, 107319.
https://doi.org/10.1016/j.anireprosci.2023.107319 Publication
Researcher Affiliations
- Department of Animal Science and Biotechnology, Kyungpook National University, Sangju 37224, Republic of Korea; Department of Clinical Studies, Faculty of Veterinary and Animal Sciences, Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi 44000, Pakistan.
- Department of Animal Science and Biotechnology, Kyungpook National University, Sangju 37224, Republic of Korea; Department of Horse, Companion, and Wild Animal Science, Kyungpook National University, Sangju 37224, Republic of Korea; Research Centre for Horse Industry, Kyungpook National University, Sangju 37224, Republic of Korea. Electronic address: mjyoon@knu.ac.kr.
Conflict of Interest Statement
Declaration of Competing Interest We wish to confirm that there are no known conflicts of interest associated with this publication.
Citations
This article has been cited 4 times.- Ghaleghafi A, Khorshidi KJ, Jafari MA. Nutritional and managerial interventions on growth performance, nutrient transport-related genes and behavioral responses in heats stressed broiler chickens. Vet Anim Sci 2026 Mar;31:100561.
- Shakeel M, Yoon M. Brain-derived neurotrophic factor and neurotrophic tyrosine receptor kinase-2 in stallion testes: insights into seasonal changes and potential roles in spermatogenesis. J Anim Sci Technol 2025 Jul;67(4):909-921.
- Fukui T, Terashima A, Omata Y, Chijimatsu R, Okamoto K, Tsukasaki M, Fukuda Y, Hayata T, Saitoh A, Toda E, Takayanagi H, Tanaka S, Terashima Y, Saito T. Disulfiram ameliorates bone loss in ovariectomized mice by suppressing osteoclastogenesis. J Bone Miner Metab 2025 Mar;43(2):61-73.
- Khan MZ, Khan A, Chen W, Chai W, Wang C. Advancements in Genetic Biomarkers and Exogenous Antioxidant Supplementation for Safeguarding Mammalian Cells against Heat-Induced Oxidative Stress and Apoptosis. Antioxidants (Basel) 2024 Feb 20;13(3).
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