Insulin-like growth factor-I (IGF-I) protects cultured equine Leydig cells from undergoing apoptosis.
Abstract: Leydig cells located in the interstitial space of the testicular parenchyma produce testosterone which plays a critical role in the maintenance and restoration of spermatogenesis in many species, including horses. For normal spermatogenesis, maintaining Leydig cells is critical to provide an optimal and constant level of testosterone. Recently, an anti-apoptotic effect of IGF-I in testicular cells in rats has been reported, but a similar effect of IGF-I on equine Leydig cells remains to be elucidated. If IGF-I also protects stallion testicular cells from undergoing apoptosis, then IGF-I may have potential as a treatment regime to prevent testicular degeneration. The present study was designed to evaluate the anti-apoptotic effect of IGF-I on cultured equine Leydig cells. Testes were collected from 5 post-pubertal stallions (2-4 years old) during routine castrations. A highly purified preparation of equine Leydig cells was obtained from a discontinuous Percoll gradient. Purity of equine Leydig cells was assessed using histochemical 3β-HSD staining. Equine Leydig cells and selected doses of recombinant human IGF-1 (rhIGF-I; Parlow A.F., National Hormone and Peptide Program, Harbor-UCLA Medical Center) were added to wells of 24 or 96 well culture plates in triplicate and cultured for 24 or 48 h under 95% air:5% CO(2) at 34°C. After 24 or 48 h incubation, apoptotic rate was assessed using a Cell Death Detection ELISA kit. Significantly lower apoptotic rates were observed in equine Leydig cells cultured with 5, 10, or 50ng/ml of rhIGF-I compared with control cells cultured without rhIGF-I for 24h. Exposure to 1, 5, 10 or 50 ng/ml of rhIGF-I significantly decreased apoptotic rate in equine Leydig cells cultured for 48 h. After 48 h incubation, cells were labeled with Annexin V and propodium iodine to determine the populations of healthy, apoptotic, and necrotic cells by counting stained cells using a Nikon Eclipse inverted fluorescence microscope. As a percentage of the total cells counted, significantly lower numbers of apoptotic cells were observed in cells treated with 10 (9%) or 50 ng/ml (10%) of rhIGF-I compared with cells cultured without rhIGF-I (control, 22%). In this study, the results from the two assays indicated that rhIGF-I protected equine Leydig cells from undergoing apoptosis during cell culture for 24h or 48 h. In conclusion, IGF-I may be an important paracrine/autocrine factor in protecting equine Leydig cells from undergoing apoptosis.
Copyright © 2010 Elsevier B.V. All rights reserved.
Publication Date: 2010-10-21 PubMed ID: 21071158DOI: 10.1016/j.anireprosci.2010.10.001Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research explores the potential protective effect of Insulin-like growth factor-I (IGF-I) on horse Leydig cells, which are integral to the production of testosterone and sperm generation, to prevent them from naturally dying or undergoing apoptosis.
Introduction
The study focuses on the effect of IGF-I on Leydig cells. These cells, found in horses’ testes, are responsible for producing testosterone – a vital hormone for spermatogenesis or the production of sperm. The researchers have used previous studies that demonstrated IGF-I’s anti-apoptotic effect on rat testicular cells as a base, and sought to determine if a similar effect could be found in horses.
Materials and Methods
- The study used testes from 5 post-pubertal stallions, aged 2-4 years, as the cell source.
- Leydig cells were extracted using a discontinuous Percoll gradient, and their purity was verified via 3β-HSD staining.
- The purified Leydig cells were cultured in plates, in both 24 and 48 hour periods, using different doses of recombinant human IGF-I (rhIGF-I).
- The effect of rhIGF-I on apoptosis was measured using a Cell Death Detection ELISA kit.
Results
- The researchers observed significantly lower rates of apoptosis in the Leydig cells cultured with rhIGF-I compared to those cultured without rhIGF-I at both 24 and 48 hours.
- Further examination using Annexin V and propodium iodine to stain the cells revealed that fewer cells were going through apoptosis when treated with 10 or 50 ng/ml doses of rhIGF-I as compared to untreated control cells.
Conclusion
The findings confirm that rhIGF-I can protect equine Leydig cells from apoptosis. The implications of this are vast and suggest IGF-I could function as a critical factor in protecting Leydig cells in horses, potentially preventing testicular degeneration, and maintaining healthy testosterone and sperm production levels. Further research is necessary to establish how IGF-I could be utilized for this purpose effectively.
Cite This Article
APA
Yoon MJ, Roser JF.
(2010).
Insulin-like growth factor-I (IGF-I) protects cultured equine Leydig cells from undergoing apoptosis.
Anim Reprod Sci, 122(3-4), 353-358.
https://doi.org/10.1016/j.anireprosci.2010.10.001 Publication
Researcher Affiliations
- Department of Animal Science, University of California, 1 Shields Ave, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Annexin A5
- Apoptosis / drug effects
- Cell Count
- Cell Separation / veterinary
- Cells, Cultured
- Coloring Agents
- DNA Fragmentation
- Horses
- Insulin-Like Growth Factor I / pharmacology
- Leydig Cells / cytology
- Leydig Cells / drug effects
- Male
- Propidium
- Recombinant Proteins / pharmacology
- Staining and Labeling / veterinary
Citations
This article has been cited 12 times.- Salvaleda-Mateu M, Rodríguez-Varela C, Labarta E. Do Popular Diets Impact Fertility?. Nutrients 2024 May 31;16(11).
- Liu M, Liu Y, Pei LG, Zhang Q, Xiao H, Chen YW, Wang H. Prenatal dexamethasone exposure programs the decreased testosterone synthesis in offspring rats by low level of endogenous glucocorticoids. Acta Pharmacol Sin 2022 Jun;43(6):1461-1472.
- Catalano-Iniesta L, Sánchez-Robledo V, Iglesias-Osma MC, García-Barrado MJ, Carretero-Hernández M, Blanco EJ, Vicente-García T, Burks DJ, Carretero J. Sequential testicular atrophy involves changes in cellular proliferation and apoptosis associated with variations in aromatase P450 expression levels in Irs-2-deficient mice. J Anat 2019 Feb;234(2):227-243.
- Tangyuenyong S, Sato F, Nambo Y, Murase H, Endo Y, Tanaka T, Nagaoka K, Watanabe G. Comparison of physical body growth and metabolic and reproductive endocrine functions between north and south climates of Japan in trained Thoroughbred yearling horses. J Equine Sci 2017;28(3):77-86.
- Lee WY, Do JT, Park C, Kim JH, Chung HJ, Kim KW, Gil CH, Kim NH, Song H. Identification of Putative Biomarkers for the Early Stage of Porcine Spermatogonial Stem Cells Using Next-Generation Sequencing. PLoS One 2016;11(1):e0147298.
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- Piotrowska K, Sluczanowska-Glabowska S, Kucia M, Bartke A, Laszczynska M, Ratajczak MZ. Histological changes of testes in growth hormone transgenic mice with high plasma level of GH and insulin-like growth factor-1. Folia Histochem Cytobiol 2015;53(3):249-58.
- Yoon M, Jiang J, Chung KH, Roser JF. Immunolocalization of insulin-like growth factor-I (IGF-I) and its receptors (IGF-IR) in the equine epididymis. J Reprod Dev 2015;61(1):30-4.
- Jung H, Roser JF, Yoon M. UTF1, a putative marker for spermatogonial stem cells in stallions. PLoS One 2014;9(10):e108825.
- Ueno H, Hattori T, Kumagai Y, Suzuki N, Ueno S, Takagi H. Alterations in the corneal nerve and stem/progenitor cells in diabetes: preventive effects of insulin-like growth factor-1 treatment. Int J Endocrinol 2014;2014:312401.
- Afeiche MC, Bridges ND, Williams PL, Gaskins AJ, Tanrikut C, Petrozza JC, Hauser R, Chavarro JE. Dairy intake and semen quality among men attending a fertility clinic. Fertil Steril 2014 May;101(5):1280-7.
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