HomeEquine Research BankAndrologyEffect of testicular degeneration on expression of sperm pro...

Andrology2023; doi: 10.1111/andr.13464

Effect of testicular degeneration on expression of sperm protein at 22 kDa in stallions.

Miller, LMJ
·
Woodward, EM
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Campos, JR
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Squires, EL
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Troedsson, MHT

Abstract: Sperm protein at 22 kDa has been associated with fertility. Objective: The objectives of this study were to determine (1) the localization pattern of SP22 on ejaculated and caudal epididymal equine spermatozoa and in epididymal fluid, and to (2) characterize SP22 protein and mRNA expression in testicular and epididymal tissues in response to heat-induced testicular degeneration. Methods: Semen was collected before and after hemi-castration, as well as prior to and following insulation of the remaining testes, and tissue specimens were collected for analysis. Results: Histopathology confirmed degeneration in insulated testes. Ejaculated and epididymal spermatozoa from samples collected prior to insulation of the testicles had a predominant staining pattern of SP22 over the equatorial region. However, the equatorial pattern in the pre-insulation epididymal semen samples was significantly lower than in the pre-insulation ejaculated semen samples (68 ± 3, 81 ± 2.6, respectively). Ejaculated and epididymal samples collected after insulation of the testicles showed a complete loss of staining as the predominant pattern. Western blot analysis verified the presence of SP22 on fresh ejaculated spermatozoa prior to and following heat-induced degeneration, on epididymal spermatozoa after testicular insulation, and in testicular and epididymal tissues. Heat insulation significantly reduced messenger RNA expression in the head of the epididymis and testicular tissues. Immunohistochemistry of the testicular and epididymal tissues pre-heating showed considerably weaker staining than the same tissues post-heating. Conclusions: It was concluded that heat-induced testicular damage causes both loss and relocation of SP22 on the sperm membrane. Future studies are warranted to determine the diagnostic value of these findings.

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Publication Date: 2023-05-23 PubMed ID: 37220913DOI: 10.1111/andr.13464Google Scholar: Lookup

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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study explores how heat-induced testicular degeneration in stallions affects the expression and location of the sperm protein SP22, which is associated with fertility. The research involved collection and analysis of semen before and after testicular insulation and hemi-castration, with findings indicating that heat-induced damage causes both loss and relocation of SP22 on the sperm membrane.

Study Objectives and Methodology

The study had two main objectives: to identify where the sperm protein SP22 is located on both ejaculated and epididymal equine sperm and in epididymal fluid, and to characterize how SP22 protein and mRNA expression in the testes and epididymal tissues changes due to heat-induced testicular degeneration.
Methods involved collecting semen before and after hemi-castration, and also before and after testicular insulation. Tissue samples were also collected for analysis.

Results and Findings

Testicular degeneration in insulated testes was confirmed through histopathology.
Prior to insulation, SP22 was predominantly found in the equatorial region of ejaculated and epididymal sperm. However, there was a significant decrease in this pattern in the pre-insulation epididymal semen compared to the ejaculated semen.
After testicular insulation, both ejaculated and epididymal samples showed a complete loss of SP22 staining.
Western blotting confirmed the presence of SP22 on fresh ejaculated sperm both before and after heat-induced degeneration, on epididymal sperm after testicular insulation, and in testicular and epididymal tissues.
The heat insulation led to a significant decrease in mRNA expression in the head of the epididymis and in the testes.
Immunohistochemical analysis showed considerably weaker staining in testicular and epididymal tissues before heating than after.

Conclusions

The study concluded that heat-induced testicular damage results in both a loss and relocation of SP22 on the sperm membrane.
The findings from this study suggest that further research is required to determine the diagnostic value of these observations.

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APA

Miller LMJ, Woodward EM, Campos JR, Squires EL, Troedsson MHT. (2023). Effect of testicular degeneration on expression of sperm protein at 22 kDa in stallions. Andrology. https://doi.org/10.1111/andr.13464

Publication

Andrology

ISSN: 2047-2927

NlmUniqueID: 101585129

Country: England

Language: English

Researcher Affiliations

Miller, Lynda M J

Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky, USA.
College of Veterinary Medicine, Lincoln Memorial University, Harrogate, Tennessee, USA.

Woodward, Elizabeth M

Department of Clinical Studies, New Bolton Center, University of Pennsylvania School of Veterinary Medicine, Kennett Square, Pennsylvania, USA.

Campos, Juliana R

Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky, USA.

Squires, Edward L

Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky, USA.

Troedsson, Mats H T

Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky, USA.

References

This article includes 34 references

Nagabuko D, Taira T, Kitaura H, etu00a0al. DJ-1, a novel oncogene which transforms mouse NIH3T3 cells in cooperation with ras. Biochem Biophys Res Commun. 1997;23:509-513.
Bonifati V, Rizzu P, Van Baren MJ, etu00a0al. Mutations in the DJ-1 gene associated with autosomal recessive early onset Parkinsonism. Science. 2003;299:256-259.
Klinefelter GR, Laskey JW, Ferrell J, Suarez JD, Roberts NL. Discriminant analysis indicates a single sperm protein (SP22) is predictive of fertility following exposure to epididymal toxicants. J Androl. 1997;18:139-150.
De Lima Rosa J, Figueiredo TM, Barros JWF, etu00a0al. SP22 sperm protein as a potential biomarker of fertility in humans: a preliminary study. Reprod Toxicol. 2023;117:108343.
Klinefelter GR, Welch JE, Perreault SD, etu00a0al. Localization of the sperm protein SP22 and inhibition of fertility in vivo and in vitro. J Androl. 2002;23:48-63.
Okada M, Matsumoto K, Niki T, Taira T, Iguchi-Ariga SMM, Ariga H. DJ-1, a target protein for an endocrine disrupter, participates in the fertilization in mize. Biol Pharm Bull. 2002;25:853-856.
Miller LMJ, Troedsson MHT, Duoos LA, Klinefelter GR, Roberts KP. Immunocytochemical detection and localization of sperm protein 22 (SP22) in fresh and cryopreserved equine semen. Biol Reprod. 2003;68:166-167.
Miller LMJ, Wells BA, Macpherson ML, Roberts KP, Troedsson MHT. Effect of cryopreservation and extenders on the expression and localization of sperm protein at 22u00a0kDa (SP22) on equine spermatozoa. Theriogenology. 2005;64:779-779.
Miller L, Woodward E, Campos J, Squires E, Troedsson M. Distribution pattern(s) of sperm protein at 22u00a0kDa (SP22) on fresh, cooled and frozen/thawed equine spermatozoa and expression of SP22 in tissues from the testes and epididymides of normal stallions. Reprod Domest Anim. 2015;50:275-282.
Miller LMJ, Greene ES, Roberts KP, Troedsson MHT. Expression of equine sperm protein 22u00a0kDa (SP22) in testicular and epididymal tissue. Anim Reprod Sci. 2006;94:54-55.
Blanchard TL, Varner DD. Testicular degeneration. In: Mckinnon, AO, Voss, JL, eds. Equine Reproduction. Lea & Febiger; 1993.
Freidman R, Scott M, Heath SE, Hughes JP, Daels PF, Tran TQ. The effects of increase testicular temperature on spermatogenesis in the stallion. J Reprod Fertil Suppl. 1991;44:127-134.
Harrison R, Weiner J. Abdomino-testicular temperature gradients. J Physiol. 1948;18:256-262.
Ulberg LC. The influence of high temperature on reproduction. J Hered. 1958;49:62-64.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402-408.
Glover TD. Some effects of scrotal insulation on the semen of rams. Proc Soc Study Fertil. 1955;7:66-75.
Glover TD. The effect of scrotal insulation and the influence of the breeding season upon fructose concentration in the semen of the ram. J Endocrinol. 1956;13:235-242.
Byers SW, Glover TD. Effect of scrotal insulation on the pituitary-testicular axis of the ram. J Reprod Fertil. 1984;71:23-31.
Mieusset R, Sowerbutts SF, Zupp JL, Setchell BP. Increased flow of testicular blood plasma during local heating of the testes of rams. J Reprod Fertil. 1992;94:345-352.
Ross AD, Entwistle KW. The effect of scrotal insulation on spermatozoal morphology and the rates of spermatogenesis and epididymal passage of spermatozoa in the bull. Theriogenology. 1979;11:111-129.
Prabhakar J, Chimbombi M, Malmgren L, Fredriksson G, Madej A. Effects of testosterone and LH concentrations of induced testicular degeneration in bulls. Acta Vet Scand. 1990;31:505-507.
Sidibe M, Franco LA, Fredriksson G, Madej A, Malmgren L. Effects on testosterone, LH and cortisol concentrations, and on testicular ultrasonographic appearance of induced testicular degeneration in bulls. Acta Vet Scand. 1992;33:191-196.
Karabinus DS, Vogler CJ, Saacke RG, Evenson DP. Chromatin structural changes in sperm after scrotal insulation of Holstein bulls. J Androl. 1997;18:549-555.
Malmgren L, Larsson K. Experimentally induced testicular alterations in boars: histological and ultrastructural findings. Zentralbl Veterinarmed A. 1989;36:3-14.
Malmgren L. Experimentally induced testicular alterations in boars: hormonal changes in mature and peripubertal boars. Acta Vet Scand. 1990;31:97-107.
Robinson BR, Netherton JK, Ogle RA, Baker MA. Testicular heat stress, a historical perspective and two postulates for why germ cells are heat sensitive. Biol Rec. 2023;98:603-622.
Welch JE, Barbee RR, Roberts NL, Suarez JD, Klinefelter GR. SP22: a novel fertility protein from a highly conserved gene family. J Androl. 1998;19:385-393.
Pena FJ. Detecting subtle changes in sperm membranes in veterinary andrology. Asian J Androl. 2007;9:731-737.
Lee LP, Fritz IB. Studies on spermatogenesis in rats. V. Increased thermal lability of lysosomes from testicular germinal cells and its possible relationship to impairments in spermatogenesis in cryptorchidism. J Biol Chem. 1972;247:7956-7961.
Lee LP. Temperature effect on the permeability of plasma membranes of advanced germinal cells of the rat testis. Can J Biochem. 1974;52:586-593.
Rochwerger L, Cohen DJ, Cuasnicu PS. Mammalian sperm-egg fusion: the rat egg has complementary sites for a sperm protein that mediates gamete fusion. Dev Biol. 1992;153:83-90.
Morrissette J, Kratzschmar J, Haendler B, etu00a0al. Primary structure and properties of helothermine, a peptide toxin that blocks ryanodine receptors. Biophys J. 1995;68:2280-2288.
Milne TJ, Abbenante G, Tyndall JD, Halliday J, Lewis RJ. Isolation and characterization of a cone snail protease with homology to CRISP proteins of the pathogenesis-related protein superfamily. J Biol Chem. 2003;278:31105-31110.
Klinefelter GR, Roberts NL, Suarez JD. Direct effects of ethane dimethanesulphonate on epididymal function in adult rats. An in vitro demonstration. J Androl. 1992;13:409-421.

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