VASA (DDX4) is a Putative Marker for Spermatogonia, Spermatocytes and Round Spermatids in Stallions.
Abstract: Expression of the protein DDX4/MVH, or VASA, has been reported in germ cells of several species. The main objectives of this study were to (i) investigate VASA expression patterns in testicular cells of stallions at two different reproductive stages (pre-pubertal and post-pubertal) and (ii) evaluate the use of VASA antibody as a molecular marker for single germ cells from stallions. Testicular tissues were obtained from stallions and categorized as pre-pubertal and post-pubertal based on the formation of lumen and status of spermatogenesis on the cross section of seminiferous tubules. The results of Western blot showed a VASA protein band located at 76 kDa, indicating that the rabbit antibody has a cross-reactivity with horse testicular tissues. The result of immunolabelling showed that VASA was expressed in the cytoplasm of spermatogonia at both reproductive stages and in spermatocytes and round spermatids at the post-pubertal stage. GATA4-positive Sertoli cells and Leydig cells located in the interstitial space were not immunolabelled with VASA. These results suggest that VASA can be utilized as a molecular marker for germ cells of stallions at pre-pubertal and post-pubertal stages. Interestingly, immunolabelling intensity was significantly higher in pachytene spermatocytes compared to spermatogonia and round spermatid. VASA antibody was also effective for staining of single germ cell preparations. In conclusion, VASA protein expression can be used as a marker for identification of spermatogonia, spermatocytes and round spermatids in testicular tissues of stallions.
© 2015 Blackwell Verlag GmbH.
Publication Date: 2015-10-20 PubMed ID: 26482643DOI: 10.1111/rda.12632Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research examines the pattern of VASA protein expression in the testicular cells of pre-pubertal and post-pubertal stallions. The researchers suggest the potential of VASA, or DDX4/MVH, as a molecular marker for identifying different stages of germ cells in horse testicular tissues.
Understanding the Experiment
- The researchers primarily aimed to observe how VASA protein expression patterns changed in the testicular cells of stallion in different reproductive stages – the pre-pubertal and post-pubertal stages. They also evaluated the effectiveness of a VASA antibody as a molecular marker for individual germ cells within the stallions.
- The experimental design involved obtaining testicular tissues from stallions and categorizing them as either pre-pubertal or post-pubertal, based on the formation of lumen and the status of spermatogenesis as seen in the cross section of the seminiferous tubules.
- A Western blot test, a widely used analytical technique used to detect specific proteins in a sample, revealed a VASA protein band located at 76 kDa. This placed evidence that the used rabbit antibody cross-reacts with horse testicular tissues.
Results and Findings
- The study found that VASA protein was expressed in the cytoplasm of spermatogonia in both the pre- and post-pubertal stages, as well as in spermatocytes and round spermatids in the post-pubertal stage.
- GATA4-positive Sertoli cells and Leydig cells in the interstitial space showed no immune response with VASA. These cells play a vital role in supporting and regulating the function of the germ cells.
- Interestingly, the intensity of immune response was significantly stronger in pachytene spermatocytes compared to spermatogonia and round spermatids.
- Last but not least, the VASA antibody proved effective for staining individual germ cell preparations.
Conclusions
- Based on these results, the researchers concluded that VASA protein expression can be used as a reliable marker to identify spermatogonia, spermatocytes and round spermatids in the testicular tissues of stallions.
- This study’s findings could provide a new approach for observing and studying the biology of reproduction, specifically the spermatogenesis process, in equine species.
Cite This Article
APA
Kim JY, Jung HJ, Yoon MJ.
(2015).
VASA (DDX4) is a Putative Marker for Spermatogonia, Spermatocytes and Round Spermatids in Stallions.
Reprod Domest Anim, 50(6), 1032-1038.
https://doi.org/10.1111/rda.12632 Publication
Researcher Affiliations
- Department of Animal Science, Kyungpook National University, Sangju, Korea.
- Department of Animal and Biotechnology Science, Kyungpook National University, Sangju, Korea.
- Department of Animal and Biotechnology Science, Kyungpook National University, Sangju, Korea.
- Department of Horse, Companion and Wild Animal Science, Kyungpook National University, Sangju, Korea.
MeSH Terms
- Animals
- Biomarkers / metabolism
- Blotting, Western
- DEAD-box RNA Helicases / metabolism
- Horses
- Male
- Reproduction
- Sertoli Cells / metabolism
- Spermatogenesis / physiology
- Spermatozoa / cytology
- Testis / physiology
Citations
This article has been cited 11 times.- Chang JH, Chou CH, Wu JC, Liao KM, Luo WJ, Hsu WL, Chen XR, Yu SL, Pan SH, Yang PC, Su KY. LCRMP-1 is required for spermatogenesis and stabilises spermatid F-actin organization via the PI3K-Akt pathway.. Commun Biol 2023 Apr 10;6(1):389.
- Navarro S, Crespo D, Schulz RW, Ge W, Rotllant J, Cerdá-Reverter JM, Rocha A. Role of the Melanocortin System in Gonadal Steroidogenesis of Zebrafish.. Animals (Basel) 2022 Oct 12;12(20).
- Schröder MAM, Sweep FCGJ, van Herwaarden AE, Mitchell RT, Eliveld J, van Pelt AMM, Rowan AE, Korbie D, Stikkelbroeck NMML, Claahsen-van der Grinten HL, Span PN. Transcriptional comparison of testicular adrenal rest tumors with fetal and adult tissues.. Eur J Endocrinol 2022 Nov 1;187(5):607-615.
- Sakib S, Lara NLEM, Huynh BC, Dobrinski I. Organotypic Rat Testicular Organoids for the Study of Testicular Maturation and Toxicology.. Front Endocrinol (Lausanne) 2022;13:892342.
- Wang M, Ding H, Wu S, Wang M, Wei C, Wang B, Bao Z, Hu J. Vasa Is a Potential Germ Cell Marker in Leopard Coral Grouper (Plectropomus leopardus).. Genes (Basel) 2022 Jun 16;13(6).
- Chioccarelli T, Migliaccio M, Suglia A, Manfrevola F, Porreca V, Diano N, Errico S, Fasano S, Cobellis G. Characterization of Estrogenic Activity and Site-Specific Accumulation of Bisphenol-A in Epididymal Fat Pad: Interfering Effects on the Endocannabinoid System and Temporal Progression of Germ Cells.. Int J Mol Sci 2021 Mar 3;22(5).
- Maekura K, Tsukamoto S, Hamada-Kanazawa M, Takano M. Rimklb mutation causes male infertility in mice.. Sci Rep 2021 Feb 25;11(1):4604.
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- Del Vento F, Vermeulen M, Ucakar B, Poels J, des Rieux A, Wyns C. Significant Benefits of Nanoparticles Containing a Necrosis Inhibitor on Mice Testicular Tissue Autografts Outcomes.. Int J Mol Sci 2019 Nov 20;20(23).
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