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Characterization and enrichment of spermatogonial stem cells of common carp (Cyprinus carpio)

文献类型： 外文期刊

作者： Vigoya, Angel A. A. 1 ; Martinez, Emanuel R. M. 3 ; Digmayer, Melanie 3 ; Oliveira, Marcos A. de 1 ; Butzge, Arno J. 3 ; Rosa, Ivana F. 3 ; Doretto, Lucas B. 3 ; Nobrega, Rafael H. 3 ;

作者机构： 1.Aquaculture Ctr Sao Paulo State Univ, CAUNESP, BR-14884900 Jaboticabal, SP, Brazil

2.San Martin Univ Fdn FUSM, Fac Vet Med & Anim Sci, Bogota 760030, Colombia

3.Sao Paulo State Univ UNESP, Inst Biosci, Dept Struct & Funct Biol, BR-01049010 Botucatu, Brazil

4.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

5.Natl Key Lab Mariculture Biobreeding & Sustainable, Qingdao 266071, Peoples R China

关键词： Common carp(Cyprinus carpio); Stem cell markers; GFR alpha-1; POU2; Spermatogenesis depletion; Percoll gradient; Spermatogonia stem cell enrichment

期刊名称：THERIOGENOLOGY （ 影响因子：2.8； 五年影响因子：2.6 ）

ISSN： 0093-691X

年卷期： 2024 年 214 卷

页码：

收录情况： SCI

摘要： Spermatogenesis is a systematically organized process that ensures uninterrupted sperm production in which the spermatogonial stem cells (SSCs) play a crucial role. However, the existing absence of teleost-specific molecular markers for SSCs presents a notable challenge. Herein we characterized phenotypically the spermatogonial stem cells using specific molecular markers and transmission electron microscopy. Moreover, we also describe a simple method to suppress common carp spermatogenesis using the combination of Busulfan and thermo-chemical treatment, and finally, we isolate and enrich the undifferentiated spermatogonial fraction. Our results showed that C-kit, GFR alpha 1, and POU2 proteins were expressed by germ cells, meanwhile, undifferentiated spermatogonial populations preferentially expressed GFR alpha 1 and POU2. Moreover, the combination of high temperature (35 degrees C) and Busulfan (40 mg/kg/BW) effectively suppressed the spermatogenesis of common carp males. Additionally, the amh expression analysis showed differences between the control (26 degrees C) when compared to 35 degrees C with a single or two Busulfan doses, confirming that the testes were depleted by the association of Busulfan at high temperatures. In an attempt to isolate the undifferentiated spermatogonial fraction, we used the Percoll discontinuous density gradient. Thus, we successfully dissociated the carp whole testes in different cellular fractions; subsequently, we isolated and enriched the undifferentiated spermatogonial population. Therefore, our results suggest that probably both GFR alpha-1 and POU2 are highly conserved factors expressed in common carp germinative epithelium and that these molecules were well conserved along the evolutionary process. Furthermore, the enriched undifferentiated spermatogonial population developed here can be used in further germ cell transplantation experiments to preserve and propagate valued and endangered fish species.

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