Preliminary study on testicular germ cell isolation and transplantation in an endangered endemic species Brycon orbignyanus (Characiformes: Characidae)

被引:11
作者
de Siqueira-Silva, Diogenes Henrique [1 ,2 ,3 ]
dos Santos Silva, Amanda Pereira [2 ]
da Silva Costa, Raphael [1 ,2 ]
Senhorini, Jose Augusto [4 ]
Ninhaus-Silveira, Alexandre [2 ]
Verissimo-Silveira, Rosicleire [1 ,2 ]
机构
[1] UNESP Univ Estadual Paulista, Dept Biol & Anim Sci, LINEO Lab Ictiol Neotrop, Campus Ilha Solteira,Ave Brasil Ctr 59, BR-15385000 Ilha Solteira, SP, Brazil
[2] UNESP Univ Estadual Paulista, Postgrad Program Anim Biol, Campus Sao Jose Do Rio Preto, BR-15504000 Sao Jose Do Rio Preto, SP, Brazil
[3] UNESP Univ Estadual Paulista, Dept Biol & Zootecnia, LINEO Lab Ictiol Neotrop, Campus Ilha Solteira,Rua Moncao 226, Sao Paulo, Brazil
[4] Chico Mendes Inst Biodivers Conservat, CEPTA ICMBIO Natl Ctr Res & Conservat Continenta, Rodovia Pref Euberto Nemesio Pereira Godoy, BR-13630970 Pirassununga, SP, Brazil
关键词
Fish chimera; Germ cell technology; Spermatogonial proliferation; Surrogate production; LINE CHIMERAS; FISH; PROLIFERATION; SPERMATOGONIA; COLONIZATION; GENERATION; SURVIVAL;
D O I
10.1007/s10695-019-00631-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We aimed to develop a simplified protocol for transplantation of Brycon orbignyanus spermatogonial stem cells (SSCs) into Astyanax altiparanae testes. Brycon orbignyanus testes were enzymatically digested and SSC purified by a discontinuous density gradient. Endogenous spermatogenesis was suppressed in A. altiparanae using busulfan or by incubation at 35 degrees C water, and SSCs from B. orbignyanus labeled with PKH26 were injected into their testes via the urogenital papilla. Twenty-two hours post-transplantation, labeled spermatogonia were observed in A. altiparanae tubular lumen. After 7 days, spermatogonia proliferated in the epithelium, and 21 days post-transplantation, sperm was observed in the lumen. Of surviving host fish, nearly 67% of those treated with busulfan and 85% of those held in warm water showed labeled cells in host germinal epithelium. The present study standardized, by a simple and accessible method, germ cell transplantation between sexually mature Characiformes fish species. This is the first report of xenogenic SSC transplantation in this fish order.
引用
收藏
页码:767 / 776
页数:10
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