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

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Abstract

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 °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.

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Acknowledgments

The authors are deeply grateful to Douglas de Castro Ribeiro for help constructing the aquarium system and to Usina Hidrelétrica Engenheiro Souza Dias (Jupiá-CESP) and Centro Nacional de Pesquisa e Conservação da Biota Aquática Continental (CEPTA) for providing the fish used in this study.

Funding

This study was supported by Fundação de Apoio à Pesquisa de São Paulo (FAPESP; Process number: 2013/24218-5) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; grant number: 160-380/2011).

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Authors and Affiliations

  1. Campus de Ilha Solteira, Departament of Biology and Animal Science, L.I.NEO – Laboratório de Ictiologia Neotropical, UNESP – Univ. Estadual Paulista, Avenida Brasil Centro, 56, Ilha Solteira, Sao Paulo, 15385-000, Brazil

    Diógenes Henrique de Siqueira-Silva, Alexandre Ninhaus-Silveira & Rosicleire Veríssimo-Silveira

  2. Campus de São José do Rio Preto, Post-Graduation Program in Animal Biology, UNESP – Univ. Estadual Paulista, Rua Cristovão Colombo, Jardim Nazareth, 2265, São José do Rio Preto, Sao Paulo, 15504-000, Brazil

    Diógenes Henrique de Siqueira-Silva, Amanda Pereira dos Santos Silva, Raphael da Silva Costa, Alexandre Ninhaus-Silveira & Rosicleire Veríssimo-Silveira

  3. Campus de Ilha Solteira, Departamento de Biologia e Zootecnia, L.I.NEO - Laboratório de Ictiologia Neotropical, UNESP –Univ. Estadual Paulista, Rua Monção, 226, Zona Norte, Sao Paulo, Brazil

    Diógenes Henrique de Siqueira-Silva

  4. CEPTA-ICMBIO - National Center for Research and Conservation of Continental Fish, Chico Mendes Institute of Biodiversity Conservation, Rodovia Pref. Euberto Nemesio Pereira de Godoy, Pirassununga, Sao Paulo, 13630-970, Brazil

    José Augusto Senhorini

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  1. Diógenes Henrique de Siqueira-Silva
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  2. Amanda Pereira dos Santos Silva
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  3. Raphael da Silva Costa
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  4. José Augusto Senhorini
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  5. Alexandre Ninhaus-Silveira
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  6. Rosicleire Veríssimo-Silveira
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Corresponding author

Correspondence to Diógenes Henrique de Siqueira-Silva.

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Experimental procedures were conducted in accordance with the guidance for care and use of animals in the laboratory of Universidade Estadual Paulista (UNESP-Ilha Solteira). The Committee of Ethics and Research of UNESP approved the protocol 006/2012/CONCEA.

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de Siqueira-Silva, D.H., dos Santos Silva, A.P., da Silva Costa, R. et al. Preliminary study on testicular germ cell isolation and transplantation in an endangered endemic species Brycon orbignyanus (Characiformes: Characidae). Fish Physiol Biochem 47, 767–776 (2021). https://doi.org/10.1007/s10695-019-00631-8

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  • DOI: https://doi.org/10.1007/s10695-019-00631-8

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