One thousand five hundred cachara or tiger shovelnose catfish Pseudoplatystoma fasciatum, obtaine... more One thousand five hundred cachara or tiger shovelnose catfish Pseudoplatystoma fasciatum, obtained from induced reproduction, were used to determine the onset of ovarian differentiation and development and to record the main characteristics of this process. Samples were collected from 0 to 240 days post-fertilization (dpf) and the results classified into stages I-XII. Ovarian formation was histologically detected for the first time when juveniles measured mean ± s.d. 51·5 ± 8·3 mm total length (LT ) at 39-45 dpf (stages I-V), with intense somatic cell proliferation originating in the ovarian cavity. Both LT and age of fish had a positive correlation (P < 0·001) with ovarian differentiation, but LT showed a greater correlation (r(2) = 0·95) than age (r(2) = 0·85), especially during the initial stages of development. From stages VI to VII, the ovarian cavity was enlarged and undifferentiated oogonia were present. At stage VIII, small projections formed in the ovarian stroma towar...
5 Gabriela Brambila-Souza 1, Constantinos Chrysanthos Mylonas 2, Paulo 6 Henrique de Mello 1, Raf... more 5 Gabriela Brambila-Souza 1, Constantinos Chrysanthos Mylonas 2, Paulo 6 Henrique de Mello 1, Rafael Yutaka Kuradomi 3, Sergio Ricardo Batlouni 1, 7 Carlos Eduardo Tolussi 4, Renata Guimarães Moreira 5*. 8 9 1 UNESPUniversidade Estadual Paulista Centro de Aquicultura (CAUNESP), Jaboticabal, 10 SP, Brazil. 11 2 Institute of Marine Biology, Biotechnology and Aquaculture, Hellenic Center for Marine 12 Research, Heraklion Crete, Greece. 13 3 – Universidade Federal do Amazonas, Manaus, AM, Brazil. 14 4 – Universidade Anhembi Morumbi, São Paulo, SP, Brazil. 15 5 Instituto de Biociências –Universidade de São Paulo (USP), SP, Brazil. 16 *Corresponding author: renatagm@ib.usp.br 17
The objective of this study was to establish a protocol for the characterization, isolation, and ... more The objective of this study was to establish a protocol for the characterization, isolation, and culture of type A spermatogonia using specific molecular markers for these cells in fish. To this end, adult Prochilodus lineatus testes were collected and digested enzymatically and the resulting testicular suspension was separated using a discontinuous Percoll gradient, followed by differential plating. The cell cultures obtained were monitored for 15 days and analyzed using the immunofluorescence method with anti‐Vasa, anti‐GFRα1, and anti‐OCT4 antibodies. Spermatogonial enrichment was also performed using flow cytometry. Although discontinuous Percoll gradient centrifugation followed by differential plating enabled the removal of differentiated germ cells and somatic cells, enriching the pool of type A spermatogonia, the enrichment of type A spermatogonia through flow cytometry of samples without Percoll proved to be more efficient. Prominent cell agglomerates that were characterized according to different stem cell markers as type A spermatogonia were observed during the 15 days of the cell culture. The use of immunoperoxidase and western blot analysis methods confirmed the specificity of the markers for type A spermatogonia of P. lineatus. When combined with specific cell culture conditions, the positive characterization of these molecular markers clarified certain aspects of spermatogonial regulation, such as survival and proliferation. Finally, understanding the regulation of the in vitro germ cell maintenance process may contribute to the enhancement of in vivo and in vitro reproduction techniques of endangered or aquaculture fish species.
In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected d... more In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 μm (82.5% with ...
Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêm... more Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêmeas “jovens” (FJs) de P. mesopotamicus, visando melhorar o processo de seleção de peixes para a indução hormonal. Avaliamos também a acurácia do uso de características corporais externas para a seleção de fêmeas para indução hormonal. Para isso, 60 FVs e 60 FJs (10 e quatro anos, respectivamente) foram submetidas a sete amostragens durante dois ciclos reprodutivos. Em cada amostra, as fêmeas (cinco a 10 por tratamento) foram capturadas para avaliação das concentrações plasmáticas de esteroides gonadais e composição dos ovários (biópsias e estereologia). As fêmeas foram também submetidas à uma avaliação externa relativa à espessura abdominal, o grau de rigidez abdominal e o aspecto da papila urogenital. Exceto pela ocorrência de concentrações reduzidas de 17α-hidroxiprogesterona nas FVs, durante a época de desova, observamos que FVs e FJs , mantidas em cativeiro, apresentam ciclos reprodu...
One thousand five hundred cachara or tiger shovelnose catfish Pseudoplatystoma fasciatum, obtaine... more One thousand five hundred cachara or tiger shovelnose catfish Pseudoplatystoma fasciatum, obtained from induced reproduction, were used to determine the onset of ovarian differentiation and development and to record the main characteristics of this process. Samples were collected from 0 to 240 days post-fertilization (dpf) and the results classified into stages I-XII. Ovarian formation was histologically detected for the first time when juveniles measured mean ± s.d. 51·5 ± 8·3 mm total length (LT ) at 39-45 dpf (stages I-V), with intense somatic cell proliferation originating in the ovarian cavity. Both LT and age of fish had a positive correlation (P < 0·001) with ovarian differentiation, but LT showed a greater correlation (r(2) = 0·95) than age (r(2) = 0·85), especially during the initial stages of development. From stages VI to VII, the ovarian cavity was enlarged and undifferentiated oogonia were present. At stage VIII, small projections formed in the ovarian stroma towar...
5 Gabriela Brambila-Souza 1, Constantinos Chrysanthos Mylonas 2, Paulo 6 Henrique de Mello 1, Raf... more 5 Gabriela Brambila-Souza 1, Constantinos Chrysanthos Mylonas 2, Paulo 6 Henrique de Mello 1, Rafael Yutaka Kuradomi 3, Sergio Ricardo Batlouni 1, 7 Carlos Eduardo Tolussi 4, Renata Guimarães Moreira 5*. 8 9 1 UNESPUniversidade Estadual Paulista Centro de Aquicultura (CAUNESP), Jaboticabal, 10 SP, Brazil. 11 2 Institute of Marine Biology, Biotechnology and Aquaculture, Hellenic Center for Marine 12 Research, Heraklion Crete, Greece. 13 3 – Universidade Federal do Amazonas, Manaus, AM, Brazil. 14 4 – Universidade Anhembi Morumbi, São Paulo, SP, Brazil. 15 5 Instituto de Biociências –Universidade de São Paulo (USP), SP, Brazil. 16 *Corresponding author: renatagm@ib.usp.br 17
The objective of this study was to establish a protocol for the characterization, isolation, and ... more The objective of this study was to establish a protocol for the characterization, isolation, and culture of type A spermatogonia using specific molecular markers for these cells in fish. To this end, adult Prochilodus lineatus testes were collected and digested enzymatically and the resulting testicular suspension was separated using a discontinuous Percoll gradient, followed by differential plating. The cell cultures obtained were monitored for 15 days and analyzed using the immunofluorescence method with anti‐Vasa, anti‐GFRα1, and anti‐OCT4 antibodies. Spermatogonial enrichment was also performed using flow cytometry. Although discontinuous Percoll gradient centrifugation followed by differential plating enabled the removal of differentiated germ cells and somatic cells, enriching the pool of type A spermatogonia, the enrichment of type A spermatogonia through flow cytometry of samples without Percoll proved to be more efficient. Prominent cell agglomerates that were characterized according to different stem cell markers as type A spermatogonia were observed during the 15 days of the cell culture. The use of immunoperoxidase and western blot analysis methods confirmed the specificity of the markers for type A spermatogonia of P. lineatus. When combined with specific cell culture conditions, the positive characterization of these molecular markers clarified certain aspects of spermatogonial regulation, such as survival and proliferation. Finally, understanding the regulation of the in vitro germ cell maintenance process may contribute to the enhancement of in vivo and in vitro reproduction techniques of endangered or aquaculture fish species.
In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected d... more In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 μm (82.5% with ...
Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêm... more Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêmeas “jovens” (FJs) de P. mesopotamicus, visando melhorar o processo de seleção de peixes para a indução hormonal. Avaliamos também a acurácia do uso de características corporais externas para a seleção de fêmeas para indução hormonal. Para isso, 60 FVs e 60 FJs (10 e quatro anos, respectivamente) foram submetidas a sete amostragens durante dois ciclos reprodutivos. Em cada amostra, as fêmeas (cinco a 10 por tratamento) foram capturadas para avaliação das concentrações plasmáticas de esteroides gonadais e composição dos ovários (biópsias e estereologia). As fêmeas foram também submetidas à uma avaliação externa relativa à espessura abdominal, o grau de rigidez abdominal e o aspecto da papila urogenital. Exceto pela ocorrência de concentrações reduzidas de 17α-hidroxiprogesterona nas FVs, durante a época de desova, observamos que FVs e FJs , mantidas em cativeiro, apresentam ciclos reprodu...
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