Recent studies seem to indicate that apoptosis and autophagy can act cooperatively in fish ovaries in order to achieve more effective ovarian regression after spawning. Considering the importance of tissue remodeling in ovarian functioning, we sought to morphologically characterize the involution processes of follicular atresia and post-ovulatory complexes using Astyanax altiparanae as an experimental model, and to determine the location of proteins involved in apoptosis and autophagy throughout this process. Fifteen females were collected after reproductive management. Fragments of the left ovaries were removed, fixed, and prepared for light microscopy and immunofluorescence analyses. The main characteristics of the involution processes were found to be consistent with previous descriptions. However, there were certain morphological peculiarities that do not appear to have been described for any other species thus far. These peculiarities may be related to the focus of this study on a single species, which allows for a more detailed investigation into morphological changes than studies on multiple species. Autophagy was also found to precede apoptosis in both involution processes in A. altiparanae. This may be related to the energy recycling process required before the removal of degenerated follicular cells by apoptosis. Thus, these results support the idea that there is crosstalk between autophagy and apoptosis pathways in ovarian involution processes, as well as the idea that the cell death pathways of these processes are conserved between teleost species with external fertilization.
Captive reproduction is one of the problems faced in aquaculture requiring the manipulation of environmental factors and/ or hormonal treatment. Thus, we seek to verify experimentally which gonadal changes were present in mature individuals of Astyanax altiparanae arising from decreased water level. Collections were made every four hours, initiated four hours before and finished 28 hours after stimulation, at the Fish Farming Station of Companhia Energética de São Paulo, São Paulo, Brazil. The gonads were analyzed by light microscopy. The females had ovaries in the spawning capable phase until 12h; in 16h, in a more advanced stage of spawning capable phase; and, from 20h, in the regressing phase. Males had testes in the spawning capable phase until 8h; in 12h, in a more advanced stage of spawning capable phase; and, from 16h, the return to the spawning capable phase. The morphological description was corroborated by the proportion of cell classes. Females presented variation on the gonadosomatic index, but it was not found an emptying of the gonad for neither sex. The process of inducing reproduction with water level drawdown was considered satisfactory, since both sexes presented a reduction in the number of mature gametes at the end of the sample period.Reprodução em cativeiro é um dos problemas enfrentados na aquicultura exigindo a manipulação de fatores ambientais e/ ou tratamento hormonal. Assim, verificou-se experimentalmente as alterações gonadais presentes em indivíduos adultos de Astyanax altiparanae decorrentes da diminuição do nível da água. As coletas foram realizadas a cada quatro horas, iniciadas quatro horas antes e encerradas 28 horas após a estimulação, na Estação de Piscicultura da Companhia Energética de São Paulo, São Paulo, Brasil. As gônadas foram analisadas por microscopia de luz. As fêmeas apresentaram ovários na fase de apto à reprodução até 12h; em 16h, um estágio mais avançado desta fase; e, a partir das 20h, na fase de regressão. Os machos tiveram os testículos em fase de apto à reprodução até 8h; em 12h, um estágio mais avançado desta fase; e, a partir das 16h, o retorno à fase de apto à reprodução. A descrição morfológica foi corroborada pela proporção das classes celulares. As fêmeas apresentaram variação no índice gonadossomático, mas não foi estabelecido o esvaziamento das gônadas para nenhum dos sexos. O processo utilizado para a indução à reprodução foi considerado satisfatório, uma vez que ambos os sexos apresentaram redução no número de gametas maduros no final do período de amostragem.
Astyanax altiparanae is a Brazilian species of substantial commercial, environmental and scientific importance; however, existing studies on its reproduction do not seem to provide enough details. In light of the increasing use of this species in fish farming and the need for basic studies for the development of new production technologies, we describe the structural and ultrastructural characteristics of the ovaries of A. altiparanae, and characterize the species' reproductive cycle. Females were collected monthly from March 2013 to February 2014, and reproductive management began in October 2013. The ovaries were removed, fixed in Karnovsky's fixative, and prepared for light microscopy, transmission electron microscopy and immunohistochemistry anti-PCNA. These techniques enabled us to characterize the ovaries, the germ cells, and the somatic cells in detail, as well as their changes over time. The reproductive cycle was characterized based on the monthly variation in the gonadosomatic ratio, the proportion of germ cells, and the rate of oogonium proliferation. The macroscopic analysis of the ovaries suggests that the vascularization pattern and color of the ovaries vary according to development. There are new types of analyses that can be applied even in the fish farming industry, such as a comparison between ovaries staining and weight or the frequency distribution of these colors throughout the year. This study also provides details on microscopic characteristics that have never before been reported for species of Astyanax, such as the presence of annulate lamellae in oogonia, the development of the zona pellucida from oocytes in the one-nucleolus step, and the development of the micropylar apparatus in oocytes in the cortical alveolar step. When the reproductive cycle was analyzed, this species was found to have a long period of spawning, with a reproductive peak from October to February and multiple spawning events, confirming the period already described in the literature. Variations in reproductive periods and the ability to reproduce in lentic environments suggest that A. altiparanae has the ability to respond quickly to environmental changes and exhibits high reproductive flexibility. All of these characteristics confirm the great potential of this species in the fish farming industry.
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