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Autophagy and apoptosis interplay during follicular atresia in fish ovary: a morphological and immunocytochemical study

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Abstract

Follicular atresia in fish ovary provides an interesting model for studying autophagy and apoptosis. In order to improve knowledge of the mechanisms regulating ovarian regression, we investigated the immunolocalisation of various proteins involved in the complex network of autophagy and apoptosis. Females of three species of freshwater fish maintained in captivity were sampled after the reproductive period and the main events of follicular atresia were assessed by histology: splits in the zona radiata, yolk degradation and reabsorption, hypertrophy of the follicular cells, accumulation of autophagic vacuoles, closing of the follicular lumen and thickening of the theca. The interplay of apoptosis and autophagy was analysed by TUNEL in situ and by immunocytochemistry for caspase-3, bax, bcl-2, beclin-1 and cathepsin-D. During early and advanced stages of follicular regression, the actin cytoskeleton was well developed and labelling for bcl-2 and cathepsin-D were pronounced in the follicular cells at a stage when they were intensively involved in yolk phagocytosis. Immunofluorescence for beclin-1 was prevalent in the follicular cells, punctate labelling often surrounding autophagic vacuoles during the advanced stage of follicular regression, a critical step towards cell death. TUNEL-positive reaction and immunostaining for bax and caspase-3 demonstrated the participation of apoptosis in late follicular regression. Overall, this study provides evidence that autophagic and apoptotic proteins are activated in a coordinated fashion depending on the stage of follicular regression, with interplay between autophagy and apoptosis being essential in determining the fate of the cell during follicular atresia in fish ovary.

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Acknowledgements

We thank the Estação de Hidrobiologia e Piscicultura de Três Marias, Companhia de Desenvolvimento dos Vales do São Francisco e Parnaíba, for technical assistance during fish collections. We are also grateful to Dr. Paulo Pimenta and colleagues of the Centro de Pesquisas René Rachou for technical assistance with image collection by confocal laser microscopy and to Steve Latham for valuable suggestions with regard to the English language.

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Authors and Affiliations

  1. Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, C.P. 486, 30161-970, Belo Horizonte, Minas Gerais, Brasil

    R. D. V. S. Morais, R. G. Thomé, F. S. Lemos & E. Rizzo

  2. Programa de Pós-Graduação em Zoologia de Vertebrados, Pontifícia Universidade Católica de Minas Gerais, PUC Minas, Belo Horizonte, Minas Gerais, Brasil

    N. Bazzoli

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  1. R. D. V. S. Morais
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  2. R. G. Thomé
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  3. F. S. Lemos
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  4. N. Bazzoli
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  5. E. Rizzo
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Correspondence to E. Rizzo.

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This study was supported by grants from Brazilian Research Foundations: FAPEMIG, CNPq and CAPES.

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Morais, R.D.V.S., Thomé, R.G., Lemos, F.S. et al. Autophagy and apoptosis interplay during follicular atresia in fish ovary: a morphological and immunocytochemical study. Cell Tissue Res 347, 467–478 (2012). https://doi.org/10.1007/s00441-012-1327-6

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  • DOI: https://doi.org/10.1007/s00441-012-1327-6

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