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Autophagy in the fat body cells of the cave cricket Troglophilus neglectus Krauss, 1878 (Rhaphidophoridae, Saltatoria) during overwintering

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Abstract

The cave cricket Troglophilus neglectus regularly overwinters for 4–5 months in hypogean habitats. Winter dormancy is a natural starvation period, providing the opportunity to study autophagy under natural conditions. We aimed to evaluate the autophagic activity in adipocytes and urocytes of the fat body in three time frames: directly before overwintering, in the middle of dormancy, and at its end. For this purpose, we sampled individuals in caves. The cell ultrastructure was studied by transmission electron microscopy (TEM) and the abundance of autophagosomes by immunofluorescence microscopy (IFM), applying the widely used, specific immunolabeling marker microtubule-associated protein 1 light chain 3 (LC3). Before overwintering, TEM revealed scarce autophagosomes and residual bodies in the adipocytes and none in the urocytes. Congruently, IFM showed a very limited or no reaction. In the middle and at the end of overwintering, in both cell types, phagophores, autophagosomes, autolysosomes, and residual bodies were identified by TEM, while LC3 immunolabeling for detecting autophagosomes showed a conspicuous positive reaction. Both methods revealed that there were no significant differences between the sexes in any time frame. Minimal autophagic activity was detected before the winter dormancy, and it gradually intensified till the end of overwintering, probably because reserve proteins in protein granula are not composed of all the required amino acids. We conclude that in T. neglectus, autophagy is a substantial response to starvation and supports homeostatic processes during winter dormancy by supplying cells with nutrients.

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Acknowledgments

We thank Elisabeth Bock, Rudi Schmied (Medical University Graz), and Rudi Mlakar (Faculty of Medicine, University of Maribor) for their excellent technical assistance, Franc Janžekovič for the statistical analysis of the data, and Michelle Gadpaille for the linguistic improvements to the manuscript. We are grateful to two anonymous reviewers for their insightful comments and suggestions. This study was partly supported by the Biodiversity Research Programme, Grant P1-0078.

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Authors and Affiliations

  1. Faculty of Medicine, University of Maribor, Taborska ulica 8, 2000, Maribor, Slovenia

    Saška Lipovšek

  2. Department of Biology, Faculty of Natural Sciences and Mathematics, University of Maribor, Koroška 160, 2000, Maribor, Slovenia

    Saška Lipovšek & Tone Novak

  3. Faculty of Chemistry and Chemical Engineering, University of Maribor, Smetanova ulica 17, 2000, Maribor, Slovenia

    Saška Lipovšek

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  1. Saška Lipovšek
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  2. Tone Novak
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Correspondence to Saška Lipovšek.

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Lipovšek, S., Novak, T. Autophagy in the fat body cells of the cave cricket Troglophilus neglectus Krauss, 1878 (Rhaphidophoridae, Saltatoria) during overwintering. Protoplasma 253, 457–466 (2016). https://doi.org/10.1007/s00709-015-0824-3

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