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

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.