Autophagy in the fat body cells of the cave cricket Troglophilus neglectus Krauss, 1878 (Rhaphidophoridae, Saltatoria) during overwintering
Autophagy in the fat body cells of the cave cricket Troglophilus neglectus Krauss, 1878 (Rhaphidophoridae, Saltatoria) during overwintering
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DOI：
10.1007/s00709-015-0824-3
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发表时间：
2016-03-01
期刊：
影响因子：
2.9
通讯作者：
Novak, Tone
中科院分区：
文献类型：
--
作者：
Lipovsek, Saska;Novak, Tone
洞穴蟋蟀Troglophilus neglectus定期在地下水栖息地越冬4-5个月。冬季休眠是一个自然的饥饿期，为研究自然条件下的自噬提供了机会。我们的目的是评估在三个时间范围内的脂肪体的脂肪细胞和尿细胞的自噬活性：直接越冬前，在中间的休眠，并在其结束。为此，我们对洞穴中的个体进行了采样。通过透射电子显微镜（TEM）和免疫荧光显微镜（IFM）的自噬体的丰度的细胞超微结构进行了研究，应用广泛使用的，特异性免疫标记标记微管相关蛋白1轻链3（LC 3）。在越冬前，透射电镜显示脂肪细胞中自噬体和残留体很少，而尿细胞中没有。相应地，IFM显示出非常有限的反应或没有反应。在越冬中期和末期，在两种细胞类型中，吞噬细胞，自噬体，自溶酶体，和残留体的TEM鉴定，而LC 3免疫标记检测自噬体显示出明显的阳性反应。两种方法都显示，在任何时间范围内，两性之间都没有显著差异。自噬活性在越冬休眠前最低，到越冬结束时逐渐增强，可能是由于蛋白质颗粒中的储备蛋白不完全由所需氨基酸组成。我们的结论是，在T。自噬是对饥饿的实质性反应，并通过为细胞提供营养来支持冬季休眠期间的稳态过程。
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.