Ozkaraca Mustafa, Ozdemir Selcuk, Comakli Selim, Timurkan Mehmet Ozkan
Department of Pathology, Faculty of Veterinary Medicine, Cumhuriyet University, Sivas, Turkey.
Department of Genetic, Faculty of Veterinary Medicine, Ataturk University, Erzurum, Turkey.
Vet Med (Praha). 2022 Jan 15;67(1):1-12. doi: 10.17221/221/2020-VETMED. eCollection 2022 Jan.
The aim of this study was to investigate the activity of apoptosis and autophagy in animals (cows, horses, donkeys, dogs and cats) naturally infected with rabies by using immunohistochemistry, immunofluorescence, and qPCR. The mRNA transcript levels of caspase-3, Bax, Bcl2 and LC3B were determined with qPCR. Caspase-3 and AIF immunopositivity were not observed in the immunohistochemical and immunofluorescence staining, whereas LC3B immunopositivity was determined intensively in the infected animals compared to the control groups. LC3B immunopositivity was detected in the cytoplasm of the Purkinje cells in the cerebellum of the cows, horses and donkeys, and also in the cytoplasm of the neurons in the cornu ammonis of the dogs and cats. While the expression levels of caspase-3 and Bax were downregulated, the Bcl2 expression was up-regulated in the infected animals compared to the uninfected animals. In addition, the LC3B levels were found to be significantly higher in the infected animals. To the best of our knowledge, this work represents the first report of neuronal death in the central nervous system by autophagy, rather than by caspase-dependent or AIF-containing caspase-independent apoptosis.
本研究的目的是通过免疫组织化学、免疫荧光和定量聚合酶链反应（qPCR），调查自然感染狂犬病的动物（牛、马、驴、狗和猫）中凋亡和自噬的活性。用qPCR测定半胱天冬酶-3（caspase-3）、Bax、Bcl2和微管相关蛋白1轻链3β（LC3B）的mRNA转录水平。在免疫组织化学和免疫荧光染色中未观察到半胱天冬酶-3和凋亡诱导因子（AIF）的免疫阳性，而与对照组相比，在感染动物中强烈检测到LC3B免疫阳性。在牛、马和驴小脑的浦肯野细胞胞质中检测到LC3B免疫阳性，在狗和猫海马的神经元胞质中也检测到LC3B免疫阳性。与未感染动物相比，感染动物中半胱天冬酶-3和Bax的表达水平下调，而Bcl2表达上调。此外，发现感染动物中LC3B水平显著更高。据我们所知，这项工作代表了中枢神经系统中神经元死亡是通过自噬而非依赖半胱天冬酶或含AIF的非依赖半胱天冬酶凋亡的首次报道。
