Immunohistochemical localisation of Glutamate ammonia ligase (GLUL) in cerebrum in different clinical forms of canine rabies

P. Nikhithasree, C. Divya, M.L. Arya Nair, K. Krithiga, K.S. Prasanna, Lali F. Anand, John Bernet Johnson
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Abstract

Rabies, one of the earliest known diseases reported in the history of mankind, is a fatal acute viral disease of the central nervous system. Despite numerous studies on etiology of rabies, its exact mechanism of neuropathogenesis remains unidentified. Glutamate ammonia ligase (GLUL) is a protein present in brain which play an important role in the pathogenesis of many neurological conditions. GLUL was identified to be over expressed in brain of human rabies. Cerebral cortex is the part of brain primarily responsible for coordination of movements and behaviour in animals and humans. Hence the work was designed to study the immunohistochemical localisation of GLUL in cerebrum in rabid carcasses in order to elucidate its role in the pathology of this deadly viral infection. The carcasses of 219 rabies suspected dogs that were brought to the Department of Veterinary Pathology, CVAS, Mannuthy, for necropsy between January 2021 and August 2022 formed the materials for the study. One hundred and thirty-three cases were found positive for rabies using the gold standard test for rabies - Direct Fluorescent Antibody Test (dFAT) which was further confirmed by polymerase chain reaction targeting N gene with amplicon size 533 bp. Among these 133 confirmed cases, 30 selected samples (frontal, occipital, temporal, parietal lobes of cerebrum) were further processed for histopathological and immunohistochemical (IHC) studies. The IHC signals of GLUL obtained were compared with the clinical form of rabies. Immunohistochemical staining revealed localisation of GLUL in glial cells with different staining intensity in encephalitic and paralytic form of rabies. The study concluded that the IHC signals for GLUL were stronger for furious form of rabies and weak for dumb form of the disease.
不同临床形式犬狂犬病大脑谷氨酸氨连接酶(GLUL)的免疫组织化学定位
狂犬病是人类历史上最早报道的已知疾病之一,是一种致命的中枢神经系统急性病毒性疾病。尽管对狂犬病的病因学进行了大量的研究,但其确切的神经发病机制仍未确定。谷氨酸氨连接酶(GLUL)是存在于大脑中的一种蛋白,在许多神经系统疾病的发病机制中起重要作用。经鉴定,GLUL在人狂犬病脑组织中过表达。大脑皮层是大脑的一部分,主要负责协调动物和人类的运动和行为。因此,这项工作旨在研究狂犬病尸体大脑中GLUL的免疫组织化学定位,以阐明其在这种致命病毒感染的病理中的作用。2021年1月至2022年8月期间,219只疑似狂犬病狗的尸体被带到曼努西兽医病理学部进行尸检,形成了该研究的材料。采用狂犬病金标准检测方法-直接荧光抗体试验(dFAT)检测出133例狂犬病阳性,并采用靶向扩增子大小为533 bp的N基因聚合酶链反应进一步证实。在这133例确诊病例中,选择30个样本(大脑额叶、枕叶、颞叶、顶叶)进行进一步的组织病理学和免疫组化(IHC)研究。将获得的GLUL免疫组化信号与狂犬病临床表现进行比较。免疫组化染色显示脑瘫型和麻痹型狂犬不同染色强度的胶质细胞中有GLUL的定位。研究结果表明,狂犬型GLUL的IHC信号较强,而非狂犬型GLUL的IHC信号较弱。
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