ITRAQ‐based proteomic analysis of the hippocampus of pentylenetetrazole‐kindled epileptic rats. Issue 2 (4th January 2021)

• Record Type: Journal Article
• Authors: Xu, Weiye; Zhang, Siyuan; Feng, Yanyan; Zhang, Chen; Xiao, Yeqing; Tian, Fafa
• Is Part Of: International journal of developmental neuroscience. Volume 81:Issue 2(2021:Apr.)
• Page Start: 125
• Page End: 141
• Publication Date: 2021-01-04
• Subjects: epileptogenesis -- hippocampus -- pentylenetetrazole‐kindled model -- proteomics; Developmental neurobiology -- Periodicals; Neurology -- Periodicals; Neurologie du développement -- Périodiques; Developmental neurobiology; Periodicals; 612.8
• DOI: 10.1002/jdn.10082
• Languages: English
• ISSNs: 0736-5748

Abstract

Abstract: Epilepsy can severely affect the quality of life of patients, who are often at higher risk of mortality. However, the molecular mechanisms and pathogenesis underlying epileptogenesis are poorly understood. In this study, we performed a proteomic analysis of the hippocampus in pentylenetetrazole (PTZ)‐kindled epileptic rats to explore the molecular mechanisms of epileptogenesis. We established an epileptic model in Sprague Dawley rats by injecting PTZ intraperitoneally and applied isobaric tags for relative and absolute quantification (iTRAQ) technology integrated with liquid chromatography–tandem mass spectrometry (LC–MS/MS) to identify differentially expressed proteins (DEPs) in the hippocampus. A total of 99 proteins, comprising 93 upregulated and 6 downregulated proteins, were identified based on a fold change >1.2 (or <0.83) and a p ‐value < .05. A further bioinformatics analysis suggested that the candidate proteins were mainly involved in the ubiquitin ligase complex or metabolite homeostasis or acted as intrinsic components of the membrane. A Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway enrichment analysis identified a series of representative pathological pathways, including the calcium signaling pathway, neuroactive ligand‐receptor interaction pathway, and the NF‐kappa B pathway. The mass spectrometry results were further confirmed by assessing five representative proteins (Akt1, Syvn1, Amfr, Lamb1, and Cox17) using western blotting and immunohistochemistry. These results may help to reveal the molecular mechanisms underlying epileptogenesis and provide new directions or targets for epilepsy research. Abstract : The present study aims to investigate the proteomic profile of the brain in an epileptic rat model. A total of 99 proteins were identified as differentially expressed proteins according to a threshold of a 1.2‐fold change with a p ‐value < .05. A bioinformatics analysis showed that the functions of the 99 differential proteins were mainly concentrated in ubiquitin‐protein ligase, synaptic transmission, the calcium signaling pathway, etc. Five representative proteins (Akt1, Syvn1, Amfr, Lamb1, and Cox17) were then further confirmed by western blotting and immunohistochemistry.