Nuclear omics of mouse forebrain excitatory neurons in pilocarpine-induced seizures

• Main Author: Soon, Hui Rong
• Other Authors: Chng Toh Hean
• Format: Thesis-Doctor of Philosophy
• Language: English
• Published: Nanyang Technological University 2022
• Subjects: Science::Biological sciences::Human anatomy and physiology::Neurobiology; Science::Biological sciences::Molecular biology
• Online Access: https://hdl.handle.net/10356/163613
• Institution: Nanyang Technological University

An epileptic seizure generates massive neuronal activity in the brain which induces de novo transcription and translation, bringing about cellular changes which may contribute to epileptogenesis. To understand the cell-type-specific molecular changes brought about by seizures, I isolated nuclei from forebrain excitatory neurons of a pilocarpine-induced transgenic mouse and profiled the nuclear transcriptome and proteome. My nuclear transcriptome analyses show that expression of genes related to MAPK signalling are upregulated while a major class of C2H2-type zinc finger proteins, including those containing KRAB domains, are downregulated in seizure-induced animals. In parallel, nuclear proteome analyses uncovered AP-1 complex components and post-translational modifications such as SUMOylation and ubiquitin-like conjugation to be enriched in the seizure group. Finally, I show that changes in nuclear TEF levels are activity-dependent in forebrain excitatory neurons of seizure-induced mice, as well as in cultured hippocampal neurons after stimulation with convulsant drugs. These findings further our understanding of seizure-induced nuclear events in neurons which are relevant to epileptogenesis. My study also demonstrates a robust nuclear profiling method amenable to different sequencing platforms for other cell types.