title: Exploration of the Brn4-regulated genes enhancing adult hippocampal neurogenesis by RNA sequencing.
abstract: Adult hippocampal neurogenesis is essential for learning and memory, and its dysfunction is involved in neurodegenerative diseases. However, the molecular mechanisms underlying adult hippocampal neurogenesis are still largely unknown. Our previous studies indicated that the transcription factor Brn4 was upregulated and promoted neuronal differentiation of neural stem cells (NSCs) in the surgically denervated hippocampus in rats. In this study, we use high-throughput RNA sequencing to explore the molecular mechanisms underlying the enhancement of adult hippocampal neurogenesis induced by lentivirus-mediated Brn4 overexpression in vivo. After 10 days of the lentivirus injection, we found that the expression levels of genes related to neuronal development and maturation were significantly increased and the expression levels of genes related to NSC maintenance were significantly decreased, indicating enhanced neurogenesis in the hippocampus after Brn4 overexpression. Through RNA sequencing, we found that 658 genes were differentially expressed in the Brn4-overexpressed hippocampi compared with GFP-overexpressed controls. Many of these differentially expressed genes are involved in NSC division and differentiation. By using quantitative real-time PCR, we validated the expression changes of three genes, including Ctbp2, Notch2, and Gli1, all of which are reported to play key roles in neuronal differentiation of NSCs. Importantly, the expression levels of Ctbp2 and Notch2 were also significantly changed in the hippocampus of Brn4 KO mice, which indicates that the expression levels of Ctbp2 and Notch2 may be directly regulated by Brn4. Our current study provides a solid foundation for further investigation and identifies Ctbp2 and Notch2 as possible downstream targets of Brn4. © 2017 Wiley Periodicals, Inc.
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Guo
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Cheng
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Linmei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Mao
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Guixiang
authors.initials: G
authors.lastname: Tian
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Wenhao
authors.initials: W
authors.lastname: Xu
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Yuhao
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Ma
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Jianbing
authors.initials: J
authors.lastname: Qin
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.
authors.forename: Meiling
authors.initials: M
authors.lastname: Tian
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.|Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu, China.
authors.forename: Guohua
authors.initials: G
authors.lastname: Jin
authors.affiliation: Neurosurgery Department, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Shi
authors.affiliation: Department of Anatomy, Nantong University, Nantong, Jiangsu, China.|Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu, China.
authors.forename: Xinhua
authors.identifier: 0000-0002-5702-6733
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D005786:Gene Expression Regulation / Q000502:physiology*; D059014:High-Throughput Nucleotide Sequencing; D006624:Hippocampus; D051379:Mice; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D055495:Neurogenesis / Q000502:physiology*; D050809:POU Domain Factors / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D009419:Nerve Tissue Proteins; D050809:POU Domain Factors; C484088:Pou3f4 protein, rat
keywords: Brn4; RNA sequencing; hippocampus; neurogenesis