title: Nr3C1-Bhlhb2 Axis Dysregulation Is Involved in the Development of Attention Deficit Hyperactivity.
abstract: Attention deficit hyperactivity disorder (ADHD) is a child developmental and behavioral disorder which seriously hinders their education and development. To investigate the key regulators in the prefrontal cortex (PFC), the major affected areas of ADHD, microRNA (miR)-138,138*, 34c*, 296, and 494, were noted for their significant downregulation in ADHD model rats spontaneously hypertensive rats (SHRs) compared to Wistar Kyoto (WKY) rat control. Based on promoter sequence analysis and activity assay, glucocorticoid receptor (Nr3c1) was identified for the inhibition of the promoter activity of miR-138-1, 34c*, 296, and 494 genes and their transcription. In the PFC of ADHD model rats SHR, Nr3c1 expression was abnormally elevated and reversely correlated with the levels of miR-138-1, 34c, 296, and 494 expression. Luciferase report assays indicated that all miR-138, 138*, 34c*, 296, and 494 targeted the 3' untranslated region of transcription factor Bhlhb2 (Bhlhe40) messenger RNA (mRNA) in common and ectopic expression of miR-138,138*, 34c*, 296, and 494 further suppressed the expression of Bhlhb2 gene. Consistently, Bhlhb2 expression was significantly higher in PFC of ADHD model SHR than control. Overexpressed Bhlhb2 in vitro significantly suppressed PC12 cell differentiation, and silence of Bhlhb2 enhanced the growth of neurite axon and dendrite. To observe the roles of Bhlhb2 further in vivo, Bhlhb2 was silenced in the PFC of nine SHR rats. Interestingly, knockdown of Bhlhb2 significantly improved the hyperactivity behaviors in SHRs compared to control. These findings show that Nr3c1-Bhlhb2 axis dysregulation was involved in the development of attention deficit and hyperactivity.
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China. jaemny@163.com.|Department of Clinic Medicine, Zhejiang Medical College, 481 Binwen Road, Binjiang College Zone, Hangzhou, 310053, China. jaemny@163.com.
authors.forename: Li Hui
authors.initials: LH
authors.lastname: Wu
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China.|Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Cheng
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China.|Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Yu
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China.|Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Bao Mei
authors.initials: BM
authors.lastname: He
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China.|Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China.|Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Yi Wei
authors.initials: YW
authors.lastname: Dong
authors.affiliation: Department of Children's Health Care, The Second Affiliated Hospital & Yu Ying Children's Hospital, Wenzhou Medical University, Wenzhou, China.
authors.forename: Xiao Ting
authors.initials: XT
authors.lastname: Shao
authors.affiliation: Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China.
authors.forename: Qian Qian
authors.initials: QQ
authors.lastname: Cai
authors.affiliation: Department of Neonatology, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, China.
authors.forename: Min
authors.initials: M
authors.lastname: Peng
authors.affiliation: Key Lab of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, China. xz_wu@shmu.edu.cn.
authors.forename: Xing Zhong
authors.initials: XZ
authors.lastname: Wu
mesh_terms: D000818:Animals; D001289:Attention Deficit Disorder with Hyperactivity / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology*; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000235:genetics*; D053263:Gene Regulatory Networks / Q000502:physiology; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008297:Male; D016716:PC12 Cells; D017397:Prefrontal Cortex / Q000378:metabolism / Q000503:physiopathology; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D011965:Receptors, Glucocorticoid / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D018398:Homeodomain Proteins; D011965:Receptors, Glucocorticoid; C491538:Bhlhe40 protein, rat; C542344:NR3C1 protein, rat
keywords: ADHD; DEC1; Glucocorticoid receptor; miRNA
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