title: Small RNA interference-mediated gene silencing of TREK-1 potassium channel in cultured astrocytes.
abstract: This study was aimed to examine the effect of TREK-1 silencing on the function of astrocytes. Three 21-nucleotide small interfering RNA (siRNA) duplexes (siT1, siT2, siT3) targeting TREK-1 were constructed. Cy3-labeled dsRNA oligmers were used to determine the transfection efficiency in cultured astrocytes. TREK-1-specific siRNA duplexes (siT1, siT2, siT3) at the optimal concentration were transfected into cultured astrocytes, and the most efficient siRNA was identified by the method of immunocytochemical staining and Western blotting. The proliferation of astrocytes tranfected with TREK-1-targeting siRNA under hypoxia condition was measured by fluorescence-activated cell sorting (FACS). The results showed that TREK-1 was expressed in cultured astrocytes. The dsRNA oligmers targeting TREK-1 could be transfected efficiently in cultured astrocytes and down-regulate the expression of TREK-1 in astrocytes. Moreover, the down-regulation of TREK-1 in astrocytes contributed to the proliferation of astrocytes under hypoxia condition as determined by cell cycle analysis. It was concluded that siRNA is a powerful technique that can be used to knockdown the expression of TREK-1 in astrocytes, which helps further investigate the function of TREK-1 channel in astrocytes under physicological and pathological condition.
authors.affiliation: Department of Neurology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Neurology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Ronghua
authors.initials: R
authors.lastname: Tang
authors.affiliation: Department of Neurology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Division of Clinical Immunology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Jingjiao
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Neurology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Department of Neurology, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Neurology, Huazhong University of Science and Technology, Wuhan, 430030, China. xiemj2000@yahoo.com.cn.
authors.forename: Minjie
authors.initials: M
authors.lastname: Xie
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000502:physiology*; D002478:Cells, Cultured; D020868:Gene Silencing / Q000502:physiology*; D015221:Potassium Channels; D024683:Potassium Channels, Tandem Pore Domain / Q000235:genetics*; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats
chemical_list: D015221:Potassium Channels; D024683:Potassium Channels, Tandem Pore Domain; D034741:RNA, Small Interfering; C103742:potassium channel protein TREK-1