title: ClC-3 chloride channel in hippocampal neuronal apoptosis.
abstract: Over-production of nitric oxide is pathogenic for neuronal apoptosis around the ischemic area fol-lowing ischemic brain injury. In this study, an apoptotic model in rat hippocampal neurons was tablished by 0.5 mmol/L 3-morpholinosyndnomine (SIN-1), a nitric oxide donor. The models were then cultured with 0.1 mmol/L of 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS; the chloride channel blocker) for 18 hours. Neuronal survival was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and apoptosis was assayed by Hoechst 33342-labeled neuronal DNA fluorescence staining. Western blot analysis and immunoche-nescence staining were applied to determine the changes of activated caspase-3 and CIC-3 channel proteins. Real-time PCR was used to detect the mRNA expression of CIC-3. The results showed that SIN-1 reduced the neuronal survival rate, induced neuronal apoptosis, and promoted ClC-3 chloride channel protein and mRNA expression in the apoptotic neurons. DIDS reversed the effect of SIN-1. Our findings indicate that the increased activities of the ClC-3 chloride channel may be involved in hippocampal neuronal apoptosis induced by nitric oxide.
authors.affiliation: Department of Physiology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China ; First Hospital of Putian City, Putian 351100, Fujian Province, China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Pediatrics, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
authors.forename: Shuling
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Department of Physiology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
authors.forename: Hongling
authors.initials: H
authors.lastname: Fan
authors.affiliation: Department of Physiology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
authors.forename: Zhichao
authors.initials: Z
authors.lastname: Zhong
authors.affiliation: Department of Physiology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Physiology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: Jin
authors.affiliation: Department of Physiology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
authors.forename: Quanzhong
authors.initials: Q
authors.lastname: Chang
keywords: 3-morpholinosyndnomine; 4,4’-diisothiocyanostilbene-2,2’-disulfonic acid; ClC-3 chloride channel; apoptosis; brain injury; grants-supported paper; hippocampal neurons; neural regeneration; neuroregeneration; nitric oxide