Acid sensing ion channel la involving in development of cerebella Purkinje cells

Objective To investigate the mechanism of acid sensing ion channel (ASIC) 1a in the development of Purkinje cells in the cerebellum. Methods Newborn C57BL/6 mice were chosen and Purkinje cells were obtained from these mice by in vitro culture. Purkinje cells were divided into experimental group and control group, and shRNA-ASIC1a or shRNA-control sequence was used to construct recombinant plasmid lentivirus infections at the early and late stages of cell developments. The morphological structures of Purkinje cells were detected by immunofluorescent staining and the dendritic branches were counted at the early and late stages of cell developments. Western blotting was employed to detect the calcium binding protein D-28K, glial fibrillary acidic protein, Zic, parvalbumin and N-methyl-D-aspartate receptor (NMDAR) expressions. Real time(RT)-PCR was used to detect the expressions of endoplasmic reticulum stress related factor CCAAT/ enhancer binding protein homologous protein CHOP and protein kinase R- like ER kinase (PERK). Results Immunofluorescence indicated that Purkinje cells showed obvious morphological differences between the experimental group and control group; the dendrite growth and development in the control group were significantly better than those in the experimental group (P<0.05); the number of 2nd and 3rd stage dendritic branches of Purkinje cells in the control group was significantly larger than that in the experimental group (P<0.05). Western blotting showed that D- 28K expression in the Purkinje cells of experimental group was significantly decreased as compared with that in the control group at the early stage (P<0.05); NMDAR expression in the Purkinje cells of experimental group was significantly increased as compared with that in the control group at the early and late stages (P<0.05). RT-PCR results showed that CHOP and PERK expressions in the Purkinje cells of experimental group were significantly higher than those in the control group (P< 0.05). Conclusion ASIC1a has an important role in the development of cerebella Purkinje cells. Key words: Purkinje cell; Acid sensing ion channel 1a; Endoplasmic reticulum stress; N-methyl-D-aspartate receptor