Expression and cellular localization of HCN channels in rat cerebellar granule neurons

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• 作者：Rafael Zú ; ñ ; iga^a ; Daniela Gonzá ; lez^b ; Claudio Valenzuela^a ; Nelson Brown^a ; Leandro Zú ; ñ ; iga^a ; ^{lzuniga@utalca.cl" class="auth_mail" title="E-mail the corresponding author}
• 关键词：HCN channels ; Hyperpolarization-activated cyclic nucleotide-gated channels ; Cerebellar granule neurons (CGN) ; HCN channels in cerebellum
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2016
• 出版时间：23 September 2016
• 年：2016
• 卷：478
• 期：3
• 页码：1429-1435
• 全文大小：1914 K

文摘

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels belong to the superfamily of voltage-gated pore loop channels. In mammals, this family consists of four different subunits (HCN1–4) and their ion channels activity have been proposed to play an essential role in regulating the membrane potential of excitable cells. Here, we describe the expression and relative abundances of HCN channels in cerebellum and primary cultures of cerebellar granule neurons (CGN). Quantitative determination of mRNA expression levels demonstrated the existence of an accumulation pattern of transcripts in cerebellum that encode HCN2 > HCN3 = HCN4 > HCN1 subunits. Immunolocalization analyses of HCN channels in cerebella revealed positive staining in Purkinje and granule cell layers. The presence of the HCN subunits in the cerebellar granule cell layer was then confirmed in primary cultures of CGN by quantitative real-time PCR (qPCR), as well as western blot and immunofluorescence analysis, demonstrating the presence of all four channel proteins.