Expression and cellular localization of HCN channels in rat cerebellar granule neurons.
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.
['Zúñiga|Rafael|R|', 'González|Daniela|D|', 'Valenzuela|Claudio|C|', 'Brown|Nelson|N|', 'Zúñiga|Leandro|L|']
D000818:Animals D015153:Blotting, Western D002531:Cerebellum D054815:Cyclic Nucleotide-Gated Cation Channels D003594:Cytoplasmic Granules D005455:Fluorescent Antibody Technique D007150:Immunohistochemistry D009474:Neurons D021122:Protein Subunits D021381:Protein Transport D012333:RNA, Messenger D017207:Rats, Sprague-Dawley D060888:Real-Time Polymerase Chain Reaction
HCN channels HCN channels in cerebellum Cerebellar granule neurons (CGN) Hyperpolarization-activated cyclic nucleotide-gated channels