title: Expression and cellular localization of HCN channels in rat cerebellar granule neurons.
abstract: 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.
authors: Centro de Investigaciones Médicas CIM, Escuela de Medicina, Universidad de Talca, Talca, Chile.
authors: Rafael
authors: R
authors: Zúñiga
authors: Departamento de Fisiología, Universidad de Concepción, Concepción, Chile.
authors: Daniela
authors: D
authors: González
authors: Centro de Investigaciones Médicas CIM, Escuela de Medicina, Universidad de Talca, Talca, Chile.
authors: Claudio
authors: C
authors: Valenzuela
authors: Centro de Investigaciones Médicas CIM, Escuela de Medicina, Universidad de Talca, Talca, Chile.
authors: Nelson
authors: N
authors: Brown
authors: Centro de Investigaciones Médicas CIM, Escuela de Medicina, Universidad de Talca, Talca, Chile. Electronic address: lzuniga@utalca.cl.
authors: Leandro
authors: L
authors: Zúñiga
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002531:Cerebellum / Q000378:metabolism*; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000235:genetics / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D009474:Neurons / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; D021122:Protein Subunits; D012333:RNA, Messenger
keywords: Cerebellar granule neurons (CGN); HCN channels; HCN channels in cerebellum; Hyperpolarization-activated cyclic nucleotide-gated channels