Subcellular distribution of low-voltage activated T-type Ca2+ channel subunits (Ca(v)3.1 and Ca(v)3.3) in reticular thalamic neurons of the cat.
Low-voltage-activated (LVA) Ca(2+) channels play a critical role in the generation of burst firing in the thalamus. Recently, three LVA Ca(2+) channel isoforms (Ca(v)3.1, Ca(v)3.2, Ca(v)3.3) have been identified in the reticular thalamic nucleus (RE). Previous electrophysiological and modelling studies have suggested that kinetically different T-type channels might be expressed in a compartmentalized manner in RE cells. However, their precise subcellular distribution has not been fully elucidated. Using light and electron microscopic (EM) immunocytochemistry, we investigated the subcellular expression pattern of Ca(v)3.1 and Ca(v)3.3 channel subunits in RE neurons of the cat. Fluorescent and peroxidase labelling demonstrated the presence of Ca(v)3.1 channel predominantly on the somata and proximal dendrites and Ca(v)3.3 channels on cell bodies. Quantitative immunogold localization disclosed that Ca(v)3.1 and Ca(v)3.3 isoforms showed 5.8- and 8.7-fold higher density, respectively, in the cytoplasm compared with somatic plasma membrane. Density of Ca(v)3.1 isoform in the somatic plasma membrane was 2.21-fold higher compared with Ca(v)3.3 subunit. In the dendritic plasma membrane, Ca(v)3.1 channel isoform was expressed throughout the entire dendritic tree. In contrast, Ca(v)3.3 isoform was absent from large-caliber, presumably proximal dendritic segments. Quantitative comparison showed that the relative density of immunogold particles compared with dendritic surface was 8.9- and 14.8-fold higher for Ca(v)3.1 and Ca(v)3.3, respectively, in small-diameter dendrites than in large proximal dendritic segments or somata. Our results demonstrate a higher density of low-threshold Ca(2+) channels in distal dendrites and provide further evidence of the role of RE neuron dendrites in the generation of prolonged, low-threshold spike bursts.
Centre de Recherche Université Laval Robert-Giffard, Québec, Québec, Canada.
Krisztina
K
Kovács
Attila
A
Sík
Christopher
C
Ricketts
Igor
I
Timofeev
D000818:Animals; D015153:Blotting, Western; D020747:Calcium Channels, T-Type / Q000378:metabolism*; D002415:Cats; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003593:Cytoplasm / Q000378:metabolism / Q000648:ultrastructure; D003712:Dendrites / Q000378:metabolism / Q000648:ultrastructure; D005453:Fluorescence; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D020646:Intralaminar Thalamic Nuclei / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D010320:Parvalbumins / Q000378:metabolism; D020033:Protein Isoforms / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure; D013787:Thalamic Nuclei / Q000378:metabolism / Q000648:ultrastructure
D020747:Calcium Channels, T-Type; D010320:Parvalbumins; D020033:Protein Isoforms