title: Quantitative localization of Cav2.1 (P/Q-type) voltage-dependent calcium channels in Purkinje cells: somatodendritic gradient and distinct somatic coclustering with calcium-activated potassium channels.
abstract: P/Q-type voltage-dependent calcium channels play key roles in transmitter release, integration of dendritic signals, generation of dendritic spikes, and gene expression. High intracellular calcium concentration transient produced by these channels is restricted to tens to hundreds of nanometers from the channels. Therefore, precise localization of these channels along the plasma membrane was long sought to decipher how each neuronal cell function is controlled. Here, we analyzed the distribution of Ca(v)2.1 subunit of the P/Q-type channel using highly sensitive SDS-digested freeze-fracture replica labeling in the rat cerebellar Purkinje cells. The labeling efficiency was such that the number of immunogold particles in each parallel fiber active zone was comparable to that of functional channels calculated from previous reports. Two distinct patterns of Ca(v)2.1 distribution, scattered and clustered, were found in Purkinje cells. The scattered Ca(v)2.1 had a somatodendritic gradient with the density of immunogold particles increasing 2.5-fold from soma to distal dendrites. The other population with 74-fold higher density than the scattered particles was found within clusters of intramembrane particles on the P-face of soma and primary dendrites. Both populations of Ca(v)2.1 were found as early as P3 and increased in the second postnatal week to a mature level. Using double immunogold labeling, we found that virtually all of the Ca(v)2.1 clusters were colocalized with two types of calcium-activated potassium channels, BK and SK2, with the nearest neighbor distance of ∼40 nm. Calcium nanodomain created by the opening of Ca(v)2.1 channels likely activates the two channels that limit the extent of depolarization.
authors: Department of Physiological Sciences, Graduate University for Advanced Studies-Sokendai, Myodaiji, Okazaki 444-8787, Japan.
authors: Dwi Wahyu
authors: DW
authors: Indriati
authors: Naomi
authors: N
authors: Kamasawa
authors: Ko
authors: K
authors: Matsui
authors: Andrea L
authors: AL
authors: Meredith
authors: Masahiko
authors: M
authors: Watanabe
authors: Ryuichi
authors: R
authors: Shigemoto
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D020864:Calcium Channels, N-Type / Q000378:metabolism* / Q000648:ultrastructure; D003593:Cytoplasm / Q000737:chemistry / Q000648:ultrastructure; D003712:Dendrites / Q000378:metabolism* / Q000648:ultrastructure; D005614:Freeze Fracturing / Q000379:methods; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D018345:Mice, Knockout; D017729:Presynaptic Terminals / Q000378:metabolism / Q000648:ultrastructure; D011689:Purkinje Cells / Q000737:chemistry* / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D051640:Small-Conductance Calcium-Activated Potassium Channels / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D020864:Calcium Channels, N-Type; D051036:Large-Conductance Calcium-Activated Potassium Channels; D051640:Small-Conductance Calcium-Activated Potassium Channels; C496898:Kcnn2 protein, mouse