Ontogenic changes and differential localization of T-type Ca2+ channel subunits Cav3.1 and Cav3.2 in mouse hippocampus and cerebellum

• Main Authors: Carolina Aguado, Sebastián García-Madrona, Mercedes Gil-Minguez, Rafael Lujan
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2016-08-01
• Series: Frontiers in Neuroanatomy
• Subjects: Calcium Channels; Cerebellum; Hippocampus; Immunohistochemistry; Electron microscopy; quantification
• Online Access: http://journal.frontiersin.org/Journal/10.3389/fnana.2016.00083/full

T-type calcium channels play a central role in regulating membrane excitability in the brain. Although the contributions of T-type current to neuron output is often proposed to reflect a differential distribution of T-type channel subtypes to somato-dendritic compartments, their precise subcellular distributions in central neurons are not fully determined. Using histoblot and high-resolution immunoelectron microscopic techniques, we have investigated the expression, regional distribution and subcellular localization of Cav3.1 and Cav3.2 subunits in the adult brain, as well as the ontogeny of expression during postnatal development. Histoblot analysis showed that Cav3.1 and Cav3.2 proteins were widely expressed in the brain, with mostly non-overlapping patterns. Cav3.1 showed the highest expression level in the molecular layer of the cerebellum, and Cav3.2 in the hippocampus and the molecular layer of cerebellum. During development, levels of Cav3.1 and Cav3.2 increased with age, although there were marked region- and developmental stage-specific differences in their expression. At the cellular and subcellular level, immunoelectron microscopy showed that labelling for Cav3.1 was present in somato-dendritic domains of hippocampal interneurons and Purkinje cells, while Cav3.2 was present in somato-dendritic domains of CA1 pyramidal cells, hippocampal interneurons and Purkinje cells. Most of the immunoparticles for Cav3.1 and Cav3.2 were either associated with the plasma membrane or the intracellular membranes, with notable differences depending on the compartment. Thus, Cav3.1 was mainly located in the plasma membrane of interneurons, whereas Cav3.2 was mainly located in the plasma membrane of dendritic spines and had a major intracellular distribution in dendritic shafts. In Purkinje cells, Cav3.1 and Cav3.2 showed similar distribution patterns. In addition to its main postsynaptic distribution, Cav3.2 but not Cav3.1 was also detected in axon terminals establishing excitatory synapses. These results shed new light on the subcellular localization of T-type channel subunits and provide evidence for the non-uniform distribution of Cav3.1 and Cav3.2 subunits over the plasma membrane of central neurons, which may account for the functional heterogeneity of T-type mediated current.