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Rationale: Developed an antibody against the pore forming subunit of Cav1.2 that was used in light and electron microscopy of fixed and stained rat brain sections. Fig. 1 shows characterization and validation of antibody specificity. Immunoreactivity against the Cav1.2 pore-forming subunit (Cav1.2-IR) was observed in both myelinated and unmyelinated axons. Within labeled axons, Cav1.2-IR was either clustered or filled the intracellular space (Fig. 5A). Axon terminals were also labeled, especially in CA1 stratum radiatum (Fig. 5B) and dentate gyrus (DG) stratum moleculare and least in CA3 stratum lucidum (Table 1). A majority of the labeled axon terminals formed asymmetric synapses. Cav1.2-IR was also observed in a large fraction of mossy fiber terminals in CA3 stratum lucidum and DG central hilus (Table 1), which generally formed asymmetric contacts with unlabeled dendrites (Fig. 5C,D). In these terminals, Cav1.2-IR formed clusters along the plasma membrane at synaptic and nonsynaptic sites but was also detected near nonsynaptic junctions with unlabeled dendrites resembling puncta adherentia. 21/6/2017 Pim - I regard the predominant intracellular localization of Cav21 in these preparation as reflecting a non-canonical localization for this protein (protein synthesis, inactive pool) and I refrain from annotation of this localization.. Experimental description: The brains of adult male Sprague-Dawley rats (2 months old) were fixed and cut into coronal sections with a vibrating microtome. Mutant mice with targeted inactivation of Cav1.2 expression in the hippocampus (Cav1.2HCKO) were used in antibody verification.
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