A monoclonal antibody to conotoxin reveals the distribution of a subset of calcium channels in the rat cerebellar cortex

Abstract

Voltage-sensitive calcium channels (VSCC) are a family of ionophores having different electrical and pharmacological properties. The ω-conotoxin GVIA (ω-CgTX) is a specific blocker of one subset of VSCCs. Because of the specificity of this toxin, a monoclonal anti-ω-CgTX antibody was generated against a ω-CgTX-key hole limpet hemocyanin conjugate and used as a specific marker to study VSCC distributions. This mab was shown to recognize ω-CgTX on Western blots and to display ω-CgTX-dependent immunoperoxidase staining of rat cerebellum. Incubation of fresh, unfixed sections of adult rat cerebellum in ω-CgTX followed by light fixation and peroxidase immunocytochemistry with mab anti-ω-CgTX revealed a specific pattern of labelling. All principal classes of cerebellar neurons were immunoreactive, but in general glial cells were not stained. Most interestingly, strong focal immunoreactivity was encountered at branching points of Purkinje cell dendrites. This characteristic staining pattern implies that a subset of VSCC is specifically concentrated in these regions and suggests that these channels may play a role in the functional integration of dendritic signals.