Immunocytochemical localization of nicotinic acetylcholine receptor in the rat cerebellar cortex

Although nicotinic acetylcholine receptors (nAChRs) have been reported to function in the cerebellar cortex, nicotinic acetylcholine receptor subtypes contributing to nicotinic currents in the cortex have not been well characterized at the subcellular level. This study deals with immunocytochemical localization of the nicotinic acetylcholine receptor alpha4 subunit using a monoclonal antibody against the alpha4 subunit. Alpha4-LI (alpha4-like immunoreactivity) was detected in the cell bodies of molecular, Purkinje cell and granular layers. In particular, the cell bodies of Purkinje cells were extensively immunostained. In Purkinje cells, alpha4-LI was found in perikarya mainly associated with rough endoplasmic reticulum, plasma membrane, and cytoplasmic matrix. At higher magnification, the immunoreaction product was densely localized along with somatic plasma membranes at the axo-somatic synapse and the plasma membranes at extrasynaptic regions of cell bodies. Alpha4-LI was also found in the axon terminals which form synapses with Purkinje cells. In the granular layer, somatic cell membranes of granular cells were immunostained. These morphological observations suggest that alpha4-containing nAChRs contribute nicotinic currents reported in Purkinje cells, and that presynaptic alpha4-containing nAChRs regulate the release of neurotransmitters on the axon terminals found near Purkinje cells.