Immunohistochemical localization of N-methyl- d-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunits in the substantia nigra pars compacta of the rat

Abstract

Ionotropic glutamate receptors in the substantia nigra pars compacta regulate the activity of dopamine neurons. We have used dual-label immunofluoresence and confocal laser microscopy to study the localization of subunits of two types of ionotropic receptors within the substantia nigra pars compacta of the rat. Immunostaining for N-methyl- d-aspartate receptor 1 and glutamate receptor2/3 was prominent in the soma and proximal dendrites of all tyrosine hydroxylase-immunopositive cells, while only low amounts of N-methyl- d-aspartate receptor 2A and N-methyl- d-aspartate receptor 2B were present. Selective antibodies were used to determine the isoforms of N-methyl- d-aspartate receptor 1 present. Immunostaining for the NI, CI and C2 variably spliced segments of N-methyl- d-aspartate receptor 1 were scarce in the substantia nigra pars compacta, while immunoreactivity for the alternative C2′ terminus of N-methyl- d-aspartate receptor 1 was quite abundant. Staining for glutamate receptor 1 was heterogeneous; about half of the tyrosine hydroxylase immunopositive cells stained intensely, while the other half were immunonegative. The glutamate receptor 1-stained cells were concentrated in the ventral tier of the substantia nigra pars compacta. Glutamate receptor 4 was not found in tyrosine hydroxylase-immunopositive cells within the substantia nigra pars compacta. Together, these data demonstrate that dopaminergic neurons in the substantia nigra pars compacta express primarily glutamate receptor 1, glutamate receptor2/3 and N-methyl- d-aspartate receptor 1 isoforms containing the alternative C2′ terminus.