Synaptic localization of ionotropic glutamate receptors in the rat substantia nigra

• Published in: Neuroscience 2000-01, Vol.101 (4), p.1037-1051
• Main Authors: Chatha, B.T, Bernard, V, Streit, P, Bolam, J.P
• Format: Article
• Language: English
• Subjects: AMPA receptors; Animals; basal ganglia; Biological and medical sciences; Central nervous system; Central neurotransmission. Neuromudulation. Pathways and receptors; Fundamental and applied biological sciences. Psychology; Immunohistochemistry; immunolabelling; Microscopy, Electron; NMDA receptors; Protein Isoforms - metabolism; Rats; Rats, Wistar; Receptors, AMPA - metabolism; Receptors, Glutamate - metabolism; Receptors, N-Methyl-D-Aspartate - metabolism; Substantia Nigra - metabolism; Substantia Nigra - ultrastructure; synapses; Synapses - metabolism; Synapses - ultrastructure; Tissue Distribution; Vertebrates: nervous system and sense organs
• ISSN: 0306-4522; 1873-7544
• DOI: 10.1016/S0306-4522(00)00432-2

Glutamatergic neurotransmission in the substantia nigra pars compacta and pars reticulata is mediated through N-methyl- d-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxaline propionic acid/kainate (AMPA) type receptors as well as other glutamate receptors and is critical for basal ganglia functioning. A major glutamatergic input to the substantia nigra originates in the subthalamic nucleus, and the long-lasting stimulation of the dopaminergic cells of the substantia nigra pars compacta by the subthalamic neurons has been implicated in the pathophysiology of Parkinson’s disease. The objectives of the present study were to determine the subcellular and subsynaptic localization of subunits of the N-methyl- d-aspartate and AMPA receptors in the substantia nigra, and also to determine whether co-localization of N-methyl- d-aspartate and AMPA receptor subunits occur at individual synapses. To achieve this, pre-embedding and post-embedding immunocytochemistry was applied to sections of substantia nigra using antibodies that recognize the NR1 and NR2A/B subunits of the N-methyl- d-aspartate receptor, and GluR2/3 subunits of the AMPA receptor. In both regions of the substantia nigra, immunolabelling for each of the subunits was observed in numerous perikarya and proximal dendrites. At the subcellular level, silver-intensified immunogold particles localizing N-methyl- d-aspartate and AMPA receptor subunits were most commonly present within dendrites where they were associated with a variety of intracellular organelles and with the internal surface of the plasma membrane. Post-embedding immunogold labelling revealed immunoparticles labelling for NR1, NR2A/B and GluR2/3 to be enriched at asymmetric synaptic specializations, although a large proportion of asymmetric synapses were immunonegative. Double immunolabelling revealed, in addition to single-labelled synapses, the co-localization of subunits of the N-methyl- d-aspartate receptor and subunits of the AMPA receptor at individual asymmetric synapses. Similarly, double immunolabelling also revealed the co-localization of the NRl and NR2A/B subunits of the N-methyl- d-aspartate receptor at individual asymmetric synapses. Labelling for NR1 and GluR2/3 was, on average, relatively evenly distributed across the width of the synapse with a gradual reduction towards the periphery when analysed in single sections. In summary, the present results demonstrate that AMPA and N-methyl- d-aspartate receptors are selectively loca