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Non-NMDA excitatory amino acid receptors in a subcellular fraction enriched in cerebellar glomeruli

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Abstract

In the internal granular layer of the cerebellar cortex the polysynaptic complexes called glomeruli consist mainly of homogeneous populations of glutamatergic and GABAergic synapses, both located on granule cell dendrites. A subcellular fraction enriched in glomeruli was prepared from rat cerebellum, and the distribution of the different types of NMDA and non-NMDA glutamate binding sites was studied in the membranes derived from this fraction (fraction G) as compared to that in the membranes prepared from a total cerebellar homogenate (fraction T). Cl−/Ca2+ independent [3H]glutamate binding sites were not abundant and could be reliably measured only in fraction G. Cl− dependent/Ca2+ activated [3H]glutamate binding sites were more abundant and exhibited a single K d in both fractions G and T. Quisqualate, NMDA, kainate, L-AP4 andtrans-ACPD inhibited [3H]glutamate binding to different extents in the two membrane fractions. Quisqualate sensitive sites were predominant in all cases but more abundant in fraction T than in fraction G. An opposite distribution was observed for the NMDA sensitive binding sites while kainate sensitive binding sites were scarce everywhere.Trans-ACPD, a ligand presumed selective for metabotropic glutamate binding sites, displaced [3H]glutamate from fraction T but nor from fraction G, suggesting the absence of these sites from glomeruli. Similarly, no L-AP4 sensitive sites were present in fraction G while they were abundant in fraction T. Binding sites associated with ionotropic receptors of the quisqualate type were determined by measuring [3H]AMPA binding. The density of the high affinity [3H]AMPA binding sites in fraction T was twice as high as in fraction G, indicating that these sites are abundant in structures other than glomeruli. High-affinity [3H]kainate binding sites are more abundant in fraction G than in fraction T; the same, but with smaller differences, occurs for the distribution of the low affinity [3H]kainate binding sites. The density of the latter sites is close to that of the high affinity [3H]AMPA binding sites confirming the presence of quisqualate/kainate receptors on granule cells, as previously hypothesized (for review, see Gallo et al., 1990). Taken together, these results indicate a segregation of the glutamate binding sites types at specialized synapses or neuronal cell types in the cerebellar network.

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Abbreviations

AMPA:

(RS)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid

DL-AP4:

dl-2-amino-4-phosphonobutyric acid

D-AP5:

d-2-amino-5-phosphonovaleric acid

EAA:

excitatory amino acid

EGTA:

ethylene glycol-bis(β-aminoethyle ether) N,N,N′,N′-tetracetic acid

NMDA:

N-methyl-D-aspartate

Quisqualate:

β-[3,5-dioxo-1,2,4-oxadiazolidin-2-yl]-L-alanine

trans-ACPD:

trans-1-amino-cyclopentyl-1,3-dicarboxylic acid

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Authors and Affiliations

  1. Centre de Neurochimie du CNRS, 5 Rue Blaise Pascal, F-67000, Strausbourg, France

    F. Viennot, J. de Barry & G. Gombos

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  1. F. Viennot
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  2. J. de Barry
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  3. G. Gombos
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Viennot, F., de Barry, J. & Gombos, G. Non-NMDA excitatory amino acid receptors in a subcellular fraction enriched in cerebellar glomeruli. Neurochem Res 16, 435–442 (1991). https://doi.org/10.1007/BF00965563

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