Localization of AMPA-selective glutamate receptor subunits in the adult cat visual cortex.

We have studied the presence and distribution of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-selective glutamate receptor subunits (GluR1, 2, 3, and 4) in the adult cat visual cortical areas 17, 18, 19, and the lateral suprasylvian areas (LSA). Reverse transcription-polymerase chain reaction (RT-PCR) amplification indicated that the genes encoding GluR1, 2, 3, and 4 are expressed in these areas and Western blot analysis revealed that the size of the corresponding peptides is similar to those described in the rat brain. In situ hybridization (ISH) using digoxigenin-labeled riboprobes showed that mRNAs coding for GluR1 and GluR3 were located in cells in all layers of the areas examined and also in the underlying white matter. GluR1 mRNA was relatively abundant throughout layers II-VI while GluR3 mRNA revealed a more laminated pattern of expression, preferentially labeling cells in layers II, III, V, and VI. The distribution of AMPA-selective receptor subunit peptides was studied by immunohistochemistry using subunit specific antibodies and found to be consistent with ISH results. In addition, we observed that most of the cells strongly labeled by the anti-GluR1 antibody were non-pyramidal neurons and that intense GluR2/3 immunoreactivity was seen preferentially in pyramidal neurons. Interestingly, double-labeling experiments indicated that neurons expressing gamma-aminobutyric acid (GABA) as well as the GluR1 subunit were particularly abundant in deeper layers. The GluR4 peptide was predominantly found in a relatively low number of layer III and layer V neurons with either pyramidal or non-pyramidal morphology. Finally, the distribution of neurons expressing the various receptor subunits was similar in all the visual cortical areas studied. These findings indicate a high expression of GluR1-3 subunits in the cat visual cortex and that GluR1 and GluR2/3 subunits are particularly abundant in non-pyramidal and pyramidal neurons, respectively. In addition, the results described here provide a reference for future studies dealing with the effect of visual deprivation on the expression of this receptor type.

Glutamate receptors in astrocytic end-feet.

Glial cells in the mammalian central nervous system, like neurons, possess glutamate receptors suggesting that neuronal-glial communication via neurotransmitters is an important component of brain function. In the bovine corpus callosum, GLUR-1, an alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA)/kainate receptor subtype, is a major type of glutamate receptor. It is distributed along astrocytic processes and it is particularly abundant in the end-feet and the glial fibers surrounding the capillaries. This close spatial relationship between GLUR-1 receptors in astrocytes and endothelial cells suggests that these receptors may be activated by glutamate transported across the blood-brain barrier and thus regulate the barrier itself by ensuing astrocyte-endothelial cell interactions.

Distribution of neurons expressing substance P receptor messenger RNA in immature and adult cat visual cortex.

We have investigated the expression of messenger RNA (mRNA) encoding substance P receptor (SPR) in the visual cortex of adult cats and 17-day-old kittens, using in situ hybridization histochemistry with two digoxigenin-labeled oligodeoxynucleotides complementary to the SPR mRNA. In the adult cortex, a subset of large pyramidal neurons of layer V and layer III is heavily labeled. Other, mainly pyramidal neurons in layers II, III and V are less intensely labeled, but most neurons in these layers appear unlabeled. Neurons in layer IV and VI, and in the white matter do not show hybridization signals above background levels. In the 17-day-old kitten, SPR mRNA-expressing cells are confined to layer V and to the upper white matter (subplate zone), whereas supragranular neurons do not yet contain SPR mRNA. A few neurons in layer VI display moderate labeling. Astrocytes, identified with anti-glial fibrillary acid protein antibodies, did not express detectable levels of SPR mRNA in both adult and kitten visual cortex. These results indicate that SPR mRNA expression is transient in neurons of the white matter, and is developmentally regulated in supragranular layers. In addition, the localization of SPR mRNA in a subset of pyramidal cells suggests that substance P modulates the excitability of certain projection neurons which are the origin of extrinsic connections.