Sparse colocalization of somatostatin- and GABA-immunoreactivity in the entorhinal cortex of the rat

We studied the regional and laminar distribution of neurons expressing immunoreactivity with antibodies against the neuropeptide somatostatin (SOM) in the entorhinal cortex of colchicine-treated rats. We further determined whether these neurons also express immunoreactivity with antibodies against the neurotransmitter gamma-aminobutyric acid (GABA). Frontally and horizontally cut brain sections were subjected to double immunofluorescence histochemistry and investigated in a two-laser confocal laser scanning fluorescence microscope. The exact position of each single- or double-labeled cell was obtained via the preparation of large-scale digital fluorescence images superimposed on a brightfield digital image obtained postscanning after decoverslipping and staining with cresyl violet. Three types of SOM-positive cells were found: big multipolar cells (10-15% of the SOM-positive cells), oval cells (15-20%), and small spherical cells (majority of SOM-positive cells). Most cells were seen in layer III. In addition, we found immunoreactive cells in the other layers, with the fewest cells in layers I and IV (lamina dissecans). Of the SOM-positive cells, 18% also expressed GABA immunoreactivity; of the GABA-positive cells, 8% were also immunoreactive for SOM. Double-labeled cells were mostly small spherical cells and, infrequently, multipolar. These data indicate that in the entorhinal cortex, a large proportion of the cells belonging to the SOM population do not express GABA. We speculate that there may be several subpopulations of SOM cells, of which the largest may consist of non-GABAergic, excitatory interneurons.