Calbindin D 28k-containing nonpyramidal cells in the rat hippocampus: Their immunoreactivity for GABA and projection to the medial septum

Abstract

Calbindin D 28k-containing non-pyramidal cells were found in all layers and subfields of the hippocampus, with the highest frequency in stratum radiatum of the CA1–CA3 subfields. A large number of these neurons had a vertically oriented dendritic tree, often restricted to stratum radiatum. In stratum oriens and near to the border of strata radiatum and lacunosum moleculare cells with horizontally running dendrites were also found. Multipolar cells were most common in stratum radiatum of the CA3 region. The GABAergic nature of the calbindin D 28k-containing non-pyramidal cells was studied using the “mirror” technique. Adjacent thick sections were immunostained for calbindin D 28k and GABA, and halved neurons were identified on the common surfaces. The majority of calbindin D 28k-containing non-pyramidal cells were shown to be GABAergic. The GABA-negative calbindin cells were found in relatively large numbers in stratum oriens of the CA1–CA3 region, and occasionally in strata radiatum and pyramidale of CA2, and in stratum radiatum of the CA3c region near to the border of the dentate hilus. However, even in these cells a weak immunostaining, only slightly but consistently above background level, was always observed. Earlier studies have demonstrated that the somata of GABAergic neurons with distant projections may contain a level of GABA that is below the detection threshold of immunocytochemistry. Here we provide direct evidence that the calbindin-containing non-pyramidal cells were among those projecting to the medial septum. Following horseradish peroxidase injections into the medial septum 80% of the retrogradely labelled non-pyramidal cells were found to be immunoreactive for calbindin D 28k, and 20% contained neuropeptide Y. These results suggest that the calbindin D 28k-containing and apparently GABA-immunonegative non-pyramidal cells in stratum oriens of the CAl–CA3 regions may also be GABAergic, but have a distant projection, that is, to the medial septum.