Comparative analysis of the excitatory and inhibitory hippocampal neurons activity during associative context memory retrieval

Abstract

In the present study, we analyzed the differential involvement of hippocampal interneurons and pyramidal neurons in the retrieval of associative aversive context memory. For this purpose, we used a model of associative learning in which the formation of a neutral context memory and the subsequent association of this memory with the footshock US during a brief reminder of the context were significantly separated in time. The activation of hippocampal neurons during associative context memory retrieval in this task was addressed by immunohistochemical detection of the immediate early gene c-fos protein. Retrieval of associative context memory was accompanied by an increase in the number of c-Fos-positive cells in the CA1 region, but not in the CA3 region and the dentate gyrus of the hippocampus. Next, a protein marker, the product of the homeobox-containing gene Emx1, was used to specifically identify excitatory neurons, and the marker glutamate decarboxylase, GAD, the product of the GAD1 and GAD2 genes, was used to specifically identify inhibitory neurons. The results of double staining for cell markers and c-Fos protein showed that during retrieval of associative aversive context memory in the CA1 region of the hippocampus, both Emx1-positive excitatory neurons and, less, GAD-positive inhibitory interneurons were activated. At the same time, regardless of the type of behavioral procedure (retrieval of associative context memory, non-associative context memory, or exploration of context, where animals previously received the footshock but did not remember it), the proportion of activated excitatory and inhibitory neurons remained constant, only the number of activated cells of each type changed. Altogether, our results indicate the specific role of hippocampal CA1 neurons in associative context memory and demonstrate that both excitatory and inhibitory neurons are involved in the encoding of such memory.