Central amygdala cell types and their transcriptional regulation by fasting

A complete cell type census based on their full molecular profiles at subclass level and their subnuclei distribution in the central amygdala (CeA), linked with their behavior relevance, is still lacking. Here, we performed single nuclei RNA sequencing in adult mice and compared the results with known genetic and functionally defined populations. We identified 9 transcriptionally and spatially discrete cell types (3 in the lateral division CeL; 5 in the medial division CeM; and 1 in the capsular division CeC). In the CeL, we found one PKCd expressing cluster (CeL PKCd ) and two clusters marked by Sst: the CeL Sst cluster which contains the largest and most highly expressing population of Sst-positive neurons, and the CeL NTS/Tac2 cluster which contains low expressing Sst-positive cells. This two Sst population are also Htr2a positive. The CeC contains one cluster marked by the expression of Calcrl and partially expressed PKCd. In the CeM, the largest cluster was marked by Il1Rapl2, which contains the CeM subsets of Pnoc and Htr2a, and is likely appetitive. Another Sst population, the CeM Tac1/Sst, remains largely unexplored. Moreover, we discovered three previously uncharacterized CeM clusters: CeM Vdr, CeM Drd2/Rai14, and CeM Dlk1. To begin investigating whether these cell types show individual responses in appetitive behavior, we compared their transcriptomes in food-deprived and satiated mice. We found a cell type specific transcriptional response across neuronal populations, particularly in one of the appetitive clusters in the CeL. Overall design: Mouse CeA were dissected out and single nuclei suspension were prepared. Two data sets (biological replicate) were obtained from 8-week old naïve mice. A third data set came from Htr2a-Cre transgenic mice expressing AAV-DIO-mCherry to determine the CeAHtr2a population. Another two datasets (biological replicate) were obtained from mice that were fasted for 20 hours.