We performed single-cell RNA-seq (10x Chromium 3' v.2) on dissociated single cell suspensions from mouse (C57BL/6) osmosensory brain nuclei - subfornical organ (SFO) and organum vasculosum lamina terminalis (OVLT) to determine transcriptomic cell types in each structure. We identified 12 and 13 major cell classes as well as 8 neuron types in each for SFO and OVLT respectively. Furthermore, we studied how different types of thirst signals (osmotic thirst, hypovolemic thirst, chronic water deprivation) engage this cellular diversity by performing single cell RNA-seq based (10x Chromium 3' v.3.0) stimulus to cell-type mapping, where dissociation related transcriptional artifacts were suppressed through a modified tissue preparation protocol involving a transcriptional blocker and dissociation at room temperature. This approach revealed the endogenous immediate early gene expression triggered by distinct thirst stimuli in SFO and OVLT cells. By analyzing immediate early gene expression that is driven by changes in neural activity (Fos) we identified excitatory neuron types in SFO and OVLT that are uniquely tuned to distinct thirst states. SOURCE: Allan-Hermann Pool (allan.pool@gmail.com, apool@caltech.edu, yoka@caltech.edu) - Oka Lab California Institute of Technology

Pluto Bioinformatics

GSE154048: The cellular basis of distinct thirst modalities