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The cellular basis of distinct thirst modalities

ABSTRACT: The cellular basis of distinct thirst modalities

PROVIDER: PRJNA644927 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12179476.fastq.gz	Fastqsanger.gz
	SRR12179477.fastq.gz	Fastqsanger.gz
	SRR12179478.fastq.gz	Fastqsanger.gz
	SRR12179479.fastq.gz	Fastqsanger.gz
	SRR12179480.fastq.gz	Fastqsanger.gz

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The cellular basis of distinct thirst modalities

Project description: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.

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The cellular basis of distinct thirst modalities.

Project description:Fluid intake is an essential innate behaviour that is mainly caused by two distinct types of thirst1-3. Increased blood osmolality induces osmotic thirst that drives animals to consume pure water. Conversely, the loss of body fluid induces hypovolaemic thirst, in which animals seek both water and minerals (salts) to recover blood volume. Circumventricular organs in the lamina terminalis are critical sites for sensing both types of thirst-inducing stimulus4-6. However, how different thirst modalities are encoded in the brain remains unknown. Here we employed stimulus-to-cell-type mapping using single-cell RNA sequencing to identify the cellular substrates that underlie distinct types of thirst. These studies revealed diverse types of excitatory and inhibitory neuron in each circumventricular organ structure. We show that unique combinations of these neuron types are activated under osmotic and hypovolaemic stresses. These results elucidate the cellular logic that underlies distinct thirst modalities. Furthermore, optogenetic gain of function in thirst-modality-specific cell types recapitulated water-specific and non-specific fluid appetite caused by the two distinct dipsogenic stimuli. Together, these results show that thirst is a multimodal physiological state, and that different thirst states are mediated by specific neuron types in the mammalian brain.

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