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Cell type-specific RNA-seq to profile transcriptional changes in sorted BLA projection neurons during conditioned taste aversion learning

ABSTRACT: Cell type-specific RNA-seq to profile transcriptional changes in sorted BLA projection neurons during conditioned taste aversion learning

PROVIDER: PRJNA576246 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10240003.fastq.gz	Fastqsanger.gz
	SRR10240004.fastq.gz	Fastqsanger.gz
	SRR10240005.fastq.gz	Fastqsanger.gz
	SRR10240006.fastq.gz	Fastqsanger.gz
	SRR10240007.fastq.gz	Fastqsanger.gz

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Cell type-specific RNA-seq to profile transcriptional changes in sorted BLA projection neurons during conditioned taste aversion learning

Project description:In order to identify specific transcripts that might be necessary for CTA learning within the BLA, we used cell type-specific RNA-seq to profile transcriptional changes in sorted BLA projection neurons 4 hours following training.

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Project description:Animals must learn through experience which foods are nutritious and should be consumed, and which are toxic and should be avoided. Enteroendocrine cells (EECs) are the principal chemosensors in the GI tract, but investigation of their role in behavior has been limited by the difficulty of selectively targeting these cells in vivo. Here we describe an intersectional genetic approach for manipulating EEC subtypes in behaving mice. We show that multiple EEC subtypes inhibit food intake but have different effects on learning. Conditioned flavor preference is driven by release of cholecystokinin whereas conditioned taste aversion is mediated by serotonin and substance P. These positive and negative valence signals are transmitted by vagal and spinal afferents, respectively. These findings establish a cellular basis for how chemosensing in the gut drives learning about food.

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Project description:Feeding is an important activity for all animals providing nutrients essential for survival and reproduction. Not surprisingly, learning plays a critical role in feeding behavior through the establishment and strengthening of food preferences and aversions. That is, the integration of taste and post ingestive visceral signals in the brain results in memorial representations about the consequences associated with ingesting a particular food. For example, when ingestion of a food is followed by negative gastrointestinal consequences (e.g. nausea, sickness, or vomiting), the animal develops a conditioned taste aversion (CTA), which produces a switch from acceptance to avoidance of that and any like tasting stimulus. Despite recent advances in understanding CTA responsive intracellular signaling pathways in the amygdala, little is known about any long-term regulation of target gene expression following CTA memory consolidation and retrieval. The present study utilized oligo-nucleotide microarray to understand the genes and networks involved in Conditional Taste Aversion Behavior.

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Project description:The present experiments investigated gene expression in the amygdala following contingent taste/LiCl treatment that supports development of conditioned taste aversion (CTA). The use of whole genome chips and stringent data set filtering led to the identification of 168 genes regulated by CTA compared to non-contingent LiCl treatment that does not support CTA learning. Seventy-six of these genes were eligible for network analysis. Such analysis identified "behavior" as the top biological function, which was represented by 15 of the 76 genes. These genes included several neuropeptides, G protein-coupled receptors, ion channels, kinases, and phosphatases. Subsequent qRT-PCR analyses confirmed changes in mRNA expression for 5 of 7 selected genes. We were able to demonstrate directionally consistent changes in protein level for 3 of these genes; insulin 1, oxytocin, and major histocompatibility complex class I-C. Behavioral analyses demonstrated that blockade of central insulin receptors produced a weaker CTA that was less resistant to extinction. Together, these results support the notion that we have identified downstream genes in the amygdala that contribute to CTA learning.

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Project description:Two independent lines of High Drinking in the Dark (HDID-1, HDID-2) mice have been bred to reach high blood alcohol levels after a short period of binge-like ethanol drinking. Male mice of both lines were shown to have reduced sensitivity to develop a taste aversion to a novel flavor conditioned by ethanol injections as compared with their unselected HS/NPT founder stock. We have subsequently developed inbred variants of each line. The current experiments established that reduced ethanol-conditioned taste aversion is also seen in the inbred variants, in both males and females. In other experiments, we asked whether HDID mice would ingest sufficient doses of ethanol to lead to a conditioned taste aversion upon retest. Different manipulations were used to elevate consumption of ethanol on initial exposure. Access to increased ethanol concentrations, to multiple tubes of ethanol, and fluid restriction to increase thirst motivation all enhanced initial drinking of ethanol. Each condition led to reduced intake the next day, consistent with a mild conditioned taste aversion. These experiments support the conclusion that one reason contributing to the willingness of HDID mice to drink to the point of intoxication is a genetic insensitivity to the aversive effects of ethanol.

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