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Single cell RNA sequencing revealing gene expression profile of thalamus from mice at the age of 2 months

ABSTRACT: Single cell RNA sequencing revealing gene expression profile of thalamus from mice at the age of 2 months

PROVIDER: PRJNA1124372 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29424192_1.fastq.gz	Fastqsanger.gz
	SRR29424192_2.fastq.gz	Fastqsanger.gz
	SRR29424193_1.fastq.gz	Fastqsanger.gz
	SRR29424193_2.fastq.gz	Fastqsanger.gz
	SRR29424194_1.fastq.gz	Fastqsanger.gz

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Project description:The thalamus may be the critical brain area involved in sensory gating and the relay of respiratory mechanical information to the cerebral cortex for the conscious awareness of breathing. We hypothesized that respiratory mechanical stimuli in the form of tracheal occlusions would modulate the gene expression profile of the thalamus. Specifically, it was reasoned that conditioning to the respiratory loading would induce a state change in the medial thalamus consistent with a change in sensory gating and the activation of molecular pathways associated with learning and memory. In addition, respiratory loading is stressful and thus should elicit changes in gene expressions related to stress, anxiety, and depression. Rats were instrumented with inflatable tracheal cuffs. Following surgical recovery, they underwent ten days (5 days/week) of transient tracheal occlusion conditioning. On day 10, the animals were sacrificed and the brains removed. The medial thalamus was dissected and microarray analysis of gene expression performed. Tracheal obstruction conditioning modulated a total of 661 genes (p < 0.05, log2 fold change ≥ 0.58), 250 genes were down-regulated and 411 up-regulated. There was a significant down-regulation of GAD1, GAD2 and HTR1A, HTR2A genes. CCK, PRKCG, mGluR4, and KCJN9 genes were significantly up-regulated. Some of these genes have been associated with anxiety and depression, while others have been shown to play a role in switching between tonic and burst firing modes in the thalamus and thus may be involved in gating of the respiratory stimuli. Furthermore, gene ontology and pathway analysis showed a significant modulation of learning and memory pathways. These results support the hypothesis that the medial thalamus is involved in the respiratory sensory neural pathway due to the state change of its gene expression profile following repeated tracheal occlusions. Four experimental animals that received tracheal occlusions and four surgical control animals were used. A reference design was used to compare all samples.

Project description:Conscious awareness of breathing requires the activation of higher brain centers. The sensation of breathing is believed to be a neural gating process. The thalamus provides the brain structure that could be responsible for the gating of respiratory sensory information to the cortex. It was reasoned that if the thalamus is the neural gate, then tracheal obstructions will modulate the gene expression profile of the thalamus. Anesthetized rats were instrumented with an inflatable cuff sutured around the trachea. The cuff was inflated to obstruct 2-4 breaths, then deflated for a minimum of 15 breaths. The obstructions were repeated for 10 min followed by immediate brain removal, and the medial thalamus was dissected and prepared for microarray analysis. Gene expression profiles were measured using Agilent Technology Oligo Microarrays. Following the occlusion protocol, 588 genes were found to be altered (p < 0.05, log2 fold change ≥ 0.4), with 327 down-regulated and 261 genes up-regulated. A significant up-regulation of the serotonin HTR2A receptor and significant down-regulation of the dopamine DRD1 receptor genes were found. A pathway analysis was performed targeting serotonin and dopamine receptor pathways. The mitogen activated protein kinase 1 (MAPK1) gene was significantly down-regulated. MAPK1 is an inhibitory regulator of the serotonin HTR2A receptor and facilitatory regulator for the dopamine DRD1 receptor. Down-regulation of MAPK1 may be related to the 2-fold up-regulation of HTR2A and 2-fold down-regulation of DRD1 suggesting an interaction in the medial thalamus serotonin-dopamine pathway elicited by airway obstruction. These results demonstrate an immediate change in gene expression in thalamic arousal-fear-anxiety-motivation related serotonin and dopamine receptors in response to airway obstruction. The results support the hypothesis that the thalamus is a component in the respiratory mechanosensory neural pathway. Four experimental animals that received tracheal occlusions and four surgical control animals were used. A reference design was used to compare all samples.

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Single cell RNA sequencing revealing gene expression profile of thalamus from mice at the age of 2 months

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