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Transcriptional profiling of neurons in the rostral pons of a Cbln2-TRAP (NIDA172) mouse line

ABSTRACT: Transcriptional profiling of neurons in the rostral pons of a Cbln2-TRAP (NIDA172) mouse line

PROVIDER: PRJNA563059 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10051035_1.fastq.gz	Fastqsanger.gz
	SRR10051035_2.fastq.gz	Fastqsanger.gz
	SRR10051036_1.fastq.gz	Fastqsanger.gz
	SRR10051036_2.fastq.gz	Fastqsanger.gz
	SRR10051037_1.fastq.gz	Fastqsanger.gz

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Transcriptional profiling of neurons in the rostral pons of a Cbln2-TRAP (NIDA172) mouse line

Project description:TRAP translational profiling is a method that allows investigators to genetically characterize specific cell types in complex tissues such as mouse brain. Using this technique we obtained RNA-Seq data from actively translating transcripts present in neurons in the rostral pons of adult Cbln2-EGFP/Rpl10a (NIDA172) mice. This brain area includes the locus coeruleus and parabrachial nuclei. This work was supported by a grant from the National Institute on Drug Abuse P30DA035756 to N.H.

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Project description:The 5HT system is organized into rostral and caudal populations with discrete anatomical locations and opposite axonal trajectories in the developing hindbrain. 5HT neuron cell bodies in the rostral subdivision migrate to the midbrain and pons and extend ascending projections throughout the forebrain. 5HT cell bodies in the caudal subdivision migrate to the ventral medulla and caudal half of the pons and provide descending projections to the brainstem and spinal cord. Experiment Overall Design: We used microarrays to determine genes expressed by both rostral and caudal 5HT neurons as well as genes that are differentially expressed between rostral and caudal 5HT neurons at E12.5 when axon pathfinding and cell migration are underway. E12.5 neural tubes were isolated from ePet-EYFP embryos and dissected into a rostral domain (mesecephalic flexure to pontine flexure) and a caudal domain (pontine flexure to spinal cord). After cell dissociation (details under growth protocol), cells were subjected to fluorescent activated cell sorting (FACS) to obtain 4 cell populations. R+ = rostral ePetEYFP positive 5HT neurons; R- = YFP negative non-serotonergic cells in the rostral neural tube; C+ = caudal ePetEYFP positive 5HT neurons; C- = YFP negative non-serotonergic cells in the caudal neural tube. 200,000 cells for each of the 4 cell types (R+, R-, C+, C-) were collected for RNA extraction and hybridization to Affymetrix Mouse 430 2.0 arrays.

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