GSM4340764: CGSMG_Snap25_ip_GF_1; Mus musculus; OTHER - SRA

SRX7806874: GSM4340764: CGSMG_Snap25_ip_GF_1; Mus musculus; OTHER
1 ILLUMINA (NextSeq 500) run: 26.6M spots, 2G bases, 727.8Mb downloads

Submitted by: NCBI (GEO)

Study: Microbes modulate sympathetic neurons via a gut-brain circuit

PRJNA608916 • SRP250834 • All experiments • All runs

show Abstracthide Abstract

Gut-brain connections monitor the intestinal tissue and its microbial and dietary content1, regulating both intestinal physiological functions such as nutrient absorption and motility2,3, and brain–wired feeding behaviour2. It is therefore plausible that circuits exist to detect gut microbes and relay this information to central nervous system (CNS) areas that, in turn, regulate gut physiology4. We characterized the influence of the microbiota on enteric–associated neurons (EAN) by combining gnotobiotic mouse models with transcriptomics, circuit–tracing methods, and functional manipulation. We found that the gut microbiome modulates gut–extrinsic sympathetic neurons; while microbiota depletion led to increased cFos expression, colonization of germ-free mice with short-chain fatty acid–producing bacteria suppressed cFos expression in the gut sympathetic ganglia. Chemogenetic manipulations, translational profiling, and anterograde tracing identified a subset of distal intestine-projecting vagal neurons positioned to play an afferent role in microbiota–mediated modulation of gut sympathetic neurons. Retrograde polysynaptic neuronal tracing from the intestinal wall identified brainstem sensory nuclei activated during microbial depletion, as well as efferent sympathetic premotor glutamatergic neurons that regulate gastrointestinal transit. These results reveal microbiota–dependent control of gut extrinsic sympathetic activation through a gut-brain circuit. Overall design: Actively translated mRNA profiles from immunoprecipitated ribosome-bound mRNA from the nodose ganglion, dorsal root ganglion, and celiac superior mesentetic of Snap25:RiboTag specific pathogen free or germ free mice were prepared by deep sequencing on an Illumina NextSeq. Actively translated mRNA profiles from immunoprecipitated ribosome-bound mRNA from the nodose ganglion of SNS:RiboTag, Nav1.8:RiboTag, and Advillin:RiboTag mice were prepared by deep sequencing on an Illumina NextSeq 16S RNA profiles, from cecal samples of C57BL6/J mice given streptomycin or PBS, were prepared for deep sequencing on an Illumina MiSeq.

Sample: CGSMG_Snap25_ip_GF_1

SAMN14213180 • SRS6219909 • All experiments • All runs

Organism: Mus musculus

Library:

Instrument: NextSeq 500

Strategy: OTHER

Source: TRANSCRIPTOMIC

Selection: other

Layout: SINGLE

Construction protocol: The nodose ganglion, dorsal root ganglion, and celiac superior mesenteric ganglion were dissected into PBS with cycloheximide (0.2 mg/mL) (PBS/CHX). The RiboTag immunoprecipitation protocol (http://depts.washington.edu/mcklab/RiboTagIPprotocol2014.pdf) was then followed with the following modifications: All samples were homogenized by hand with a dounce homogenizer in 2.5 mL supplemented homogenization buffer (changes per 2.5 mL: 50 µl Protease Inhibitor, 75 µl heparin (100 mg/mL stock), 25 µl SUPERase In RNase Inhibitor). Samples were then centrifuged for 10 minutes at 10,000 G, after which 800 µl of supernatant was removed and 5µL of anti-HA antibody (Abcam, ab9110) was added. For input samples, 40uL of supernatant was removed and 60uL of PicoPure lysis buffer was added. These input samples were then put at -80°C. Immunoprecipitated samples were kept rotating at 4°C with antibody for 1 hour. 200 µl of Thermo Protein magnetic A/G beads were washed with homogenization buffer, added to the sample, and kept rotating for 30 minutes at 4°C. The beads were washed four times with high-salt buffer and samples were eluted with 100 µl of PicoPure lysis buffer. RNA was extracted using the Arcturus PicoPure RNA isolation kit (Applied Biosystems) according to the manufacturer's instructions.

Experiment attributes:

GEO Accession: GSM4340764

Links:

NCBI link: NCBI Entrez (gds)

Runs: 1 run, 26.6M spots, 2G bases, 727.8Mb

Run	# of Spots	# of Bases	Size	Published
SRR11186590	26,573,015	2G	727.8Mb	2020-03-22

ID:: 10211485

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