GNPS_GC_3D_skin_mapping_longitudinal_high_res_min
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Agilent 7200 GC-QTOF
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: 
Pieter Dorrestein
PROVIDER: MSV000084338 | GNPS |
REPOSITORIES: GNPS
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Project description:All samples were collected on cotton then placed into 2 mL borosilicate vials. The GC MS analysis was carried out using the Agilent 7200 GC QTOF Agilent Technologies Santa Clara CA. equipped with a robotic sampler system. The separation was conducted on an HP 5MS column (30 m x 0.25 mm x 0.25 micro m). The patch within the vial was heated for 25 min. at 200 C to desorb volatiles from the patch and 0.5 mL of headspace injected (injector temperature set at 250 C) into the instrument with a headspace syringe heated to 145 C. The GC protocol analysis included starting temperature 45 C min oven ramp (hold of 2 min), 15 C per min oven ramp to 300 C (hold of 3 min), and 50 C per min oven ramp to 320 C purge the column. The helium carrier gas was set to constant 1.2 mL per min flow and a spitless injection mode was applied. The purge flow to split vent rate was 50 ml per min at 1 min. The scanned m z range was 35-400 with the acquisition rate of 10 spectra per s. The empty vial blanks were interspersed with the samples to assess the background signal.
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Project description:Twenty transgenic fish overexpressing agouti-signaling protein (ASIP, Ceinos et al., Pigment Cell and Melanoma Research 28:196-209, 2015) and wild type (WT) zebrafish were placed in 6-liter tanks and acclimated during 30 days prior to sampling to minimize physiological differences. Animals were sacrificed and the brain+pituitary and the skin dissected under the stereo microscope, placed in trizol and stored at -80C until RNA extraction. Six biological replicates (3 females and 3 males) of each tissue (skin and brain) of each group (ASIP vs WT) were analyzed in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Global gene expression analyses were performed to search for both sex-dependent and transgene-dependent effects as well as the interaction between both factors (sex x phenotype) on the brain+pituitary and skin transcriptome. Wild type (Tu) or transgenic zebrafish overexpression agouti-signaling protein (ASIP) were reared in similar conditions. Brain+pituitary and skin were sampled from 6 ASIP (3 males + 3 females) and WT (3 males + 3 females) zebrafish. Samples were individually hybridized in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Microarrays included 60-mer probes to the unique transcripts from Ensembl and Unigene which were annotated by functional categories of GO and pathways of KEGG with bioinformatics package STARS
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