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A detailed proteomic analysis of plasma membrane from zebrafish brain

ABSTRACT: The brain plasma membrane proteome represents the major subsets of signaling proteins and promising drug targets though often understudied due to traditional experimental difficulties. Here we report a comprehensive dataset of the proteins identified in the highly enriched plasma membrane of zebrafish brain by applying multidimensional liquid chromatography coupled with tandem-mass spectrometry (MS/MS). A total number of 108767 peptide groups were identified resulting in 9201 proteins. These included brain receptors, transporters and channels that are important elements linked to brain functions, diseases and drug discovery.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Danio Rerio (zebrafish) (brachydanio Rerio)

TISSUE(S): Brain

SUBMITTER: Roman Smidak

LAB HEAD: Prof. Gert Lubec

PROVIDER: PXD004209 | Pride | 2018-07-05

REPOSITORIES: Pride

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A detailed proteomic profiling of plasma membrane from zebrafish brain.

Smidak Roman R Aradska Jana J Kirchberger Stefanie S Distel Martin M Sialana Fernando J FJ Wackerlig Judith J Mechtcheriakova Diana D Lubec Gert G Lubec Gert G

Proteomics. Clinical applications 20160812 12

Zebrafish (Danio rerio) is a well-established model organism in developmental biology and disease modeling. In recent years, an increasing amount of studies used zebrafish to analyze the genetic changes underlying various neurological disorders. The brain plasma membrane proteome represents the major subsets of signaling proteins and promising drug targets, but is often understudied due to traditional experimental difficulties including problems with solubility, detergent removal, or low abundan ...[more]

PMID: 27459904

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Project description:Background: Diclofenac is a non-steroidal anti-inflammatory drug frequently found in the aquatic environment. Previous studies have reported histological changes in the liver, kidney and gills of fish at concentrations similar to those measured in treated sewage effluents (~1 µg/L). Analyses or predictions of blood plasma levels in fish allow a direct comparison with human therapeutic plasma levels, and may therefore be used to indicate a risk for pharmacological effects in fish. To relate internal exposure to a pharmacological interaction we therefore investigated global hepatic gene expression together with bioconcentration to blood plasma and liver of rainbow trout (Oncorhynchus mykiss) exposed to waterborne diclofenac. Results: At the highest exposure (81.5µg/L) the fish plasma concentration reached ~88% of the human therapeutic levels (Cmax) after two weeks. Using an oligonucleotide microarray followed by quantitative PCR we found extensive effects on hepatic gene expression at this concentration, and some genes were regulated down to the lowest concentration tested (1.6 µg/L) corresponding to ~1.5% of the human Cmax. Functional analysis of differentially expressed genes revealed effects on biological processes such as inflammation and immune response, in agreement with the expected mode of action of diclofenac. In contrast to some previously reported results, the bioconcentration factor was found to be stable (4.02±0.75 for blood plasma and 2.54±0.36 for liver) regardless of the water concentration. Conclusions: Hepatic gene expression is affected in fish at blood levels similar to and below the human Cmax. This adds confidence to the use of fish blood plasma levels to predict risks for pharmacological responses in fish. The microarray results indicate similarities in the mode of action of diclofenac between humans and fish. The identified set of regulated genes may contain useful exposure biomarkers to be used in complex exposure scenarios. Juvenile rainbow trout was exposed to waterborne diclofenac at a series of different concentrations for 14 days. A flow-through system was used for eight aquaria (two per concentration), each containing 12 trouts. Nominal exposure concentrations: 1µg/L, 10 µg/L, 100 µg/L and control. Total RNA for biotinylated aRNA synthesis was extracted from the liver. Samples of aRNA from two fish from the same aquarium were pooled before the assay. In total, four biochips were analyzed corresponding to 32 microarrays. As we used two aquaria per concentration, these 32 microarrays allowed the analyses of four aRNA pools (eight fish) per aquarium, which is equal to eight pools per concentration of diclofenac.

			Action	DRS
	Chymotrypsin.msf	Msf
	Identifications.pdResult	Other
	NP11-TDr-BA1-O1005_0-1.raw	Raw
	NP11-TDr-BA1-O1005_0-2.raw	Raw
	NP11-TDr-BA2-O1005_1-1.raw	Raw