DOI: 10.5061/dryad.4cs9573
title: Data from: Molecular physiology of chemical defenses in a poison frog

description: Poison frogs sequester small molecule lipophilic alkaloids from their diet
 of leaf litter arthropods for use as chemical defenses against predation.
 Although the dietary acquisition of chemical defenses in poison frogs is
 well-documented, the physiological mechanisms of alkaloid sequestration
 has not been investigated. Here, we used RNA sequencing and proteomics to
 determine how alkaloids impact mRNA or protein abundance in the Little
 Devil Frog (Oophaga sylvatica) and compared wild caught chemically
 defended frogs to laboratory frogs raised on an alkaloid-free diet. To
 understand how poison frogs move alkaloids from their diet to their skin
 granular glands, we focused on measuring gene expression in the
 intestines, skin, and liver. Across these tissues, we found many
 differentially expressed transcripts involved in small molecule transport
 and metabolism, as well as sodium channels and other ion pumps. We then
 used proteomic approaches to quantify plasma proteins, where we found
 several protein abundance differences between wild and laboratory frogs,
 including the amphibian neurotoxin binding protein saxiphilin. Finally,
 because many blood proteins are synthesized in the liver, we used thermal
 proteome profiling as an untargeted screen for soluble proteins that bind
 the alkaloid decahydroquinoline. Using this approach, we identified
 several candidate proteins that interact with this alkaloid, including
 saxiphilin. These transcript and protein abundance patterns suggest the
 presence of alkaloids influences frog physiology and that small molecule
 transport proteins may be involved in toxin bioaccumulation in dendrobatid
 poison frogs.
description: Oophaga sylvatica
 transcriptomeTranscriptome from
 Oophaga sylvatica. Sequenced from skin, liver, and intestines from a
 single individual. Full description of sample processing and sequencing
 can be found in associated manuscript.Osylvatica_transcriptome_Catyetal.fastaOsylvatica reference proteomeOophaga sylvatica proteome generated from mRNA-based protein reference database for peptide matching, using the PHROG tool (Proteomic Reference with Heterogeneous RNA Omitting the Genome).Osylvatica_allPHROG_proteins.faGas Chromatography Mass Spectrometry for AlkaloidsGas chromatography/mass spectrometry data from the skin of the Oophaga sylvatica. Five frogs were caught in the rainforest in Ecuador and three were laboratory raised. Dorsal skin was isolated and alkaloids were extracted with methanol.AlkaloidsGCMS.zipOophaga sylvatica Plasma Proteomics (part 1 of 3)Tandem Mass Tag (TMT; Thermo Scientific, Waltham, MA, USA) labeling was performed according to manufacturer's instructions. TMT is a chemical label that allows the quantification of proteins from pooled samples by adding slight variations to the molecular mass of proteins. Full description available in the associated manuscript.plasmaproteomics.set1.tar.gzOophaga sylvatica Plasma Proteomics (part 2 of 3)Tandem Mass Tag (TMT; Thermo Scientific, Waltham, MA, USA) labeling was performed according to manufacturer's instructions. TMT is a chemical label that allows the quantification of proteins from pooled samples by adding slight variations to the molecular mass of proteins. Full description available in the associated manuscript.plasmaproteomics.set2.tar.gzOophaga sylvatica Plasma Proteomics (part 3 of 3)Tandem Mass Tag (TMT; Thermo Scientific, Waltham, MA, USA) labeling was performed according to manufacturer's instructions. TMT is a chemical label that allows the quantification of proteins from pooled samples by adding slight variations to the molecular mass of proteins. Full description available in the associated manuscript.plasmaproteomics.set3.tar.gzOophaga sylvatica Thermal Proteome ProfilingThis assay was performed to identify proteins that bind to alkaloids. The assay relies on thermodynamic principles of protein-ligand binding, where a protein that has bound its small molecule target has increased thermal stability, a shift detectable by tandem mass spectrometry. A single laboratory reared frog was used for this analysis.TPP.tar.gz