Premium
Analysis of transcriptional responses of normalizing genes on Crassostrea brasiliana under different experimental conditions
Author(s) -
Müller Gabrielle do Amaral e Silva,
de Lima Daína,
Zacchi Flávia Lucena,
Piazza Rômi Sharon,
Lüchmann Karim Hahn,
Mattos Jacó Joaquim,
Schlenk Daniel,
Bainy Afonso Celso Dias
Publication year - 2017
Publication title -
environmental toxicology and chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.1
H-Index - 171
eISSN - 1552-8618
pISSN - 0730-7268
DOI - 10.1002/etc.3755
Subject(s) - reference genes , biology , gene , gene expression , ribosomal rna , transcriptome , microbiology and biotechnology , biochemistry
Bivalves show remarkable plasticity to environmental changes and have been proposed as sentinel organisms in biomonitoring. Studies related to transcriptional analysis using quantitative real‐time polymerase chain reaction (qRT‐PCR) in these organisms have notably increased, imposing a need to identify and validate adequate reference genes for an accurate and reliable analysis. In the present study, 9 reference genes were selected from transcriptome data of Crassostrea brasiliana to identify their suitability as qRT‐PCR normalizer genes. The transcriptional patterns were analyzed in gills of oysters under 3 different conditions: different temperatures (18, 24, or 32 °C) and phenanthrene (100 µg L −1 ) combined exposure; different salinities (10, 25, or 35‰) and phenanthrene combined exposure; and 10% of diesel fuel water‐accommodated fraction (diesel‐WAF) exposure. Reference gene stability was calculated using 5 algorithms (geNorm, NormFinder, BestKeeper, ΔCt, RefFinder). Transcripts of ankyrin ‐ like ( ANK ), glyceraldehyde 3‐phosphate dehydrogenase‐like (GAPDH), and α‐tubulin ‐ like ( TUBA ) genes showed minor changes in different temperature/phenanthrene treatment. Transcripts of ANK, β‐actin‐like , and β‐tubulin‐like genes showed better stability at salinity/phenanthrene treatment, and ANK, TUBA , and 28S ribosomal protein‐like genes showed the most stable transcription pattern in oysters exposed to diesel‐WAF exposure. The present study constitutes the first systematic analysis of reference gene selection for qRT‐PCR normalization in C. brasiliana . These genes could be employed in studies using qRT‐PCR analysis under similar experimental conditions. Environ Toxicol Chem 2017;36:2190–2198. © 2017 SETAC