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Using skin mucus RNA to study gene expression in pacific bluefin tuna
Author(s) -
Castilho Pedro Carvalho,
Jayasundara Nishad,
Mladineo Ivona,
Block Barbara B
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.187
Subject(s) - tuna , biology , complementary dna , mucus , rna extraction , rna , gene expression , agarose gel electrophoresis , fishery , reverse transcriptase , atlantic cod , gene , gadus , genetics , ecology , fish <actinopterygii>
The bluefin tuna species have been overfished throughout the world with stocks declining significantly over the past two decades. Studies employing electronic tags describe tunas in extreme ocean environments with depths up to 500 meters and water temperatures ranging from 5 to 33°C. Physiological adaptations such as high cardiac output and endothermy lead tunas to be a good model for studies in cardiac physiology and stress response. However, due to the high cost of tuna maintenance in captivity, studies on tuna physiology and gene expression are rare. Here, we described the use of pacific bluefin tuna ( Thunnus orientalis ) skin mucus as a possible source for RNA and its potential to be used in gene expression studies of fishes. The skin mucus samples were taken from tunas in captivity at the Tuna Research and Conservation Center and stored at −20°C in RNAlater tubes (Invitrogen). Total RNA extraction from the mucus samples were performed using RNAeasy Kit (Qiagen), posterior RNA clean‐up was performed using Ambion Filter Cartridge. The RNA concentration and quality was spectrophotometrically analyzed by A 260 /A 280 ratio and from agarose gel electrophoresis. Reverse transcription of RNA from mucus samples into cDNA was performed using iScript cDNA kit (Bio‐Rad). PCR products from mucus cDNA samples, were successfully amplified using bluefin tuna gene specific primers (Hsp70; β‐actin; IL‐1β). Our results indicate that bluefin tuna skin mucus is a potential source of RNA for gene expression studies in fishes. Funding: NOAA, MBA.