Abstract
Fat metabolism in toothed whales is different from other mammals.
RT-qPCR is still a reliable technique for studying the relative
expressions of various genes involved in metabolism. This study was done
for Risso’s dolphin, a toothed whale and information produced here will
be important for further transcriptomics studies focused on unrevealed
marine mammal fat metabolism. In this study, we sought to identify a
suitable reference gene with minimum resources. Seven candidate
reference genes ZC3H10, FTL, LGALS1, RPL27A, GAPDH, FTH1 and DCN were
initially tested for amplification efficiency using RT-qPCR by producing
standard curves. Then, three nearly 100% efficient genes FTL, LGALS1
and GAPDH were selected for the gene stability analysis to determine one
stable gene across eight different fat tissues, liver, and muscle of
Risso’s dolphins based on four algorithms, provided in geNorm,
NormFinder, BestKeeper and Delta Ct. Finally, a RefFinder comprehensive
ranking was done based on stability values and the genes were ranked as:
FTL>LGALS1>GAPDH. The FTL and LGHLS were
identified as the most stable genes; however, GAPDH was third, a
well-known housekeeping gene for mammals. Finally, we suggest using FTL
as a reliable reference gene for functional genomics studies on toothed
whales in the future.