Reference genes to quantify gene expression during oogenesis in a teleost fish. | - CCMAR -

Journal Article

TitleReference genes to quantify gene expression during oogenesis in a teleost fish.
Publication TypeJournal Article
AuthorsDeloffre, LAM, Andrade, A, Filipe, AI, Canario, AVM
Year of Publication2012
Date Published2012 Sep 10
KeywordsActins, Animals, Base Sequence, Cathepsin D, Cathepsin Z, DNA Primers, Female, Fish Proteins, Gene Expression, Genetic Markers, Oogenesis, Peptide Elongation Factor 1, Real-Time Polymerase Chain Reaction, TATA-Box Binding Protein, Tilapia, Tubulin

Understanding the molecular events involved in the acquisition of competence during oogenesis is a key step to determine the secret of 'high quality' eggs for aquaculture. Quantitative real time polymerase chain reaction (qPCR) is the technique of election to determine changes in transcript abundance in such studies, but choosing reference genes for normalization, in particular during oogenesis, remains a challenge. In the present study, transcription of 6 functionally distinct genes, β actin (ACTB), cathepsin D (CTSD), cathepsin Z (CTSZ), elongation factor 1 α (EEF1A), TATA binding protein (TBP) and tubulin A (TUBA1A) was assessed as normalizers of bone morphogenetic protein (BMP) and activin membrane-bound inhibitor (BAMBI) gene expression in mRNA from Mozambique tilapia oocytes during oogenesis. Reverse transcription was equally efficient and varies little in all samples. Most of the genes considered for reference were stable during early stages of oogenesis but variations were observed during vitellogenesis. A single gene and up to 3 genes were shown to be insufficient for reliable normalization throughout the whole oogenesis. The combination of the genes ACTB, CTSD, EEF1A and CTSZ as reference was found to minimize variation and has the most stable expression pattern between maturation stages.


Alternate JournalGene
PubMed ID22750322
CCMAR Authors