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Identifier

000364009

Title

Μελέτη της έκφρασης των δύο ισομορφών του γονιδίου της Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) σε αναπτυξιακά στάδια και ιστούς του λαυρακιού (Dicentrarchus labrax) και της τσιπούρας (Sparus aurata)

Alternative Title

Study of the expression of the two GAPDH isoforms in different developmental stages and tissues of the European seabass (Dicentrarchus labrax) and the gilthead seabream (Sparus aurata)

Author

Νουσδίλη, Δήμητρα

Thesis advisor

Λαδουκάκης, Μανόλης
Κωτούλας, Γεώργιος
Κεντούρη, Μαρουδιώ

Abstract

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is of principal role in basic cell catabolic processes and in the pathway of energy. It was considered as a housekeeping gene due to its stable expression at high levels in most tissues and cells under all conditions. For this reason, GAPDH is commonly used as an internal control for RT-PCR studies. However, recent evidence for differential expression patterns in different organisms and tissues as well as the occurrence of different isoforms and pesudogenes, driving to the result that it is necessary to reassess its role of internal control. In addition to this, GAPDH is involved in transcription, apoptosis and several other functions. In the present study, we isolated the two isoforms GAPDH 1 and GAPDH 2 of the gilthead sea bream (Sparus aurata) and the European sea bass (Dicentrarchus labrax) and described the way of the identification and characterization. We performed an in silico phylogenetic analysis of both GAPDH paralogues in 10 fish species, including sea bream and sea bass, as well as test of selection. No positive selection was detected. In contrast, negative selection was detected. Moreover, we studied the gene structure of both isoforms and the syntenic relationships among fish species with known genomes. We further investigated the gene expression patterns of both isoforms in sea bream and sea bass in different developmental stages and tissues using RT-qPCR method and One-step RT-PCR respectively. The selected range of developmental stages for 9 sea bass was from the morula stage to the hatched stage. As for sea bream was from the gastrula stage to the stage of eye pigmentation as well as further stages of early development (from two cell stage to gastrula), showing divergent gene expression of the two isoforms and suggesting the possible function of GAPDH 1 as a maternal gene. Further investigation is needed for the tissue-specificity of GAPDH isoforms.

Language

Greek

Subject

Gene expression

Ισομορφή

Λαυράκι

Τριπούρα

Issue date

2011-03-18

Collection

School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses

Type of Work--Post-graduate theses