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dc.contributor.advisorCramb, Gordon
dc.contributor.advisorHazon, N. (Neil)
dc.contributor.authorMcWilliam, Iain Stuart
dc.descriptionElectronic version does not contain associated previously published materialen
dc.description.abstractThe European eel, Anguilla anguilla, has a complex life-cycle involving migrations between the Sargasso Sea and the river systems of Europe and North Africa. The requirement to move across large salinity gradients presents a significant physiological challenge and the developmental stages of the eel are closely linked to these migrations. Microarrays were created to elucidate gene expression changes occurring during; i. The transition from juvenile yellow to the adult sexually maturing, migrating silver eel and; ii. Salinity adaptation during the migration from freshwater to seawater. Groups (n = 6) of freshwater-acclimated yellow or silver eels were transferred to seawater for between 6 hours and 5 months and complementary control groups were transferred to freshwater. Brain, kidney, intestine and gill cDNA libraries were constructed using suppression subtractive hybridisation (SSH) techniques and a novel protocol based on Invitrogen's Gateway cloning system. The latter technique produced a low redundancy (~4 %) EST bank with a wide range of insert sizes (0.5 – 10 kb). Two microarray types were produced; one comprised 5760 clones from the two brain libraries whilst the other was a multi-tissue microarray incorporating 6144 clones from the SSH libraries. Pooled RNA samples were probed against the microarrays to highlight differentially expressed genes. Real-time quantitative PCR (QPCR) was used to validate the observed expression changes of selected genes in the tissues of individual fish. Following yellow to silver transformation of freshwater-adapted eels, the expression of tyrosine 3-mono-oxygenase/tryptophan 5-mono-oxygenase activation protein (14-3-3) and vaccinia related kinase 3 was shown to be consistently elevated. Prolactin expression increased in the brains of silver eels following two-day seawater-acclimation but QPCR analysis revealed high variation amongst freshwater-adapted eels. This is the first eel microarray study and the expression profiles highlighted herein will provide new avenues for research into the sexual development and salinity acclimation of A. anguillaen
dc.format.extent13329712 bytes
dc.publisherUniversity of St Andrews
dc.rightsCreative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported
dc.subjectEuropean eelen
dc.subjectcDNA libraryen
dc.subjectGene expressionen
dc.subjectSexual developmenten
dc.subject.lcshDNA microarraysen
dc.subject.lcshAnguilla anguilla--Geneticsen
dc.subject.lcshAnguilla anguilla--Developmenten
dc.subject.lcshGene expressionen
dc.titleDevelopment of microarray techniques for the study of gene expression in the European eel (Anguilla anguilla) during silvering and migration to seawateren
dc.contributor.sponsorNatural Environment Research Council (NERC)en
dc.type.qualificationnamePhD Doctor of Philosophyen
dc.publisher.institutionThe University of St Andrewsen

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Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported
Except where otherwise noted within the work, this item's licence for re-use is described as Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported