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Title: The identification and characterisation of aminergic G-protein coupled receptors from amphioxus
Author: Burman, Chloe
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2009
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The cephalochordate, amphioxus (Branchiostoma floridae) is thought to be the most basal living chordate. Gene products from amphioxus can be studied to gain insights into how the vertebrate orthologues of amphioxus genes evolved throughout chordate evolution. The present study set out to investigate and characterize biogenic amine GPCRs from amphioxus. To this end, the amino acid sequence of an amphioxus dopamine-like receptor, AmphiD1/β, was used to screen the B. floridae trace sequence database. Trace sequence files displaying similarity to AmphiD1/β were assembled into genes, and eleven novel aminergic receptor sequences were identified. AmphiD1/β and these were subject to a comprehensive bioinformatical analysis. From this analysis it was evident that amphioxus possesses receptors orthologous to the vertebrate dopamine receptors, α-adrenoreceptors, and histamine receptors, as well as receptors orthologous to the invertebrate dopamine receptors and octopamine receptors. PCR based techniques were then used to amplify the novel receptors from amphioxus cDNA libraries. To classify the novel receptors into specific biogenic amine receptor subtypes, detailed pharmacological analysis is required. Therefore, the present study also set out to pharmacologically characterize both AmphiD1/β and one of the novel receptors, AmphiAmR1. Both receptors were found to display dopamine D1-like receptor pharmacology, although the two receptors differed in respect to ligand specificity, second messenger signalling capabilities and levels of agonist-independent activity. Therefore, work undertaken in the present study clearly shows that amphioxus possesses at least two distinct dopamine D1-like receptors.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available