Use this URL to cite or link to this record in EThOS:
Title: An investigation into the regulatory mechanisms associated with the recombinant human 5-hydroxytryptamine_1A (5-HT_1A) receptor
Author: Ruddell, Richard George
ISNI:       0000 0001 3539 3017
Awarding Body: University of Southampton
Current Institution: University of Southampton
Date of Award: 2001
Availability of Full Text:
Access from EThOS:
Full text unavailable from EThOS. Please try the link below.
Access from Institution:
Classically, acute agonist challenge results in a concomitant loss of G protein-coupled receptor signalling function and high affinity membrane ligand binding sites. Expression of the human 5-HT1A receptor in CHO-K1 cells allowed the study of a homogeneous receptor population expressed at high density. Exposure of CHO-K1 cells expressing the cloned 5-HT1A receptor to 1μM- 8-hydroxyl-2-(di-n-propylamino)-tetralin (8-hydroxy-DPAT) resulted in receptor internalisation (20% of receptors after 20 minutes) to an undefined intracellular compartment. Receptor internalisation was found to be sensitive to, hypertonic sucrose (0.7-0.8M) and Concanavalin A (0.5-1mg/ml), both of which are inhibitors of clathrin-mediated endocytosis. Removal of 8-hydroxy-DPAT from the incubation medium allowed 5-HT1A receptors to recycle back to the plasma membrane, this process occurred in the presence of the protein synthesis inhibitor cycloheximide and in a time dependent fashion (40-50 minutes). Mutagenesis of the putative PKC phosphorylation motifs, KRT149APR and effect upon receptor binding characteristics however, complete abolition of receptor redistribution in response to acute agonist challenge was observed. In a similar manner conservative and radical mutation of the internalisation motif NPVIY (Y400F and Y400A respectively) had no effect on receptor binding characteristics but they did abolish receptor internalisation in response to short term agonist incubation.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: Internalisation; Transcription; Phorphorylation