Characterization of the effects of four HTR3B polymorphisms an human 5-HT3AB receptor expression and signalling. / Krzywkowski, Karen; Davies, Paul A.; Irving, Andrew J.; Brauner-Osborne, Hans; Jensen, Anders A.
In: Pharmacogenetics and Genomics, Vol. 18, No. 12, 12.2008, p. 1027-1040.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Characterization of the effects of four HTR3B polymorphisms an human 5-HT3AB receptor expression and signalling
A1 - Krzywkowski,Karen
A1 - Davies,Paul A.
A1 - Irving,Andrew J.
A1 - Brauner-Osborne,Hans
A1 - Jensen,Anders A.
AU - Krzywkowski,Karen
AU - Davies,Paul A.
AU - Irving,Andrew J.
AU - Brauner-Osborne,Hans
AU - Jensen,Anders A.
PY - 2008/12
Y1 - 2008/12
N2 - <p>Background 5-Hydroxytryptamine 3 (5-HT3) receptors mediate the fast excitatory neurotransmission of serotonin. In this study, we have characterized the effects of four naturally occurring, nonsynonymous variants of the human 5-HT3B subunit on expression and signalling properties of heteromeric 5-HT3AB receptors.</p><p>Methods and results 5-HT3AB receptor signalling was studied in a fluorescence-based cell membrane potential assay and by electrophysiology. Expression levels of cotransfected epitope-tagged 5-HT3A and 5-HT3B subunits were determined using enzyme-linked immunosorbent assay and immunocytochemistry. In cells coexpressing 5-HT3A and 5-HT3B(I143T) subunits, cell surface expression levels of 5-HT3B in particular, and also 5-HT3A were markedly reduced compared with those of wild-type (WT) 5-HT3AB receptor-expressing cells. Electrophysiological recordings on cells coexpressing 5-HT3A and 5-HT3B(I143T) indicated cell surface expression of 5-HT3AB(1143T) receptors with macroscopic current kinetics similar to those of WT 5-HT3AB receptors but with 3-fold lower current densities. In the membrane potential assay, 5-HT3AB(I143T)-transfected cells exhibited signalling properties intermediate to those of WT 5-HT3AB and 5-HT3A receptors. Cotransfection of 5-HT3A, 5-HT3AB(I143T) and WT 5-HT3AB subunit cDNAs did not increase cell surface expression of the variant subunit nor did it restore WT 5-HT3AB receptor signalling completely in the membrane potential assay. In contrast to 5-HT3B(I143T), the 5-HT3B variants S1 56R, V183I and A223T did not give rise to significant changes in 5-HT3AB receptor expression or signalling properties.</p><p>Conclusion 5-HT3B(I143T)-containing 5-HT3AB receptors display significantly reduced cell surface expression and different signalling properties compared with WT 5-HT3AB receptors. In contrast, three other 5-HT3B variants, S156R, V183I and A223T, do not appear to alter 5-HT3AB receptor expression or signalling. Pharmacogenetics and Genomics 18:1027-1040 (C) 2008 Wolters Kluwer Health | Lippincott Williams & Wilkins.</p>
AB - <p>Background 5-Hydroxytryptamine 3 (5-HT3) receptors mediate the fast excitatory neurotransmission of serotonin. In this study, we have characterized the effects of four naturally occurring, nonsynonymous variants of the human 5-HT3B subunit on expression and signalling properties of heteromeric 5-HT3AB receptors.</p><p>Methods and results 5-HT3AB receptor signalling was studied in a fluorescence-based cell membrane potential assay and by electrophysiology. Expression levels of cotransfected epitope-tagged 5-HT3A and 5-HT3B subunits were determined using enzyme-linked immunosorbent assay and immunocytochemistry. In cells coexpressing 5-HT3A and 5-HT3B(I143T) subunits, cell surface expression levels of 5-HT3B in particular, and also 5-HT3A were markedly reduced compared with those of wild-type (WT) 5-HT3AB receptor-expressing cells. Electrophysiological recordings on cells coexpressing 5-HT3A and 5-HT3B(I143T) indicated cell surface expression of 5-HT3AB(1143T) receptors with macroscopic current kinetics similar to those of WT 5-HT3AB receptors but with 3-fold lower current densities. In the membrane potential assay, 5-HT3AB(I143T)-transfected cells exhibited signalling properties intermediate to those of WT 5-HT3AB and 5-HT3A receptors. Cotransfection of 5-HT3A, 5-HT3AB(I143T) and WT 5-HT3AB subunit cDNAs did not increase cell surface expression of the variant subunit nor did it restore WT 5-HT3AB receptor signalling completely in the membrane potential assay. In contrast to 5-HT3B(I143T), the 5-HT3B variants S1 56R, V183I and A223T did not give rise to significant changes in 5-HT3AB receptor expression or signalling properties.</p><p>Conclusion 5-HT3B(I143T)-containing 5-HT3AB receptors display significantly reduced cell surface expression and different signalling properties compared with WT 5-HT3AB receptors. In contrast, three other 5-HT3B variants, S156R, V183I and A223T, do not appear to alter 5-HT3AB receptor expression or signalling. Pharmacogenetics and Genomics 18:1027-1040 (C) 2008 Wolters Kluwer Health | Lippincott Williams & Wilkins.</p>
KW - 5-HT3A
KW - 5-HT3AB
KW - 5-HT3B
KW - expression
KW - function
KW - ligand-gated ion channel
KW - polymorphism
KW - serotonin
KW - single-nucleotide polymorphism
KW - variant
KW - NICOTINIC ACETYLCHOLINE-RECEPTORS
KW - SEROTONIN-RECEPTOR
KW - MAJOR DEPRESSION
KW - HUMAN GENOME
KW - SUBUNIT
KW - GENE
KW - 5-HYDROXYTRYPTAMINE
KW - CHANNEL
KW - HETEROGENEITY
KW - MUTATIONS
U2 - 10.1097/FPC.0b013e328310f950
DO - 10.1097/FPC.0b013e328310f950
M1 - Article
JO - Pharmacogenetics and Genomics
JF - Pharmacogenetics and Genomics
SN - 1744-6872
IS - 12
VL - 18
SP - 1027
EP - 1040
ER -