title: Transfection with 5-HT1A receptor gene and antisense directed against muscarinic M2 receptors reveal a mutual influence between Gi/o-coupled receptors in rat atrial myocytes.
abstract: A recently described reduction in sensitivity of G protein-activated inward-rectifying K(+) (GIRK) channels to stimulation of muscarinic M(2) receptors (M(2)AChR) in atrial myocytes overexpressing purinergic A(1) receptors (A(1)AdoR) was further investigated by heterologous expression of a 5-HT(1A) receptor (5-HT(1A)R) and by reducing the expression level of endogenous M(2)AChR receptors using antisense. In 5-HT(1A)R-expressing myocytes, in line with previous studies, sizable GIRK currents could be activated by 5-HT. In these cells, the mean current density and activation rate of M(2)AChR-activated current were significantly reduced, supporting the notion that signalling via this receptor is negatively regulated by other G protein-coupled receptors (GPCR) coupling to the same class (G(i/o)) of G proteins. To study if reducing M(2)AChR expression affects sensitivity of GIRK current to stimulation of A(1)AdoR, antisense oligodinucleotides (AsODN) against the M(2)AChR were used. Incubation of myocytes with M(2)AChR-specific AsODN resulted in a significant reduction in mean amplitude and activation rate of ACh-induced currents. This was paralleled by an increase in mean amplitude and activation rate of current activated by stimulation of A(1)AdoR. Plotting amplitudes of 5-HT- or Ado-induced currents from individual manipulated cells against the amplitude of ACh-induced current yielded a positive correlation between these data. Although difficult to interpret in mechanistic terms, this argues against a competition of receptors for a common pool of G(i/o). The mutual interaction between G(i/o)-coupled receptors depends on manipulation of the expression level, since long-term desensitization or down regulation of M(2)AChR by treatment with carbachol did not affect sensitivity of GIRK current to A(1)AdoR stimulation, despite a substantial reduction in amplitude and activation rate of M(2)AChR-activated currents. These data suggest a novel crosstalk between parallel receptors converging on the same class of G proteins.
authors.affiliation: Department of Physiology, Ruhr-University Bochum, D-4480, Bochum, Germany.
authors.forename: Leif I
authors.initials: LI
authors.lastname: Bösche
authors.forename: Kirsten
authors.initials: K
authors.lastname: Bender
authors.forename: Marie Cécile
authors.initials: MC
authors.lastname: Wellner-Kienitz
authors.forename: Lutz
authors.initials: L
authors.lastname: Pott
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000235:genetics / Q000502:physiology; D005260:Female; D008297:Male; D016376:Oligonucleotides, Antisense; D051381:Rats; D043585:Receptor, Muscarinic M2; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000235:genetics* / Q000378:metabolism; D011985:Receptors, Serotonin / Q000235:genetics* / Q000378:metabolism; D044263:Receptors, Serotonin, 5-HT1; D014162:Transfection
chemical_list: D016376:Oligonucleotides, Antisense; D043585:Receptor, Muscarinic M2; D011956:Receptors, Cell Surface; D011976:Receptors, Muscarinic; D011985:Receptors, Serotonin; D044263:Receptors, Serotonin, 5-HT1