title: AKAP79/150 signal complexes in G-protein modulation of neuronal ion channels.
abstract: Voltage-gated M-type (KCNQ) K+ channels play critical roles in regulation of neuronal excitability. Previous work showed A-kinase-anchoring protein (AKAP)79/150-mediated protein kinase C (PKC) phosphorylation of M channels to be involved in M current (I(M)) suppression by muscarinic M1, but not bradykinin B2, receptors. In this study, we first explored whether purinergic and angiotensin suppression of I(M) in superior cervical ganglion (SCG) sympathetic neurons involves AKAP79/150. Transfection into rat SCG neurons of ΔA-AKAP79, which lacks the A domain necessary for PKC binding, or the absence of AKAP150 in AKAP150(-/-) mice, did not affect I(M) suppression by purinergic agonist or by bradykinin, but reduced I(M) suppression by muscarinic agonist and angiotensin II. Transfection of AKAP79, but not ΔA-AKAP79 or AKAP15, rescued suppression of I(M) by muscarinic receptors in AKAP150(-/-) neurons. We also tested association of AKAP79 with M(1), B(2), P2Y(6), and AT(1) receptors, and KCNQ2 and KCNQ3 channels, via Förster resonance energy transfer (FRET) on Chinese hamster ovary cells under total internal refection fluorescence microscopy, which revealed substantial FRET between AKAP79 and M1 or AT1 receptors, and with the channels, but only weak FRET with P2Y(6) or B2 receptors. The involvement of AKAP79/150 in G(q/11)-coupled muscarinic regulation of N- and L-type Ca2+) channels and by cAMP/protein kinase A was also studied. We found AKAP79/150 to not play a role in the former, but to be necessary for forskolin-induced upregulation of L-current. Thus, AKAP79/150 action correlates with the PIP(2) (phosphatidylinositol 4,5-bisphosphate)-depletion mode of I(M) suppression, but does not generalize to G(q/11)-mediated inhibition of N- or L-type Ca2+ channels.
authors.affiliation: Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229, USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Manjot
authors.initials: M
authors.lastname: Bal
authors.forename: Sonya
authors.initials: S
authors.lastname: Bierbower
authors.forename: Oleg
authors.initials: O
authors.lastname: Zaika
authors.forename: Mark S
authors.initials: MS
authors.lastname: Shapiro
mesh_terms: D054758:A Kinase Anchor Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D019204:GTP-Binding Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D015640:Ion Channel Gating / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D017783:Superior Cervical Ganglion / Q000166:cytology / Q000378:metabolism
chemical_list: D054758:A Kinase Anchor Proteins; D019204:GTP-Binding Proteins