title: Forskolin modulation of desensitization at GABA(A) and glycine receptors is not mediated by cAMP-dependent protein kinase in isolated carp amacrine-like cells.
abstract: The effects of forskolin on gamma-aminobutyric acid type-A (GABA(A)) and glycine receptors in amacrine-like cells of carp (Carassius auratus) retina were studied using patch-clamp techniques. Application of 50 microM forskolin markedly accelerated the desensitization of whole-cell responses induced by 100 microM GABA or glycine without changing the peak amplitude of the response. Both 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP) and 3-isobutyl-1 -methylxanthine (IBMX) (500 microM) failed to accelerate the desensitization of these two receptors. Protein kinase A (PKA) inhibitors, N- [2-[(p-bromocinnamyl)amino]ethyl)-5-isoquinolinesulfonamide dihydrochloride (H-89) and N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride (H-8), could not block these effects of forskolin. An inactive analogue of forskolin, 1,9-dideoxyforskolin (DFSK), accelerated the desensitization effectively. These results suggest that forskolin's effects are not mediated by activation of the PKA pathway. Moreover, similar results were obtained using excised outside-out patches of these cells, suggesting that forskolin may act on an extracellular site(s). The neurosteroids 5alpha-pregnane-3alpha,21 -diol-20-one (THDOC) and 5-pregnen-31 -ol-20-one sulfate sodium (PS), structural analogues of forskolin, accelerated the desensitization of these receptors without changing the peak amplitudes, thus mimicking forskolin's effects. Furthermore, PS interacted with forskolin on these receptors so as to slow down the responses. These results raise the possibility that forskolin acts directly on an extracellular site(s) of the GABA(A) and glycine receptors, shared with neurosteroids, in carp amacrine-like cells.
authors.affiliation: Institute of Neurobiology, Fudan University and Shanghai Institute of Physiology, Chinese Academy of Sciences.
authors.forename: P
authors.initials: P
authors.lastname: Li
authors.forename: X L
authors.initials: XL
authors.lastname: Yang
mesh_terms: D000818:Animals; D005576:Colforsin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005998:Glycine / Q000494:pharmacology; D006054:Goldfish; D007546:Isoquinolines / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D011963:Receptors, GABA-A / Q000378:metabolism*; D018009:Receptors, Glycine / Q000378:metabolism*; D012160:Retina / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D013449:Sulfonamides; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D007546:Isoquinolines; D011963:Receptors, GABA-A; D018009:Receptors, Glycine; D013449:Sulfonamides; D005576:Colforsin; D005680:gamma-Aminobutyric Acid; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D005998:Glycine