{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T01:21:34Z","timestamp":1791336094249,"version":"4.3.1"},"reference-count":36,"publisher":"National Academy of Sciences","issue":"5","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2001,2,27]]},"abstract":"<jats:p>\n                    To investigate the dynamics of guanosine 3\u2032,5\u2032-cyclic monophosphate\n (cGMP) in single living cells, we constructed genetically encoded,\n fluorescent cGMP indicators by bracketing cGMP-dependent protein kinase\n (cGPK), minus residues 1\u201377, between cyan and yellow mutants of green\n fluorescent protein. cGMP decreased fluorescence resonance energy\n transfer (FRET) and increased the ratio of cyan to yellow emissions by\n up to 1.5-fold with apparent dissociation constants of \u22482 \u03bcM and\n &gt;100:1 selectivity for cGMP over cAMP. To eliminate constitutive\n kinase activity, Thr\n                    <jats:sup>516<\/jats:sup>\n                    of cGPK was mutated to Ala.\n Emission ratio imaging of the indicators transfected into rat fetal\n lung fibroblast (RFL)-6 showed cGMP transients resulting from\n activation of soluble and particulate guanylyl cyclase, respectively,\n by nitric oxide (NO) and C-type natriuretic peptide (CNP). Whereas all\n naive cells tested responded to CNP, only 68% responded to NO. Both\n sets of signals showed large and variable (0.5\u20134 min) latencies. The\n phosphodiesterase (PDE) inhibitor 3-isobutyl-1-methylxanthine (IBMX)\n did not elevate cGMP on its own but consistently amplified responses to\n NO or CNP, suggesting that basal activity of guanylate cyclase is very\n low and emphasizing the importance of PDEs in cGMP recycling. A\n fraction of RFL cells showed slowly propagating tides of cGMP spreading\n across the cell in response to delocalized application of NO.\n Biolistically transfected Purkinje neurons showed cGMP responses to\n parallel fiber activity and NO donors, confirming that single-cell\n increases in cGMP occur under conditions appropriate to cause synaptic\n plasticity.\n                  <\/jats:p>","DOI":"10.1073\/pnas.051631298","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:36:44Z","timestamp":1027679804000},"page":"2437-2442","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":227,"title":["Spatiotemporal dynamics of guanosine 3\u2032,5\u2032-cyclic monophosphate revealed by a genetically encoded, fluorescent indicator"],"prefix":"10.1073","volume":"98","author":[{"given":"Akira","family":"Honda","sequence":"first","affiliation":[{"name":"Departments of Pharmacology and Molecular Physiology and\r Biophysics, University of Vermont, College of Medicine, Burlington, VT\r 05405-0068; and Department of Pharmacology and\r Howard Hughes Medical Institute, University of\r California at San Diego, La Jolla, CA 92093-0647"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen R.","family":"Adams","sequence":"additional","affiliation":[{"name":"Departments of Pharmacology and Molecular Physiology and\r Biophysics, University of Vermont, College of Medicine, Burlington, VT\r 05405-0068; and Department of Pharmacology and\r Howard Hughes Medical Institute, University of\r California at San Diego, La Jolla, CA 92093-0647"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolyn L.","family":"Sawyer","sequence":"additional","affiliation":[{"name":"Departments of Pharmacology and Molecular Physiology and\r Biophysics, University of Vermont, College of Medicine, Burlington, VT\r 05405-0068; and Department of Pharmacology and\r Howard Hughes Medical Institute, University of\r California at San Diego, La Jolla, CA 92093-0647"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Varda","family":"Lev-Ram","sequence":"additional","affiliation":[{"name":"Departments of Pharmacology and Molecular Physiology and\r Biophysics, University of Vermont, College of Medicine, Burlington, VT\r 05405-0068; and Department of Pharmacology and\r Howard Hughes Medical Institute, University of\r California at San Diego, La Jolla, CA 92093-0647"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roger Y.","family":"Tsien","sequence":"additional","affiliation":[{"name":"Departments of Pharmacology and Molecular Physiology and\r Biophysics, University of Vermont, College of Medicine, Burlington, VT\r 05405-0068; and Department of Pharmacology and\r Howard Hughes Medical Institute, University of\r California at San Diego, La Jolla, CA 92093-0647"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang R. G.","family":"Dostmann","sequence":"additional","affiliation":[{"name":"Departments of Pharmacology and Molecular Physiology and\r Biophysics, University of Vermont, College of Medicine, Burlington, VT\r 05405-0068; and Department of Pharmacology and\r Howard Hughes Medical Institute, University of\r California at San Diego, La Jolla, CA 92093-0647"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2001,2,27]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0033670"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0033671"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1054-3589(08)61094-7"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0033672"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0033673"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0033668"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0033669"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0959-4388(00)00101-X"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/42264"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(00)27297-2"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/71345"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/349694a0"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac0006167"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.22.11996"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(89)81453-X"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(95)90171-X"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)81196-4"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(20)79660-2"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.286.5444.1583"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4838(84)90025-6"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.50.31929"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1987.tb13395.x"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(96)01242-2"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.16.10522"},{"key":"e_1_3_3_25_2","volume-title":"Habilitationsschrift","author":"Dostmann W R G","year":"1996","unstructured":"W R G Dostmann Habilitationsschrift (Technische Universit\u00e4t, M\u00fcnchen, 1996)."},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00040a029"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1991.tb16509.x"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1992.tb17209.x"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.77.9.5537"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm00115a013"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(95)90044-6"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80340-2"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80099-9"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4889(86)90040-6"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.3.1188"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80604-2"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.051631298","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T15:03:07Z","timestamp":1649775787000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.051631298"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,2,27]]},"references-count":36,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2001,2,27]]}},"alternative-id":["10.1073\/pnas.051631298"],"URL":"https:\/\/doi.org\/10.1073\/pnas.051631298","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,2,27]]},"assertion":[{"value":"2000-12-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2001-02-27","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}