ArticleProceedings of the National Academy of Sciences of the United States of America2001
Spatiotemporal dynamics of guanosine 3',5'-cyclic monophosphate revealed by a genetically encoded, fluorescent indicator.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 72 papers.
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Who cites it
72 citing papers in PubMed.
- Molecular Spies in Action: Genetically Encoded Fluorescent Biosensors Light up Cellular Signals.Chemical reviews · 2024Review
- Hyperspectral imaging and dynamic region of interest tracking approaches to quantify localized cAMP signals.Biochemical Society transactions · 2024Review
- A Homogeneous Bioluminescent System to Monitor Cyclic Guanosine Monophosphate.ACS pharmacology & translational science · 2023Article
- Transgenic mice encoding modern imaging probes: Properties and applications.Cell reports · 2022Review
- CNP regulates cardiac contractility and increases cGMP near both SERCA and TnI: difference from BNP visualized by targeted cGMP biosensors.Cardiovascular research · 2022Article
- A Real-Time, Plate-Based BRET Assay for Detection of cGMP in Primary Cells.International journal of molecular sciences · 2022Article
- Recent progress in developing fluorescent probes for imaging cell metabolites.Biomedical materials (Bristol, England) · 2021Review
- Spatiotemporal Measurement of Osmotic Pressures by FRET Imaging.Angewandte Chemie (International ed. in English) · 2021Article
- Nitric oxide resets kisspeptin-excited GnRH neurons via PIP2 replenishment.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- CUTie2: The Attack of the Cyclic Nucleotide Sensor Clones.Frontiers in molecular biosciences · 2021Article
- Genetically encoded FRET-based optical sensor for HgJournal of industrial microbiology & biotechnology · 2019Article
- Detection and quantification of nitric oxide-derived oxidants in biological systems.The Journal of biological chemistry · 2019Review
- Red Fluorescent Genetically Encoded Voltage Indicators with Millisecond Responsiveness.Sensors (Basel, Switzerland) · 2019Article
- FRET-based cyclic GMP biosensors measure low cGMP concentrations in cardiomyocytes and neurons.Communications biology · 2019Article
- Genetically Encoded Fluorescent Biosensors Illuminate the Spatiotemporal Regulation of Signaling Networks.Chemical reviews · 2018Review
- cGMP Signaling in the Cardiovascular System-The Role of Compartmentation and Its Live Cell Imaging.International journal of molecular sciences · 2018Review
- Imaging of PDE2- and PDE3-Mediated cGMP-to-cAMP Cross-Talk in Cardiomyocytes.Journal of cardiovascular development and disease · 2018Review
- Integrated Strategies to Gain a Systems-Level View of Dynamic Signaling Networks.Methods in enzymology · 2017Article
- Modulation of Compartmentalised Cyclic Nucleotide Signalling via Local Inhibition of Phosphodiesterase Activity.International journal of molecular sciences · 2016Review
- Design and development of genetically encoded fluorescent sensors to monitor intracellular chemical and physical parameters.Biophysical reviews · 2016Review
12 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors.
Funding
Abstract
To investigate the dynamics of guanosine 3',5'-cyclic monophosphate (cGMP) in single living cells, we constructed genetically encoded, fluorescent cGMP indicators by bracketing cGMP-dependent protein kinase (cGPK), minus residues 1-77, between cyan and yellow mutants of green fluorescent protein. cGMP decreased fluorescence resonance energy transfer (FRET) and increased the ratio of cyan to yellow emissions by up to 1.5-fold with apparent dissociation constants of approximately 2 microM and >100:1 selectivity for cGMP over cAMP. To eliminate constitutive kinase activity, Thr(516) of cGPK was mutated to Ala. Emission ratio imaging of the indicators transfected into rat fetal lung fibroblast (RFL)-6 showed cGMP transients resulting from activation of soluble and particulate guanylyl cyclase, respectively, by nitric oxide (NO) and C-type natriuretic peptide (CNP). Whereas all naive cells tested responded to CNP, only 68% responded to NO. Both sets of signals showed large and variable (0.5-4 min) latencies. The phosphodiesterase (PDE) inhibitor 3-isobutyl-1-methylxanthine (IBMX) did not elevate cGMP on its own but consistently amplified responses to NO or CNP, suggesting that basal activity of guanylate cyclase is very low and emphasizing the importance of PDEs in cGMP recycling. A fraction of RFL cells showed slowly propagating tides of cGMP spreading across the cell in response to delocalized application of NO. Biolistically transfected Purkinje neurons showed cGMP responses to parallel fiber activity and NO donors, confirming that single-cell increases in cGMP occur under conditions appropriate to cause synaptic plasticity.
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