Fluorescent biosensors illuminate the spatial regulation of cell signaling across scales

As cell signaling research has advanced, it has become clearer that signal transduction has complex spatiotemporal regulation that goes beyond foundational linear transduction models. Several technologies have enabled these discoveries, including fluorescent biosensors designed to report live biochemical signaling events. As genetically encoded and live-cell compatible tools, fluorescent biosensors are well suited to address diverse cell signaling questions across different spatial scales of regulation. In this review, methods of examining spatial signaling regulation and the design of fluorescent biosensors are introduced. Then, recent biosensor developments that illuminate the importance of spatial regulation in cell signaling are highlighted at several scales, including membranes and organelles, molecular assemblies, and cell/tissue heterogeneity. In closing, perspectives on how fluorescent biosensors will continue enhancing cell signaling research are discussed.

[1]  Jin Zhang,et al.  Fluorescent biosensor imaging meets deterministic mathematical modelling: quantitative investigation of signalling compartmentalization , 2023, The Journal of physiology.

[2]  S. Cappell,et al.  Loss of CDK4/6 activity in S/G2 phase leads to cell cycle reversal , 2023, Nature.

[3]  R. Sunahara,et al.  Non-canonical β-adrenergic activation of ERK at endosomes , 2022, Nature.

[4]  Michelle C. Mendoza,et al.  ERK signaling for cell migration and invasion , 2022, Frontiers in Molecular Biosciences.

[5]  X. Trepat,et al.  A feedback loop between lamellipodial extension and HGF-ERK signaling specifies leader cells during collective cell migration. , 2022, Developmental cell.

[6]  L. Pyle,et al.  Functional architecture of pancreatic islets identifies a population of first responder cells that drive the first-phase calcium response , 2022, PLoS biology.

[7]  A. Lita,et al.  Advances in measuring cancer cell metabolism with subcellular resolution , 2022, Nature Methods.

[8]  R. Shaw,et al.  Spatial regulation of AMPK signaling revealed by a sensitive kinase activity reporter , 2022, Nature Communications.

[9]  K. Hiller,et al.  Complexity of subcellular metabolism: strategies for compartment-specific profiling. , 2022, Current opinion in biotechnology.

[10]  J. Brickman,et al.  Transcriptional heterogeneity and cell cycle regulation as central determinants of Primitive Endoderm priming , 2022, bioRxiv.

[11]  M. Zaccolo,et al.  Receptor-associated independent cAMP nanodomains mediate spatiotemporal specificity of GPCR signaling , 2022, Cell.

[12]  K. Guan,et al.  Rheb regulates nuclear mTORC1 activity independent of farnesylation. , 2022, Cell chemical biology.

[13]  Jin Zhang,et al.  Liquid–liquid phase separation drives cellular function and dysfunction in cancer , 2022, Nature Reviews Cancer.

[14]  O. Bergmann,et al.  FUCCI-Based Live Imaging Platform Reveals Cell Cycle Dynamics and Identifies Pro-proliferative Compounds in Human iPSC-Derived Cardiomyocytes , 2021, bioRxiv.

[15]  Yanghao Zhong,et al.  A Highly Sensitive Fluorescent Akt Biosensor Reveals Lysosome-Selective Regulation of Lipid Second Messengers and Kinase Activity , 2021, ACS central science.

[16]  P. Iglesias,et al.  Deciphering cell signaling networks with massively multiplexed biosensor barcoding , 2021, Cell.

[17]  J. Teruya-Feldstein,et al.  Integrative RNA-omics discovers GNAS alternative splicing as a phenotypic driver of splicing factor-mutant neoplasms. , 2021, Cancer discovery.

[18]  R. Shaw,et al.  AMPK: restoring metabolic homeostasis over space and time. , 2021, Molecular cell.

[19]  Jin Zhang,et al.  DrFLINC Contextualizes Super-resolution Activity Imaging. , 2021, Journal of the American Chemical Society.

[20]  K. Chung,et al.  Scaffolding mechanism of arrestin-2 in the cRaf/MEK1/ERK signaling cascade , 2021, Proceedings of the National Academy of Sciences.

[21]  Govind Menon,et al.  Spatial localisation meets biomolecular networks , 2021, Nature Communications.

[22]  Clint L. Miller,et al.  Mitochondria-localized AMPK responds to local energetics and contributes to exercise and energetic stress-induced mitophagy , 2021, Proceedings of the National Academy of Sciences.

[23]  R. Brooks.  Cell Cycle Commitment and the Origins of Cell Cycle Variability , 2021, Frontiers in Cell and Developmental Biology.

[24]  C. Parent,et al.  The principles of directed cell migration , 2021, Nature Reviews Molecular Cell Biology.

[25]  Sohum Mehta,et al.  FluoSTEPs: Fluorescent biosensors for monitoring compartmentalized signaling within endogenous microdomains , 2021, Science Advances.

[26]  F. Matschinsky,et al.  Metabolic Homeostasis in Life as We Know It: Its Origin and Thermodynamic Basis , 2021, Frontiers in Physiology.

[27]  R. Campbell,et al.  Structure- and mechanism-guided design of single fluorescent protein-based biosensors , 2021, Nature Chemical Biology.

[28]  J. Albeck,et al.  Transient phases of OXPHOS inhibitor resistance reveal underlying metabolic heterogeneity in single cells. , 2021, Cell metabolism.

[29]  M. Caron,et al.  Noncanonical scaffolding of Gαi and β-arrestin by G protein–coupled receptors , 2021, Science.

[30]  A. Ting,et al.  Deciphering molecular interactions by proximity labeling , 2021, Nature Methods.

[31]  Suwen Zhao,et al.  G protein-coupled receptors: structure- and function-based drug discovery , 2021, Signal Transduction and Targeted Therapy.

[32]  M. Zaccolo,et al.  Subcellular Organization of the cAMP Signaling Pathway , 2020, Pharmacological Reviews.

[33]  A. Newton,et al.  Location-specific inhibition of Akt reveals regulation of mTORC1 activity in the nucleus , 2020, Nature Communications.

[34]  Katarzyna P. Adamala,et al.  Spatial Multiplexing of Fluorescent Reporters for Imaging Signaling Network Dynamics , 2020, Cell.

[35]  M. Holz,et al.  Unraveling the Multifaceted Nature of the Nuclear Function of mTOR. , 2020, Biochimica et biophysica acta. Molecular cell research.

[36]  B. Giepmans,et al.  Nanobody-Based Probes for Subcellular Protein Identification and Visualization , 2020, Frontiers in Cellular Neuroscience.

[37]  John D. Scott,et al.  AKAP Signaling Islands: Venues for Precision Pharmacology. , 2020, Trends in pharmacological sciences.

[38]  Claire S. Simon,et al.  Live Visualization of ERK Activity in the Mouse Blastocyst Reveals Lineage-Specific Signaling Dynamics. , 2020, Developmental cell.

[39]  W. Timp,et al.  Cell-Cycle-Dependent ERK Signaling Dynamics Direct Fate Specification in the Mammalian Preimplantation Embryo. , 2020, Developmental cell.

[40]  U. Zabel,et al.  Optical Mapping of cAMP Signaling at the Nanometer Scale , 2020, Cell.

[41]  Lucas M. Stolerman,et al.  Phase Separation of a PKA Regulatory Subunit Controls cAMP Compartmentation and Oncogenic Signaling , 2020, Cell.

[42]  Paul Martin,et al.  Live-imaging of endothelial Erk activity reveals dynamic and sequential signalling events during regenerative angiogenesis , 2020, bioRxiv.

[43]  Andrew R. Cohen,et al.  Collective ERK/Akt activity waves orchestrate epithelial homeostasis by driving apoptosis-induced survival , 2020, bioRxiv.

[44]  Brian L Ross,et al.  Spatially compartmentalized phase regulation of a Ca2+-cAMP-PKA oscillatory circuit , 2020, bioRxiv.

[45]  X. Trepat,et al.  ERK-mediated mechanochemical waves direct collective cell polarization , 2019, bioRxiv.

[46]  Jeremiah D. Keyes,et al.  Signaling diversity enabled by Rap1-regulated plasma membrane ERK with distinct temporal dynamics , 2019, bioRxiv.

[47]  J. Fletcher,et al.  Visualizing Engrafted Human Cancer and Therapy Responses in Immunodeficient Zebrafish , 2019, Cell.

[48]  J. Bonifacino,et al.  Lysosome Positioning Influences mTORC2 and AKT Signaling. , 2019, Molecular cell.

[49]  M. Caron,et al.  Noncanonical scaffolding of Gαi and β-arrestin by G protein-coupled receptors , 2019, bioRxiv.

[50]  G. Fairn,et al.  Akt-ing Up Just About Everywhere: Compartment-Specific Akt Activation and Function in Receptor Tyrosine Kinase Signaling , 2019, Front. Cell Dev. Biol..

[51]  A. Bode,et al.  AKT as a Therapeutic Target for Cancer. , 2019, Cancer research.

[52]  D. Carling,et al.  AMP-activated protein kinase: the current landscape for drug development , 2019, Nature Reviews Drug Discovery.

[53]  R. Satija,et al.  Integrative single-cell analysis , 2019, Nature Reviews Genetics.

[54]  M. Sauer,et al.  Super-resolution microscopy demystified , 2019, Nature Cell Biology.

[55]  Gerald R. V. Hammond,et al.  A high-avidity biosensor reveals plasma membrane PI(3,4)P2 is predominantly a class I PI3K signaling product , 2018, The Journal of cell biology.

[56]  P. Meikle,et al.  Inhibition of Adenosine Monophosphate–Activated Protein Kinase–3‐Hydroxy‐3‐Methylglutaryl Coenzyme A Reductase Signaling Leads to Hypercholesterolemia and Promotes Hepatic Steatosis and Insulin Resistance , 2018, Hepatology communications.

[57]  Paul J. Hoffman,et al.  Comprehensive Integration of Single-Cell Data , 2018, Cell.

[58]  R. Lefkowitz,et al.  Manifold roles of β-arrestins in GPCR signaling elucidated with siRNA and CRISPR/Cas9 , 2018, Science Signaling.

[59]  Sohum Mehta,et al.  Single-fluorophore Biosensors for Sensitive and Multiplexed Detection of Signaling Activities , 2018, Nature Cell Biology.

[60]  N. Perrimon,et al.  Efficient proximity labeling in living cells and organisms with TurboID , 2018, Nature Biotechnology.

[61]  Rosa A. Uribe,et al.  Tracking neural crest cell cycle progression in vivo , 2018, Genesis.

[62]  Benjamin M. Gyori,et al.  Encoding Growth Factor Identity in the Temporal Dynamics of FOXO3 under the Combinatorial Control of ERK and AKT Kinases. , 2018, Cell systems.

[63]  A. Brand,et al.  Cell cycle heterogeneity directs the timing of neural stem cell activation from quiescence , 2018, Science.

[64]  F. Pinaud,et al.  Characterization of Split Fluorescent Protein Variants and Quantitative Analyses of Their Self-Assembly Process , 2018, Scientific Reports.

[65]  I. Yampolsky,et al.  SypHer3s: a genetically encoded fluorescent ratiometric probe with enhanced brightness and an improved dynamic range. , 2018, Chemical communications.

[66]  J. Wess,et al.  Lack of beta-arrestin signaling in the absence of active G proteins , 2018, Nature Communications.

[67]  J. Skotheim,et al.  A Precise Cdk Activity Threshold Determines Passage through the Restriction Point. , 2018, Molecular cell.

[68]  Derek N. Macklin,et al.  Live-cell measurements of kinase activity in single cells using translocation reporters , 2017, Nature Protocols.

[69]  Matthew J Simpson,et al.  Mathematical models for cell migration with real-time cell cycle dynamics , 2017, bioRxiv.

[70]  Michiyuki Matsuda,et al.  A Highly Sensitive FRET Biosensor for AMPK Exhibits Heterogeneous AMPK Responses among Cells and Organs. , 2017, Cell reports.

[71]  Kazuhiro Aoki,et al.  Propagating Wave of ERK Activation Orients Collective Cell Migration. , 2017, Developmental cell.

[72]  R. Shaw,et al.  AMPK: guardian of metabolism and mitochondrial homeostasis , 2017, Nature Reviews Molecular Cell Biology.

[73]  G. Kroemer,et al.  Calcium signaling and cell cycle: Progression or death. , 2017, Cell calcium.

[74]  R. Sunahara,et al.  Genetic evidence that β-arrestins are dispensable for the initiation of β2-adrenergic receptor signaling to ERK , 2017, Science Signaling.

[75]  M. von Zastrow,et al.  Functional selectivity of GPCR-directed drug action through location bias , 2017, Nature chemical biology.

[76]  L. Schad,et al.  Experimental and mathematical analysis of cAMP nanodomains , 2017, PloS one.

[77]  G. Stephanopoulos,et al.  Akt regulation of glycolysis mediates bioenergetic stability in epithelial cells , 2017, bioRxiv.

[78]  M. Miura,et al.  Radiosensitivity of quiescent and proliferating cells grown as multicellular tumor spheroids , 2017, Cancer science.

[79]  Peter Dedecker,et al.  Genetically-Encoded Biosensors for Visualizing Live-cell Biochemical Activity at Superresolution , 2017, Nature Methods.

[80]  G. Ronnett,et al.  AKAP-mediated feedback control of cAMP gradients in developing hippocampal neurons , 2017, Nature chemical biology.

[81]  R. Strack,et al.  Expansion microscopy , 2016, Nature Methods.

[82]  Tudor I. Oprea,et al.  A comprehensive map of molecular drug targets , 2016, Nature Reviews Drug Discovery.

[83]  Michael Z. Lin,et al.  Fluorescent indicators for simultaneous reporting of all four cell cycle phases , 2016, Nature Methods.

[84]  W. Cui,et al.  Proliferation, survival and metabolism: the role of PI3K/AKT/mTOR signalling in pluripotency and cell fate determination , 2016, Development.

[85]  Michael Z. Lin,et al.  A Guide to Fluorescent Protein FRET Pairs , 2016, Sensors.

[86]  Daria M. Shcherbakova,et al.  Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging , 2016, Nature Communications.

[87]  Roger Y Tsien,et al.  A far-red fluorescent protein evolved from a cyanobacterial phycobiliprotein , 2016, Nature Methods.

[88]  Luke A. Gilbert,et al.  Versatile protein tagging in cells with split fluorescent protein , 2016, Nature Communications.

[89]  R. Chhipa,et al.  Evolving Lessons on the Complex Role of AMPK in Normal Physiology and Cancer. , 2016, Trends in pharmacological sciences.

[90]  Stephanie S. Lam,et al.  Spatially resolved proteomic mapping in living cells with the engineered peroxidase APEX2 , 2016, Nature Protocols.

[91]  Colleen E. Clancy,et al.  Mechanisms Restricting Diffusion of Intracellular cAMP , 2016, Scientific Reports.

[92]  Takeshi Imamura,et al.  Quantitative imaging with Fucci and mathematics to uncover temporal dynamics of cell cycle progression , 2016, Development, growth & differentiation.

[93]  Shigeo Okabe,et al.  Fluorescent ratiometric pH indicator SypHer2: Applications in neuroscience and regenerative biology. , 2015, Biochimica et biophysica acta.

[94]  Roy Wollman,et al.  Paracrine communication maximizes cellular response fidelity in wound signaling , 2015, eLife.

[95]  Michael Pargett,et al.  Receptor Level Mechanisms Are Required for Epidermal Growth Factor (EGF)-stimulated Extracellular Signal-regulated Kinase (ERK) Activity Pulses* , 2015, The Journal of Biological Chemistry.

[96]  A. Cuervo,et al.  Lysosomal mTORC2/PHLPP1/Akt Regulate Chaperone-Mediated Autophagy. , 2015, Molecular cell.

[97]  Aiman Alam-Nazki,et al.  Spatial Control of Biochemical Modification Cascades and Pathways. , 2015, Biophysical journal.

[98]  Igor Stagljar,et al.  Application guide for omics approaches to cell signaling. , 2015, Nature chemical biology.

[99]  Robert G. Endres,et al.  Accurate Encoding and Decoding by Single Cells: Amplitude Versus Frequency Modulation , 2015, PLoS Comput. Biol..

[100]  J. Blenis,et al.  ERK reinforces actin polymerization to power persistent edge protrusion during motility , 2015, Science Signaling.

[101]  H. Tokumitsu,et al.  Compartmentalized AMPK signaling illuminated by genetically encoded molecular sensors and actuators. , 2015, Cell reports.

[102]  Gaudenz Danuser,et al.  In vivo cell cycle profiling in xenograft tumors by quantitative intravital microscopy , 2015, Nature Methods.

[103]  Xin Zhou,et al.  Dynamic Visualization of mTORC1 Activity in Living Cells. , 2015, Cell reports.

[104]  Honda Naoki,et al.  Intercellular propagation of extracellular signal-regulated kinase activation revealed by in vivo imaging of mouse skin , 2015, eLife.

[105]  Edward S. Boyden,et al.  Expansion microscopy , 2015, Science.

[106]  A. Miyawaki,et al.  Fucci-guided purification of hematopoietic stem cells with high repopulating activity. , 2015, Biochemical and biophysical research communications.

[107]  Susan S. Taylor,et al.  Using Markov State Models to Develop a Mechanistic Understanding of Protein Kinase A Regulatory Subunit RIα Activation in Response to cAMP Binding* , 2014, The Journal of Biological Chemistry.

[108]  N. Haass,et al.  Real‐time cell cycle imaging during melanoma growth, invasion, and drug response , 2014, Pigment cell & melanoma research.

[109]  S. Kim,et al.  AMPKα2 translocates into the nucleus and interacts with hnRNP H: implications in metformin-mediated glucose uptake. , 2014, Cellular signalling.

[110]  A. Miyawaki,et al.  Time-lapse imaging of cell cycle dynamics during development in living cardiomyocyte. , 2014, Journal of molecular and cellular cardiology.

[111]  M. Bouvet,et al.  Spatial–temporal FUCCI imaging of each cell in a tumor demonstrates locational dependence of cell cycle dynamics and chemoresponsiveness , 2014, Cell cycle.

[112]  C. Porta,et al.  Targeting PI3K/Akt/mTOR Signaling in Cancer , 2014, Front. Oncol..

[113]  Jie H. S. Zhou,et al.  Stretched cell cycle model for proliferating lymphocytes , 2014, Proceedings of the National Academy of Sciences.

[114]  M. von Zastrow,et al.  GPCR signaling along the endocytic pathway. , 2014, Current opinion in cell biology.

[115]  Ludovic Vallier,et al.  The Cell-Cycle State of Stem Cells Determines Cell Fate Propensity , 2014, Cell.

[116]  V. Vacic,et al.  Detection of a Recurrent DNAJB1-PRKACA Chimeric Transcript in Fibrolamellar Hepatocellular Carcinoma , 2014, Science.

[117]  L. Weisman,et al.  Genetically encoded fluorescent probe to visualize intracellular phosphatidylinositol 3,5-bisphosphate localization and dynamics , 2013, Proceedings of the National Academy of Sciences.

[118]  John D. Scott,et al.  AKAP signaling complexes: pointing towards the next generation of therapeutic targets? , 2013, Trends in pharmacological sciences.

[119]  C. Betz,et al.  Where is mTOR and what is it doing there? , 2013, The Journal of cell biology.

[120]  Kazuhiro Aoki,et al.  Stochastic ERK activation induced by noise and cell-to-cell propagation regulates cell density-dependent proliferation. , 2013, Molecular cell.

[121]  Sabrina L. Spencer,et al.  The Proliferation-Quiescence Decision Is Controlled by a Bifurcation in CDK2 Activity at Mitotic Exit , 2013, Cell.

[122]  D. Hardie,et al.  AMP Is a True Physiological Regulator of AMP-Activated Protein Kinase by Both Allosteric Activation and Enhancing Net Phosphorylation , 2013, Cell metabolism.

[123]  S. Pauklin,et al.  The Cell-Cycle State of Stem Cells Determines Cell Fate Propensity , 2013, Cell.

[124]  John G. Albeck,et al.  Frequency-modulated pulses of ERK activity transmit quantitative proliferation signals. , 2013, Molecular cell.

[125]  J. Draper,et al.  Lengthened G1 phase indicates differentiation status in human embryonic stem cells. , 2013, Stem cells and development.

[126]  A. Martelli,et al.  The emerging multiple roles of nuclear Akt. , 2012, Biochimica et biophysica acta.

[127]  Michael Z. Lin,et al.  Improving FRET dynamic range with bright green and red fluorescent proteins , 2012, Nature Methods.

[128]  M. Zaccolo,et al.  PKA and PDE4D3 anchoring to AKAP9 provides distinct regulation of cAMP signals at the centrosome , 2012, The Journal of cell biology.

[129]  K. Dodge-Kafka,et al.  AKAPs: the architectural underpinnings of local cAMP signaling. , 2012, Journal of molecular and cellular cardiology.

[130]  Kazuhiro Aoki,et al.  Development of an optimized backbone of FRET biosensors for kinases and GTPases , 2011, Molecular biology of the cell.

[131]  Wolfgang Link,et al.  Protein localization in disease and therapy , 2011, Journal of Cell Science.

[132]  D. Dickson,et al.  Nuclear translocation of AMPK-α1 potentiates striatal neurodegeneration in Huntington’s disease , 2011, The Journal of cell biology.

[133]  K. Matlin.  Spatial expression of the genome: the signal hypothesis at forty , 2011, Nature Reviews Molecular Cell Biology.

[134]  M. West,et al.  Origin of bistability underlying mammalian cell cycle entry , 2011, Molecular systems biology.

[135]  Mengwei Zang,et al.  AMPK phosphorylates and inhibits SREBP activity to attenuate hepatic steatosis and atherosclerosis in diet-induced insulin-resistant mice. , 2011, Cell metabolism.

[136]  L. Cantley,et al.  A fluorescent reporter of AMPK activity and cellular energy stress. , 2011, Cell metabolism.

[137]  B. Viollet,et al.  AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1 , 2011, Nature Cell Biology.

[138]  S. Rasmussen,et al.  Structure of a nanobody-stabilized active state of the β2 adrenoceptor , 2010, Nature.

[139]  Y. Yarden,et al.  Roles for growth factors in cancer progression. , 2010, Physiology.

[140]  Hoyun Lee,et al.  The Akt isoforms are present at distinct subcellular locations. , 2010, American journal of physiology. Cell physiology.

[141]  T. Pawson,et al.  Cell Signaling in Space and Time: Where Proteins Come Together and When They’re Apart , 2009, Science.

[142]  P. R. ten Wolde,et al.  Spatio-temporal correlations can drastically change the response of a MAPK pathway , 2009, Proceedings of the National Academy of Sciences.

[143]  A. Miyawaki,et al.  Tracing the silhouette of individual cells in S/G2/M phases with fluorescence. , 2008, Chemistry & biology.

[144]  K. Svoboda,et al.  A genetically encoded fluorescent sensor of ERK activity , 2008, Proceedings of the National Academy of Sciences.

[145]  M. Mank,et al.  Genetically encoded calcium indicators. , 2008, Chemical reviews.

[146]  Tae J. Lee,et al.  A bistable Rb–E2F switch underlies the restriction point , 2008, Nature Cell Biology.

[147]  Atsushi Miyawaki,et al.  Visualizing Spatiotemporal Dynamics of Multicellular Cell-Cycle Progression , 2008, Cell.

[148]  Yuichiro Hori,et al.  [Crystal structure of the Aequorea victoria green fluorescent protein]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[149]  D. Carling,et al.  Investigating the mechanism for AMP activation of the AMP-activated protein kinase cascade. , 2007, The Biochemical journal.

[150]  Eytan Domany,et al.  A module of negative feedback regulators defines growth factor signaling , 2007, Nature Genetics.

[151]  Daniel A. Haber,et al.  Epidermal growth factor receptor mutations in lung cancer , 2007, Nature Reviews Cancer.

[152]  P. Bastiaens,et al.  Growth factor-induced MAPK network topology shapes Erk response determining PC-12 cell fate , 2007, Nature Cell Biology.

[153]  J. Gutkind,et al.  G-protein-coupled receptors and cancer , 2007, Nature Reviews Cancer.

[154]  E. Golemis,et al.  Interdependence of cell attachment and cell cycle signaling. , 2006, Current opinion in cell biology.

[155]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[156]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[157]  J. Zierath,et al.  AMP-activated protein kinase signaling in metabolic regulation. , 2006, The Journal of clinical investigation.

[158]  W. Kolch.  Coordinating ERK/MAPK signalling through scaffolds and inhibitors , 2005, Nature Reviews Molecular Cell Biology.

[159]  Jin Zhang,et al.  Signal propagation from membrane messengers to nuclear effectors revealed by reporters of phosphoinositide dynamics and Akt activity. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[160]  C. Burd,et al.  Targeting of the FYVE domain to endosomal membranes is regulated by a histidine switch. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[161]  N. Hay,et al.  The Akt-mTOR tango and its relevance to cancer. , 2005, Cancer cell.

[162]  A. Zetterberg,et al.  Single cell analysis of G1 check points-the relationship between the restriction point and phosphorylation of pRb. , 2005, Experimental cell research.

[163]  Roger Y Tsien,et al.  Spatio-temporal Dynamics of Protein Kinase B/Akt Signaling Revealed by a Genetically Encoded Fluorescent Reporter*♦ , 2005, Journal of Biological Chemistry.

[164]  John D. Scott,et al.  AKAP signalling complexes: focal points in space and time , 2004, Nature Reviews Molecular Cell Biology.

[165]  J. Rossant,et al.  Spatial and temporal patterns of ERK signaling during mouse embryogenesis , 2003, Development.

[166]  Y. Umezawa,et al.  Fluorescent Indicators for Akt/Protein Kinase B and Dynamics of Akt Activity Visualized in Living Cells* , 2003, Journal of Biological Chemistry.

[167]  R. Tsien,et al.  Creating new fluorescent probes for cell biology , 2002, Nature Reviews Molecular Cell Biology.

[168]  W. Koch,et al.  Selective inhibition of heterotrimeric Gs signaling. Targeting the receptor-G protein interface using a peptide minigene encoding the Galpha(s) carboxyl terminus. , 2002, The Journal of biological chemistry.

[169]  R Y Tsien,et al.  Genetically encoded fluorescent reporters of protein tyrosine kinase activities in living cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[170]  G. Tsujimoto,et al.  Novel green fluorescent protein-based ratiometric indicators for monitoring pH in defined intracellular microdomains. , 2001, Biochemical and biophysical research communications.

[171]  J. Shabb.  Physiological substrates of cAMP-dependent protein kinase. , 2001, Chemical reviews.

[172]  R. Lefkowitz,et al.  Expanding roles for beta-arrestins as scaffolds and adapters in GPCR signaling and trafficking. , 2001, Current opinion in cell biology.

[173]  A. Miyawaki,et al.  Circularly permuted green fluorescent proteins engineered to sense Ca2+ , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[174]  S J Remington,et al.  Mechanism and Cellular Applications of a Green Fluorescent Protein-based Halide Sensor* , 2000, The Journal of Biological Chemistry.

[175]  Alexander G. Gray,et al.  The pleckstrin homology domains of protein kinase B and GRP1 (general receptor for phosphoinositides-1) are sensitive and selective probes for the cellular detection of phosphatidylinositol 3,4-bisphosphate and/or phosphatidylinositol 3,4,5-trisphosphate in vivo. , 1999, The Biochemical journal.

[176]  R. Tsien,et al.  Circular permutation and receptor insertion within green fluorescent proteins. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[177]  Gero Miesenböck,et al.  Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins , 1998, Nature.

[178]  F. Giancotti,et al.  Integrin signaling: specificity and control of cell survival and cell cycle progression. , 1997, Current opinion in cell biology.

[179]  Roger Y. Tsien,et al.  Crystal Structure of the Aequorea victoria Green Fluorescent Protein , 1996, Science.

[180]  R. Tsien,et al.  Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer , 1996, Current Biology.

[181]  C. D. dos Remedios,et al.  Fluorescence resonance energy transfer spectroscopy is a reliable "ruler" for measuring structural changes in proteins. Dispelling the problem of the unknown orientation factor. , 1995, Journal of structural biology.

[182]  S. Hell,et al.  Properties of a 4Pi confocal fluorescence microscope , 1992 .

[183]  A. Zetterberg,et al.  Kinetic analysis of regulatory events in G1 leading to proliferation or quiescence of Swiss 3T3 cells. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[184]  A. Pardee,et al.  A restriction point for control of normal animal cell proliferation. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[185]  E. Abbe.  Beiträge zur Theorie des Mikroskops und der mikroskopischen Wahrnehmung , 1873 .

[186]  Holger Erfle,et al.  Super-Resolution Microscopy , 2017, Methods in Molecular Biology.

[187]  Benjamien Moeyaert,et al.  PcSOFI as a smart label-based superresolution microscopy technique. , 2014, Methods in molecular biology.

[188]  橋本 寿之.  Time-lapse imaging of cell cycle dynamics during development in living cardiomyocyte(審査報告) , 2014 .

[189]  Marielle Afanassieff,et al.  A short G1 phase is an intrinsic determinant of naïve embryonic stem cell pluripotency. , 2013, Stem cell research.

[190]  K. Sumiyama,et al.  Live imaging of protein kinase activities in transgenic mice expressing FRET biosensors. , 2012, Cell structure and function.

[191]  Jin Zhang,et al.  Visualization of PKA activity in plasma membrane microdomains. , 2011, Molecular bioSystems.

[192]  Massimo Libra,et al.  Akt as a therapeutic target in cancer. , 2008, Expert opinion on therapeutic targets.

[193]  T. Jovin,et al.  FRET imaging , 2003, Nature Biotechnology.

[194]  Roger Y. Tsien,et al.  Creating new fluorescent probes for cell biology , 2003, Nature Reviews Molecular Cell Biology.