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Related Concept Videos

Intracellular Signaling Cascades01:24

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Once a ligand binds to a receptor, the signal is transmitted through the membrane and into the cytoplasm. The continuation of a signal in this manner is called signal transduction. Signal transduction only occurs with cell-surface receptors, which cannot interact with most components of the cell, such as DNA. Only internal receptors can interact directly with DNA in the nucleus to initiate protein synthesis. When a ligand binds to its receptor, conformational changes occur that affect the...
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Measurement of 3-Dimensional cAMP Distributions in Living Cells using 4-Dimensional x, y, z, and &lambda; Hyperspectral FRET Imaging and Analysis
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Interrogating cyclic AMP signaling using optical approaches.

Jason Y Jiang1, Jeffrey L Falcone1, Silvana Curci1

  • 1VA Boston Healthcare System and the Dept. of Surgery, Brigham & Women's Hospital and Harvard Medical School, 1400 VFW PKW, West Roxbury, MA 02132, USA.

Cell Calcium
|March 10, 2017
PubMed
Summary
This summary is machine-generated.

Optical reporters offer precise measurement of cyclic adenosine monophosphate (cAMP) signaling in single cells. Genetically encoded fluorescent sensors have revolutionized cAMP research, enabling detailed investigation of cellular dynamics.

Keywords:
EpacFRETFluorescent proteinsPKAcAMP signaling microdomains

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Area of Science:

  • Cellular signaling
  • Molecular biology
  • Biophysics

Background:

  • Optical reporters for cyclic adenosine monophosphate (cAMP) are crucial for understanding cellular signaling dynamics.
  • Early methods required large cell populations; optical sensors allow single-cell and subcellular measurements.
  • Genetically encoded fluorescent sensors have significantly advanced cAMP research since their introduction.

Purpose of the Study:

  • To review the development and advancements in optical reporters for cAMP signaling.
  • To highlight the advantages of optical sensors over traditional cAMP measurement methods.
  • To discuss current and potential future strategies for optical cAMP sensing.

Main Methods:

  • Development of Förster Resonance Energy Transfer (FRET)-based sensors, initially using PKA subunits.
  • Engineering of genetically encoded sensors for cellular introduction via transfection.
  • Utilization of Epac protein as a common cAMP binding platform in FRET sensors.
  • Exploration of alternative optical sensing strategies including BRET and translocation reporters.

Main Results:

  • Genetically encoded FRET sensors, particularly those based on Epac, are widely used.
  • Various cAMP binding proteins and engineered fluorescent proteins have been employed in sensor design.
  • Optical reporters provide high spatial and temporal resolution for cAMP dynamics.
  • New sensor designs continue to emerge, improving sensitivity and specificity.

Conclusions:

  • Optical reporters have fundamentally advanced the study of cAMP signaling dynamics.
  • Genetically encoded sensors offer significant advantages for investigating cAMP in specific cellular locations.
  • Ongoing innovation in fluorescent proteins and binding domains promises further refinement of cAMP optical reporters.