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Related Experiment Video

Updated: Jun 24, 2026

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators
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Imaging cytoplasmic cAMP in mouse brainstem neurons.

S L Mironov1, E Skorova, G Taschenberger

  • 1DFG-Center of Molecular Physiology of the Brain, Department of Neuro- and Sensory Physiology, Humboldtallee 23, Georg-August-University, 37073 Göttingen, Germany. smirono@gwdg.de

BMC Neuroscience
|March 31, 2009
PubMed
Summary

Researchers developed a novel neuron-specific cyclic adenosine monophosphate (cAMP) probe for real-time imaging in living brain tissue. This tool reveals the interplay between calcium and cAMP signaling in neuronal function and plasticity.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Cell Signaling

Background:

  • Cyclic adenosine monophosphate (cAMP) is a crucial second messenger in neuronal function, often regulated by intracellular calcium (Ca2+).
  • Direct measurement of cAMP dynamics in living vertebrate neurons has been a significant challenge in neuroscience research.

Purpose of the Study:

  • To develop and validate a neuron-specific fluorescent probe for real-time cAMP imaging in living vertebrate neuronal tissue.
  • To investigate the dynamic interplay and crosstalk between Ca2+ and cAMP signaling pathways in neurons.

Main Methods:

  • Development of a neuron-specific cAMP sensor (Epac1-camps) using viral gene transfer for expression in organotypic brainstem slices.
  • Utilized dual-wavelength measurements for absolute quantification of intracellular cAMP ([cAMP]i) levels.

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  • Stimulated neurons via membrane depolarization, G-protein-coupled receptor activation, and pharmacological modulation of adenylate cyclase and phosphodiesterase.

    • Main Results:

    • Successfully expressed the Epac1-camps probe in neuronal cell bodies and neurites, enabling visualization of cAMP distribution.
    • Measured physiologically relevant [cAMP]i changes in response to various stimuli, including membrane depolarization and Ca2+ release.
    • Demonstrated Ca2+-dependent cAMP transients, showing potentiation with phosphodiesterase inhibition and suppression with adenylate cyclase inhibition.

    Conclusions:

    • The developed Epac1-camps probe is a robust tool for studying cAMP dynamics and neuronal plasticity in living brain preparations.
    • The study reveals a synergistic interaction between Ca2+ and cAMP signaling, highlighting their cooperative roles in neuronal function.
    • The probe's characteristics, including strong fluorescence and photobleaching resistance, facilitate advanced neuroscience research.