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cAMPr: A single-wavelength fluorescent sensor for cyclic AMP.

Christopher R Hackley1, Esteban O Mazzoni1,2, Justin Blau3,2,4

  • 1Department of Biology, New York University (NYU), 100 Washington Square East, New York, NY 10003, USA.

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Summary

Researchers developed cAMPr, a novel genetically encoded fluorescent sensor for cyclic adenosine monophosphate (cAMP). This advanced sensor offers improved quantitative detection of cAMP and enables simultaneous monitoring of cAMP and Ca2+ in real-time cellular studies.

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Area of Science:

  • Molecular Biology
  • Neuroscience
  • Biochemistry

Background:

  • Genetically encoded fluorescent sensors are crucial for real-time, cell-specific measurements of ions and small molecules.
  • Cyclic adenosine monophosphate (cAMP) is a vital signaling molecule across diverse cell types and organisms.

Purpose of the Study:

  • To develop and validate a new single-wavelength genetically encoded sensor for cyclic adenosine monophosphate (cAMP), named cAMPr.
  • To assess the performance of cAMPr in quantitative cAMP detection and its compatibility with advanced imaging techniques.

Main Methods:

  • Development of the cAMPr sensor in bacteria and embryonic stem cells.
  • Validation in mammalian neurons in vitro and in Drosophila circadian pacemaker neurons in vivo.
  • Comparative analysis with existing single-wavelength cAMP sensors.
  • Testing compatibility with single-photon and two-photon imaging modalities.

Main Results:

  • cAMPr demonstrated improved quantitative detection of cAMP abundance compared to other sensors.
  • The sensor is compatible with both single-photon and two-photon imaging.
  • Simultaneous monitoring of Ca2+ and cAMP was achieved in Drosophila brains using cAMPr and RCaMP1h.

Conclusions:

  • cAMPr is a versatile and effective genetically encoded sensor for cAMP.
  • This sensor facilitates advanced real-time studies of cellular signaling pathways, including the simultaneous measurement of cAMP and Ca2+.
  • cAMPr advances the toolkit for molecular and cellular neuroscience research.