Blue fluorescent cGMP sensor for multiparameter fluorescence imaging

Yusuke Niino, Kohji Hotta, Kotaro Oka

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Cyclic GMP (cGMP) regulates many physiological processes by cooperating with the other signaling molecules such as cyclic AMP (cAMP) and Ca 2+. Genetically encoded sensors for cGMP have been developed based on fluorescence resonance energy transfer (FRET) between fluorescent proteins. However, to analyze the dynamic relationship among these second messengers, combined use of existing sensors in a single cell is inadequate because of the significant spectral overlaps. A single wavelength indicator is an effective alternative to avoid this problem, but color variants of a single fluorescent protein-based biosensor are limited. In this study, to construct a new color fluorescent sensor, we converted the FRET-based sensor into a single wavelength indicator using a dark FRET acceptor. We developed a blue fluorescent cGMP biosensor, which is spectrally compatible with a FRET-based cAMP sensor using cyan and yellow fluorescent proteins (CFP/YFP). We cotransfected them and loaded a red fluorescent probe for Ca2+ into cells, and accomplished triple-parameter fluorescence imaging of these cyclic nucleotides and Ca 2+, confirming the applicability of this combination to individually monitor their dynamics in a single cell. This blue fluorescent sensor and the approach using this FRET pair would be useful for multiparameter fluorescence imaging to understand complex signal transduction networks.

Original languageEnglish
Article numbere9164
JournalPLoS One
Volume5
Issue number2
DOIs
Publication statusPublished - 2010 Feb 11

Fingerprint

cyclic GMP
Fluorescence Resonance Energy Transfer
Optical Imaging
Cyclic GMP
sensors (equipment)
Fluorescence
energy transfer
image analysis
fluorescence
Imaging techniques
Sensors
Biosensing Techniques
Cyclic AMP
biosensors
Biosensors
cyclic AMP
Color
Physiological Phenomena
wavelengths
Cyclic Nucleotides

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Blue fluorescent cGMP sensor for multiparameter fluorescence imaging. / Niino, Yusuke; Hotta, Kohji; Oka, Kotaro.

In: PLoS One, Vol. 5, No. 2, e9164, 11.02.2010.

Research output: Contribution to journalArticle

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