Fast GCaMPs for improved tracking of neuronal activity

Xiaonan R. Sun, Aleksandra Badura, Diego A. Pacheco, Laura A. Lynch, Eve R. Schneider, Matthew P. Taylor, Ian Hogue, Lynn W. Enquist, Mala Murthy, Samuel S.H. Wang

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

The use of genetically encodable calcium indicator proteins to monitor neuronal activity is hampered by slow response times and a narrow Ca2+-sensitive range. Here we identify three performance-limiting features of GCaMP3, a popular genetically encodable calcium indicator protein. First, we find that affinity is regulated by the calmodulin domain's Ca2+-chelating residues. Second, we find that off-responses to Ca2+ are rate-limited by dissociation of the RS20 domain from calmodulin's hydrophobic pocket. Third, we find that on-responses are limited by fast binding to the N-lobe at high Ca2+ and by slow binding to the C-lobe at lower Ca2+. We develop Fast-GCaMPs, which have up to 20-fold accelerated off-responses and show that they have a 200-fold range of KD, allowing coexpression of multiple variants to span an expanded range of Ca2+ concentrations. Finally, we show that Fast-GCaMPs track natural song in Drosophila auditory neurons and generate rapid responses in mammalian neurons, supporting the utility of our approach.

Original languageEnglish (US)
Article number2170
JournalNature Communications
Volume4
DOIs
StatePublished - Jul 18 2013
Externally publishedYes

Fingerprint

Calmodulin
Neurons
Calcium
calmodulin
Music
Chelation
Reaction Time
Drosophila
Proteins
neurons
lobes
calcium
proteins
affinity
dissociation

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Sun, X. R., Badura, A., Pacheco, D. A., Lynch, L. A., Schneider, E. R., Taylor, M. P., ... Wang, S. S. H. (2013). Fast GCaMPs for improved tracking of neuronal activity. Nature Communications, 4, [2170]. https://doi.org/10.1038/ncomms3170

Fast GCaMPs for improved tracking of neuronal activity. / Sun, Xiaonan R.; Badura, Aleksandra; Pacheco, Diego A.; Lynch, Laura A.; Schneider, Eve R.; Taylor, Matthew P.; Hogue, Ian; Enquist, Lynn W.; Murthy, Mala; Wang, Samuel S.H.

In: Nature Communications, Vol. 4, 2170, 18.07.2013.

Research output: Contribution to journalArticle

Sun, XR, Badura, A, Pacheco, DA, Lynch, LA, Schneider, ER, Taylor, MP, Hogue, I, Enquist, LW, Murthy, M & Wang, SSH 2013, 'Fast GCaMPs for improved tracking of neuronal activity', Nature Communications, vol. 4, 2170. https://doi.org/10.1038/ncomms3170
Sun XR, Badura A, Pacheco DA, Lynch LA, Schneider ER, Taylor MP et al. Fast GCaMPs for improved tracking of neuronal activity. Nature Communications. 2013 Jul 18;4. 2170. https://doi.org/10.1038/ncomms3170
Sun, Xiaonan R. ; Badura, Aleksandra ; Pacheco, Diego A. ; Lynch, Laura A. ; Schneider, Eve R. ; Taylor, Matthew P. ; Hogue, Ian ; Enquist, Lynn W. ; Murthy, Mala ; Wang, Samuel S.H. / Fast GCaMPs for improved tracking of neuronal activity. In: Nature Communications. 2013 ; Vol. 4.
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