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 B. Hogue, Lynn W. Enquist, Mala Murthy, Samuel S.H. Wang

Research output: Contribution to journalArticle

73 Scopus citations

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

All Science Journal Classification (ASJC) codes

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

Fingerprint Dive into the research topics of 'Fast GCaMPs for improved tracking of neuronal activity'. Together they form a unique fingerprint.

  • Cite this

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