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Copyright Nature Publishing Group Jul 2013

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.

Details

Title
Fast GCaMPs for improved tracking of neuronal activity
Author
Sun, Xiaonan R; Badura, Aleksandra; Pacheco, Diego A; Lynch, Laura A; Schneider, Eve R; Taylor, Matthew P; Hogue, Ian B; Enquist, Lynn W; Murthy, Mala; Wang, Samuel S -h
Pages
2170
Publication year
2013
Publication date
Jul 2013
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
1400664537
Copyright
Copyright Nature Publishing Group Jul 2013