Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP

GCaMP, one popular type of genetically-encoded Ca 2+ indicator, has been associated with various side-effects. Here we unveil the intrinsic problem prevailing over different versions and applications, showing that GCaMP containing CaM (calmodulin) interferes with both gating and signaling of L-type...

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Vydáno v:Nature communications Ročník 9; číslo 1; s. 1504 - 18
Hlavní autoři: Yang, Yaxiong, Liu, Nan, He, Yuanyuan, Liu, Yuxia, Ge, Lin, Zou, Linzhi, Song, Sen, Xiong, Wei, Liu, Xiaodong
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 17.04.2018
Nature Publishing Group
Nature Portfolio
Témata:
14/1
14/19
14/34
14/35
14/63
42/109
631/1647/245/2225
631/378/2586
631/45/269/1146
631/80/86/1999
631/80/86/2372
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Animals
Biosensing Techniques - methods
Calcium - chemistry
Calcium - metabolism
Calcium channels
Calcium channels (L-type)
Calcium channels (voltage-gated)
Calcium Channels, L-Type - metabolism
Calcium imaging
Calcium ions
Calcium Signaling
Calcium signalling
Calcium-binding protein
Calmodulin
Calmodulin - chemistry
Calmodulin - genetics
Calmodulin - metabolism
Channel gating
Coupling (molecular)
Fluorescent Dyes - chemistry
Gene expression
Genetic Engineering - methods
Green Fluorescent Proteins - chemistry
Green Fluorescent Proteins - genetics
HEK293 Cells
Humanities and Social Sciences
Humans
Intravital Microscopy - methods
Ion Channel Gating
Mice
Mice, Inbred ICR
Molecular Imaging - methods
Morphogenesis
multidisciplinary
Neurons
Patch-Clamp Techniques
Primary Cell Culture
Protein Binding
Science
Science (multidisciplinary)
Side effects
Transcription
ISSN:2041-1723, 2041-1723
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Popis
Shrnutí:GCaMP, one popular type of genetically-encoded Ca 2+ indicator, has been associated with various side-effects. Here we unveil the intrinsic problem prevailing over different versions and applications, showing that GCaMP containing CaM (calmodulin) interferes with both gating and signaling of L-type calcium channels (Ca V 1). GCaMP acts as an impaired apoCaM and Ca 2+ /CaM, both critical to Ca V 1, which disrupts Ca 2+ dynamics and gene expression. We then design and implement GCaMP-X, by incorporating an extra apoCaM-binding motif, effectively protecting Ca V 1-dependent excitation–transcription coupling from perturbations. GCaMP-X resolves the problems of detrimental nuclear accumulation, acute and chronic Ca 2+ dysregulation, and aberrant transcription signaling and cell morphogenesis, while still demonstrating excellent Ca 2+ -sensing characteristics partly inherited from GCaMP. In summary, CaM/Ca V 1 gating and signaling mechanisms are elucidated for GCaMP side-effects, while allowing the development of GCaMP-X to appropriately monitor cytosolic, submembrane or nuclear Ca 2+ , which is also expected to guide the future design of CaM-based molecular tools. The popular genetically-encoded Ca 2+ indicator, GCaMP, has several side-effects. Here the authors show that GCaMP containing CaM interferes with gating and signaling of L-type calcium channels, which disrupts Ca 2+ dynamics and gene expression, and develop GCaMP-X to overcome these limitations.
Bibliografie:ObjectType-Article-1
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ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-018-03719-6