PMID- 32344594
OWN - NLM
STAT- MEDLINE
DCOM- 20210201
LR  - 20210201
IS  - 1422-0067 (Electronic)
IS  - 1422-0067 (Linking)
VI  - 21
IP  - 8
DP  - 2020 Apr 24
TI  - FGCaMP7, an Improved Version of Fungi-Based Ratiometric Calcium Indicator for In 
      Vivo Visualization of Neuronal Activity.
LID - 10.3390/ijms21083012 [doi]
LID - 3012
AB  - Genetically encoded calcium indicators (GECIs) have become a widespread tool for 
      the visualization of neuronal activity. As compared to popular GCaMP GECIs, the 
      FGCaMP indicator benefits from calmodulin and M13-peptide from the fungi 
      Aspergillus niger and Aspergillus fumigatus, which prevent its interaction with 
      the intracellular environment. However, FGCaMP exhibits a two-phase fluorescence 
      behavior with the variation of calcium ion concentration, has moderate 
      sensitivity in neurons (as compared to the GCaMP6s indicator), and has not been 
      fully characterized in vitro and in vivo. To address these limitations, we 
      developed an enhanced version of FGCaMP, called FGCaMP7. FGCaMP7 preserves the 
      ratiometric phenotype of FGCaMP, with a 3.1-fold larger ratiometric dynamic range 
      in vitro. FGCaMP7 demonstrates 2.7- and 8.7-fold greater photostability compared 
      to mEGFP and mTagBFP2 fluorescent proteins in vitro, respectively. The 
      ratiometric response of FGCaMP7 is 1.6- and 1.4-fold higher, compared to the 
      intensiometric response of GCaMP6s, in non-stimulated and stimulated neuronal 
      cultures, respectively. We reveal the inertness of FGCaMP7 to the intracellular 
      environment of HeLa cells using its truncated version with a deleted M13-like 
      peptide; in contrast to the similarly truncated variant of GCaMP6s. We 
      characterize the crystal structure of the parental FGCaMP indicator. Finally, we 
      test the in vivo performance of FGCaMP7 in mouse brain using a two-photon 
      microscope and an NVista miniscope; and in zebrafish using two-color ratiometric 
      confocal imaging.
FAU - Barykina, Natalia V
AU  - Barykina NV
AUID- ORCID: 0000-0001-5835-8388
AD  - Laboratory for Neurobiology of Memory, P.K. Anokhin Research Institute of Normal 
      Physiology, 125315 Moscow, Russia.
FAU - Sotskov, Vladimir P
AU  - Sotskov VP
AUID- ORCID: 0000-0002-1729-5744
AD  - Institute for Advanced Brain Studies, M.V. Lomonosov Moscow State University, 
      119991 Moscow, Russia.
FAU - Gruzdeva, Anna M
AU  - Gruzdeva AM
AUID- ORCID: 0000-0001-5691-9057
AD  - Institute for Advanced Brain Studies, M.V. Lomonosov Moscow State University, 
      119991 Moscow, Russia.
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
AD  - Sensorimotor Control Research Group, Max Planck Institute of Neurobiology, 82152 
      Martinsried, Germany.
FAU - Wu, You Kure
AU  - Wu YK
AD  - Sensorimotor Control Research Group, Max Planck Institute of Neurobiology, 82152 
      Martinsried, Germany.
FAU - Portugues, Ruben
AU  - Portugues R
AD  - Sensorimotor Control Research Group, Max Planck Institute of Neurobiology, 82152 
      Martinsried, Germany.
AD  - Institute of Neuroscience, Technical University of Munich, 80802 Munich, Germany.
AD  - Munich Cluster for Systems Neurology (SyNergy), Munich, Germany.
FAU - Subach, Oksana M
AU  - Subach OM
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
FAU - Chefanova, Elizaveta S
AU  - Chefanova ES
AD  - Department of NBIC-technologies, Moscow Institute of Physics and Technology, 
      123182 Moscow, Russia.
FAU - Plusnin, Viktor V
AU  - Plusnin VV
AUID- ORCID: 0000-0001-9784-8283
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
AD  - Department of NBIC-technologies, Moscow Institute of Physics and Technology, 
      123182 Moscow, Russia.
FAU - Ivashkina, Olga I
AU  - Ivashkina OI
AD  - Laboratory for Neurobiology of Memory, P.K. Anokhin Research Institute of Normal 
      Physiology, 125315 Moscow, Russia.
AD  - Institute for Advanced Brain Studies, M.V. Lomonosov Moscow State University, 
      119991 Moscow, Russia.
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
FAU - Anokhin, Konstantin V
AU  - Anokhin KV
AD  - Laboratory for Neurobiology of Memory, P.K. Anokhin Research Institute of Normal 
      Physiology, 125315 Moscow, Russia.
AD  - Institute for Advanced Brain Studies, M.V. Lomonosov Moscow State University, 
      119991 Moscow, Russia.
FAU - Vlaskina, Anna V
AU  - Vlaskina AV
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
FAU - Korzhenevskiy, Dmitry A
AU  - Korzhenevskiy DA
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
FAU - Nikolaeva, Alena Y
AU  - Nikolaeva AY
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
FAU - Boyko, Konstantin M
AU  - Boyko KM
AUID- ORCID: 0000-0001-8229-189X
AD  - Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian 
      Academy of Sciences, 119071 Moscow, Russia.
FAU - Rakitina, Tatiana V
AU  - Rakitina TV
AUID- ORCID: 0000-0002-3096-8659
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
AD  - Laboratory of Hormonal Regulation Proteins, M.M. Shemyakin and Yu.A. Ovchinnikov 
      Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997 
      Moscow, Russia.
FAU - Varizhuk, Anna M
AU  - Varizhuk AM
AD  - Department of Biophysics, Research and Clinical Center of Physical-Chemical 
      Medicine of Federal Medical Biological Agency, 119435 Moscow, Russia.
AD  - Department of Biophysics, Center for Precision Genome Editing and Genetic 
      Technologies for Biomedicine, 119435 Moscow, Russia.
FAU - Pozmogova, Galina E
AU  - Pozmogova GE
AD  - Department of Biophysics, Research and Clinical Center of Physical-Chemical 
      Medicine of Federal Medical Biological Agency, 119435 Moscow, Russia.
AD  - Department of Biophysics, Center for Precision Genome Editing and Genetic 
      Technologies for Biomedicine, 119435 Moscow, Russia.
FAU - Subach, Fedor V
AU  - Subach FV
AUID- ORCID: 0000-0003-2720-7821
AD  - Complex of NBICS Technologies, National Research Center "Kurchatov Institute", 
      123182 Moscow, Russia.
LA  - eng
GR  - numero sign1360 of 25.06.2019/internal grant of the National Research Center Kurchatov 
      Institute/
GR  - 16-15-10323/Russian Science Foundation/
GR  - 19-04-00395/Russian Foundation for Basic Research/
GR  - 17-00-00215/Russian Foundation for Basic Research/
GR  - 20-015-00427/Russian Foundation for Basic Research/
GR  - 19-315-80020/Russian Foundation for Basic Research/
GR  - 18-34-00640/Russian Foundation for Basic Research/
GR  - 17-29-07083/Russian Foundation for Basic Research/
GR  - to K.M.B./Ministry of Science and Higher Education of the Russian Federation/
GR  - 19-34-70004/Russian Foundation for Basic Research/
GR  - 075-15-2019-1669/the Ministry of Science and Higher Education of the Russian 
      Federation/
GR  - EXC 2145 SyNergy - ID 390857198/Deutsche Forschungsgemeinschaft/
PT  - Journal Article
DEP - 20200424
PL  - Switzerland
TA  - Int J Mol Sci
JT  - International journal of molecular sciences
JID - 101092791
RN  - 0 (Calcium-Binding Proteins)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Action Potentials
MH  - Animals
MH  - Calcium/*metabolism
MH  - Calcium-Binding Proteins/genetics/metabolism
MH  - Fungi/genetics
MH  - *Gene Expression
MH  - Genes, Reporter
MH  - HeLa Cells
MH  - Humans
MH  - Mice
MH  - Microscopy, Fluorescence
MH  - Models, Molecular
MH  - *Molecular Imaging/methods
MH  - Neurons/cytology/*metabolism
MH  - Protein Conformation
MH  - Protein Engineering
MH  - Structure-Activity Relationship
MH  - Visual Cortex/physiology
PMC - PMC7215472
OTO - NOTNLM
OT  - FGCaMP
OT  - FGCaMP7
OT  - calcium imaging
OT  - crystal structure
OT  - genetically encoded calcium indicator
OT  - protein engineering
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses, or interpretation of data; in 
      the writing of the manuscript, or in the decision to publish the results.
EDAT- 2020/04/30 06:00
MHDA- 2021/02/02 06:00
PMCR- 2020/04/01
CRDT- 2020/04/30 06:00
PHST- 2020/04/12 00:00 [received]
PHST- 2020/04/22 00:00 [revised]
PHST- 2020/04/23 00:00 [accepted]
PHST- 2020/04/30 06:00 [entrez]
PHST- 2020/04/30 06:00 [pubmed]
PHST- 2021/02/02 06:00 [medline]
PHST- 2020/04/01 00:00 [pmc-release]
AID - ijms21083012 [pii]
AID - ijms-21-03012 [pii]
AID - 10.3390/ijms21083012 [doi]
PST - epublish
SO  - Int J Mol Sci. 2020 Apr 24;21(8):3012. doi: 10.3390/ijms21083012.