Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging.
Animals
Brain
/ metabolism
Caenorhabditis elegans
/ metabolism
Calcium
/ metabolism
Fluorescent Dyes
Green Fluorescent Proteins
/ chemistry
HeLa Cells
Humans
Indicators and Reagents
Luminescent Proteins
/ chemistry
Mice
Myocytes, Cardiac
/ metabolism
Neurons
/ metabolism
Optical Imaging
/ methods
Optogenetics
Protein Engineering
Spectroscopy, Near-Infrared
Xenopus laevis
/ metabolism
Journal
PLoS biology
ISSN: 1545-7885
Titre abrégé: PLoS Biol
Pays: United States
ID NLM: 101183755
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
14
04
2020
accepted:
29
10
2020
revised:
08
12
2020
pubmed:
25
11
2020
medline:
5
1
2021
entrez:
24
11
2020
Statut:
epublish
Résumé
Near-infrared (NIR) genetically encoded calcium ion (Ca2+) indicators (GECIs) can provide advantages over visible wavelength fluorescent GECIs in terms of reduced phototoxicity, minimal spectral cross talk with visible light excitable optogenetic tools and fluorescent probes, and decreased scattering and absorption in mammalian tissues. Our previously reported NIR GECI, NIR-GECO1, has these advantages but also has several disadvantages including lower brightness and limited fluorescence response compared to state-of-the-art visible wavelength GECIs, when used for imaging of neuronal activity. Here, we report 2 improved NIR GECI variants, designated NIR-GECO2 and NIR-GECO2G, derived from NIR-GECO1. We characterized the performance of the new NIR GECIs in cultured cells, acute mouse brain slices, and Caenorhabditis elegans and Xenopus laevis in vivo. Our results demonstrate that NIR-GECO2 and NIR-GECO2G provide substantial improvements over NIR-GECO1 for imaging of neuronal Ca2+ dynamics.
Identifiants
pubmed: 33232322
doi: 10.1371/journal.pbio.3000965
pii: PBIOLOGY-D-20-00994
pmc: PMC7723245
doi:
Substances chimiques
Fluorescent Dyes
0
Indicators and Reagents
0
Luminescent Proteins
0
Green Fluorescent Proteins
147336-22-9
Calcium
SY7Q814VUP
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
e3000965Subventions
Organisme : CIHR
ID : MOP-123514
Pays : Canada
Organisme : CIHR
ID : FS-154310
Pays : Canada
Organisme : NIDA NIH HHS
ID : R01 DA045549
Pays : United States
Organisme : NIDA NIH HHS
ID : R01 DA029639
Pays : United States
Organisme : Howard Hughes Medical Institute
Pays : United States
Organisme : CIHR
ID : FDN-143238
Pays : Canada
Organisme : NINDS NIH HHS
ID : U01 NS094246
Pays : United States
Organisme : NINDS NIH HHS
ID : RF1 NS113287
Pays : United States
Organisme : NINDS NIH HHS
ID : U01 NS090565
Pays : United States
Organisme : NIMH NIH HHS
ID : R01 MH122971
Pays : United States
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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