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@ARTICLE{Kempmann:908173,
author = {Kempmann, Annika and Gensch, Thomas and Offenhäusser,
Andreas and Tihaa, Irina and Maybeck, Vanessa and Balfanz,
Sabine and Baumann, Arnd},
title = {{T}he {F}unctional {C}haracterization of {GC}a{MP}3.0
{V}ariants {S}pecifically {T}argeted to {S}ubcellular
{D}omains},
journal = {International journal of molecular sciences},
volume = {23},
number = {12},
issn = {1422-0067},
address = {Basel},
publisher = {Molecular Diversity Preservation International},
reportid = {FZJ-2022-02431},
pages = {6593 -},
year = {2022},
abstract = {Calcium (Ca2+) ions play a pivotal role in physiology and
cellular signaling. The intracellular Ca2+ concentration
([Ca2+]i) is about three orders of magnitude lower than the
extracellular concentration, resulting in a steep
transmembrane concentration gradient. Thus, the spatial and
the temporal dynamics of [Ca2+]i are ideally suited to
modulate Ca2+-mediated cellular responses to external
signals. A variety of highly sophisticated methods have been
developed to gain insight into cellular Ca2+ dynamics. In
addition to electrophysiological measurements and the
application of synthetic dyes that change their fluorescent
properties upon interaction with Ca2+, the introduction and
the ongoing development of genetically encoded Ca2+
indicators (GECI) opened a new era to study Ca2+-driven
processes in living cells and organisms. Here, we have
focused on one well-established GECI, i.e., GCaMP3.0. We
have systematically modified the protein with sequence
motifs, allowing localization of the sensor in the nucleus,
in the mitochondrial matrix, at the mitochondrial outer
membrane, and at the plasma membrane. The individual
variants and a cytosolic version of GCaMP3.0 were
overexpressed and purified from E. coli cells to study their
biophysical properties in solution. All versions were
examined to monitor Ca2+ signaling in stably transfected
cell lines and in primary cortical neurons transduced with
recombinant Adeno-associated viruses (rAAV). In this
comparative study, we provide evidence for a robust approach
to reliably trace Ca2+ signals at the (sub)-cellular level
with pronounced temporal resolution},
cin = {IBI-1 / IBI-3},
ddc = {540},
cid = {I:(DE-Juel1)IBI-1-20200312 / I:(DE-Juel1)IBI-3-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {35743038},
UT = {WOS:000816484300001},
doi = {10.3390/ijms23126593},
url = {https://juser.fz-juelich.de/record/908173},
}