PMID- 18350138
OWN - NLM
STAT- MEDLINE
DCOM- 20080808
LR  - 20211020
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 3
IP  - 3
DP  - 2008 Mar 19
TI  - Characterization and subcellular targeting of GCaMP-type genetically-encoded 
      calcium indicators.
PG  - e1796
LID - 10.1371/journal.pone.0001796 [doi]
LID - e1796
AB  - Genetically-encoded calcium indicators (GECIs) hold the promise of monitoring 
      [Ca(2+)] in selected populations of neurons and in specific cellular 
      compartments. Relating GECI fluorescence to neuronal activity requires 
      quantitative characterization. We have characterized a promising new 
      genetically-encoded calcium indicator-GCaMP2-in mammalian pyramidal neurons. 
      Fluorescence changes in response to single action potentials (17+/-10% DeltaF/F 
      [mean+/-SD]) could be detected in some, but not all, neurons. Trains of 
      high-frequency action potentials yielded robust responses (302+/-50% for trains 
      of 40 action potentials at 83 Hz). Responses were similar in acute brain slices 
      from in utero electroporated mice, indicating that long-term expression did not 
      interfere with GCaMP2 function. Membrane-targeted versions of GCaMP2 did not 
      yield larger signals than their non-targeted counterparts. We further targeted 
      GCaMP2 to dendritic spines to monitor Ca(2+) accumulations evoked by activation 
      of synaptic NMDA receptors. We observed robust DeltaF/F responses (range: 
      37%-264%) to single spine uncaging stimuli that were correlated with NMDA 
      receptor currents measured through a somatic patch pipette. One major drawback of 
      GCaMP2 was its low baseline fluorescence. Our results show that GCaMP2 is 
      improved from the previous versions of GCaMP and may be suited to detect bursts 
      of high-frequency action potentials and synaptic currents in vivo.
FAU - Mao, Tianyi
AU  - Mao T
AD  - Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States of 
      America.
FAU - O'Connor, Daniel H
AU  - O'Connor DH
FAU - Scheuss, Volker
AU  - Scheuss V
FAU - Nakai, Junichi
AU  - Nakai J
FAU - Svoboda, Karel
AU  - Svoboda K
LA  - eng
GR  - Howard Hughes Medical Institute/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20080319
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Action Potentials
MH  - Animals
MH  - Calcium/*metabolism
MH  - Hippocampus/metabolism/physiology
MH  - Humans
MH  - In Vitro Techniques
MH  - Rats
MH  - Subcellular Fractions/*metabolism
PMC - PMC2262942
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2008/03/20 09:00
MHDA- 2008/08/09 09:00
PMCR- 2008/03/19
CRDT- 2008/03/20 09:00
PHST- 2007/12/19 00:00 [received]
PHST- 2008/02/13 00:00 [accepted]
PHST- 2008/03/20 09:00 [pubmed]
PHST- 2008/08/09 09:00 [medline]
PHST- 2008/03/20 09:00 [entrez]
PHST- 2008/03/19 00:00 [pmc-release]
AID - 07-PONE-RA-03091R1 [pii]
AID - 10.1371/journal.pone.0001796 [doi]
PST - epublish
SO  - PLoS One. 2008 Mar 19;3(3):e1796. doi: 10.1371/journal.pone.0001796.