PMID- 25490346
OWN - NLM
STAT- MEDLINE
DCOM- 20150630
LR  - 20220309
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 93
DP  - 2014 Nov 19
TI  - Imaging intracellular Ca(2)(+) signals in striatal astrocytes from adult mice using 
      genetically-encoded calcium indicators.
PG  - e51972
LID - 10.3791/51972 [doi]
LID - 51972
AB  - Astrocytes display spontaneous intracellular Ca(2+) concentration fluctuations 
      ([Ca(2+)]i) and in several settings respond to neuronal excitation with enhanced 
      [Ca(2+)]i signals. It has been proposed that astrocytes in turn regulate neurons 
      and blood vessels through calcium-dependent mechanisms, such as the release of 
      signaling molecules. However, [Ca(2+)]i imaging in entire astrocytes has only 
      recently become feasible with genetically encoded calcium indicators (GECIs) such 
      as the GCaMP series. The use of GECIs in astrocytes now provides opportunities to 
      study astrocyte [Ca(2+)]i signals in detail within model microcircuits such as 
      the striatum, which is the largest nucleus of the basal ganglia. In the present 
      report, detailed surgical methods to express GECIs in astrocytes in vivo, and 
      confocal imaging approaches to record [Ca(2+)]i signals in striatal astrocytes in 
      situ, are described. We highlight precautions, necessary controls and tests to 
      determine if GECI expression is selective for astrocytes and to evaluate signs of 
      overt astrocyte reactivity. We also describe brain slice and imaging conditions 
      in detail that permit reliable [Ca(2+)]i imaging in striatal astrocytes in situ. 
      The use of these approaches revealed the entire territories of single striatal 
      astrocytes and spontaneous [Ca(2+)]i signals within their somata, branches and 
      branchlets. The further use and expansion of these approaches in the striatum 
      will allow for the detailed study of astrocyte [Ca(2+)]i signals in the striatal 
      microcircuitry.
FAU - Jiang, Ruotian
AU  - Jiang R
AD  - Department of Physiology, University of California Los Angeles.
FAU - Haustein, Martin D
AU  - Haustein MD
AD  - Department of Physiology, University of California Los Angeles.
FAU - Sofroniew, Michael V
AU  - Sofroniew MV
AD  - Department of Neurobiology, University of California Los Angeles.
FAU - Khakh, Baljit S
AU  - Khakh BS
AD  - Department of Physiology, University of California Los Angeles; Department of 
      Neurobiology, University of California Los Angeles; BKhakh@mednet.ucla.edu.
LA  - eng
GR  - R01 MH099559/MH/NIMH NIH HHS/United States
GR  - MH104069/MH/NIMH NIH HHS/United States
GR  - NS060677/NS/NINDS NIH HHS/United States
GR  - R01 NS060677/NS/NINDS NIH HHS/United States
GR  - MH099559/MH/NIMH NIH HHS/United States
GR  - DP1 MH104069/MH/NIMH NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20141119
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Receptors, Calcium-Sensing)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Adenoviridae/genetics
MH  - Animals
MH  - Astrocytes/*chemistry/metabolism
MH  - Calcium/*analysis/metabolism
MH  - Calcium Signaling/*physiology
MH  - Corpus Striatum/*chemistry/cytology/metabolism
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Receptors, Calcium-Sensing/genetics/metabolism
PMC - PMC4354287
EDAT- 2014/12/10 06:00
MHDA- 2015/07/01 06:00
PMCR- 2016/11/19
CRDT- 2014/12/10 06:00
PHST- 2014/12/10 06:00 [entrez]
PHST- 2014/12/10 06:00 [pubmed]
PHST- 2015/07/01 06:00 [medline]
PHST- 2016/11/19 00:00 [pmc-release]
AID - 51972 [pii]
AID - 10.3791/51972 [doi]
PST - epublish
SO  - J Vis Exp. 2014 Nov 19;(93):e51972. doi: 10.3791/51972.