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Presentation Abstract
Program#/Poster#: 590.12/XX18
Presentation Title: In vivo profiling of astrocytic calcium excitability and downstream vascular effects:
Fluorescence intensity and lifetime measurements
Location: Hall F-J
Presentation time: Tuesday, Oct 16, 2012, 11:00 AM -12:00 PM
Authors: *H. UHLIROVA1, P. A. SAISAN2, E. S. NORHEIM2,4, Q. CHENG2, K. L.
WELDY2, S. L. GRATIY1, G. T. EINEVOLL4, A. M. DALE3, A. DEVOR3,5; 1Radiology, 2Neurosciences, 3Neurosciences & Radiology, Univ. of California San
Diego, La Jolla, CA; 4Dept. of Mathematical Sci. and Technol. and Ctr. for
Integrative Genet. (CIGENE), Norwegian Univ. of Life Sci., Oslo, Norway; 5MGH,
Harvard Med. Sch., Charlestown, MA
Abstract: Astrocytes express a number of metabotropic receptors for messenger molecules,including glutamate and ATP, and can exhibit elevations of intracellular calcium upon
their activation. An increase in astrocytic calcium, in turn, can lead to release ofgliotransmitters implicated in neuroglial and gliovascular interactions. In this study, we
quantitatively assessed in vivo astrocytic calcium dynamics and their downstream
vascular effects in response to local micropharmacologocal stimulations. Time-
resolved astrocytic calcium concentration (in nM) was measured through
fluorescence lifetime imaging microscopy (FLIM) of a popular calcium indicator,
Oregon Green BAPTA-1 AM (1-3). To achieve a comparable temporal resolution
to the widely used fluorescence intensity calcium imaging (~10 Hz), we implemented
user-defined scanning trajectories aiming to collect sufficient numbers of photons
from a region of interest (e.g., a single astrocytic cell body) per time-point (Figure
1A). Our data show large increases in astrocytic intracellular calcium (>150 nM at
the peak) in the absence of arteriolar dilation, and arteriolar dilation in the absence ofthe astrocytic calcium increase. In addition, an increase in astrocytic intracellular
calcium concentration measured by FLIM exceeded the ΔF/F obtained by
fluorescence intensity imaging (Figure 1B). This effect might result from the presence
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of indicator molecules in intracellular calcium stores isolated from the cytosolic
calcium changes. Figure 1. Astrocytic dynamics of the absolute calcium concentration
in vivo. A. Field of view with 3 astrocytes (yellow). Scan trajectory for FLIM
measurement is shown on the right. B-C. FLIM (B) and intensity (C) single-trial
time-courses in response to a micropuff of 1 mM ATP. Red vertical lines indicate the
timing of the puff.1. Wilms CD, et al. (2006) Cell Calcium 40(1):73-79.
2. Lattarulo C, et al.(2011) Methods Mol Biol 793:377-389.
3. Gersbach M, et al. (2009) Opt Lett 34(3):362-364.
Disclosures: H. Uhlirova: None. P.A. Saisan: None. E.S. Norheim: None. Q. Cheng:None. K.L. Weldy: None. S.L. Gratiy: None. G.T. Einevoll: None. A.M. Dale:
None. A. Devor: None.
Keyword(s): 2-photon microscopy
ASTROCYTE
BLOOD FLOW
Support: NIH Grant NS051188
NIH Grant NS057198
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NIH Grant EB009118
NIH Grant EB00790
European Regional Development Fund CEITEC
[Authors]. [Abstract Title]. Program No. XXX.XX. 2012 Neuroscience MeetingPlanner. New Orleans, LA: Society for Neuroscience, 2012. Online.
2012 Copyright by the Society for Neuroscience all rights reserved. Permission to
republish any abstract or part of any abstract in any form must be obtained in writingby SfN office prior to publication.