title: Optical recording of neuronal activity with a genetically-encoded calcium indicator in anesthetized and freely moving mice.
abstract: Fluorescent calcium (Ca(2+)) indicator proteins (FCIPs) are promising tools for functional imaging of cellular activity in living animals. However, they have still not reached their full potential for in vivo imaging of neuronal activity due to limitations in expression levels, dynamic range, and sensitivity for reporting action potentials. Here, we report that viral expression of the ratiometric Ca(2+) sensor yellow cameleon 3.60 (YC3.60) in pyramidal neurons of mouse barrel cortex enables in vivo measurement of neuronal activity with high dynamic range and sensitivity across multiple spatial scales. By combining juxtacellular recordings and two-photon imaging in vitro and in vivo, we demonstrate that YC3.60 can resolve single action potential (AP)-evoked Ca(2+) transients and reliably reports bursts of APs with negligible saturation. Spontaneous and whisker-evoked Ca(2+) transients were detected in individual apical dendrites and somata as well as in local neuronal populations. Moreover, bulk measurements using wide-field imaging or fiber-optics revealed sensory-evoked YC3.60 signals in large areas of the barrel field. Fiber-optic recordings in particular enabled measurements in awake, freely moving mice and revealed complex Ca(2+) dynamics, possibly reflecting different behavior-related brain states. Viral expression of YC3.60 - in combination with various optical techniques - thus opens a multitude of opportunities for functional studies of the neural basis of animal behavior, from dendrites to the levels of local and large-scale neuronal populations.
authors.affiliation: Department of Neurophysiology, Brain Research Institute, University of Zurich Zurich, Switzerland.
authors.forename: Henry
authors.initials: H
authors.lastname: Lütcke
authors.forename: Masanori
authors.initials: M
authors.lastname: Murayama
authors.forename: Thomas
authors.initials: T
authors.lastname: Hahn
authors.forename: David J
authors.initials: DJ
authors.lastname: Margolis
authors.forename: Simone
authors.initials: S
authors.lastname: Astori
authors.forename: Stephan Meyer
authors.initials: SM
authors.lastname: Zum Alten Borgloh
authors.forename: Werner
authors.initials: W
authors.lastname: Göbel
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.forename: Wannan
authors.initials: W
authors.lastname: Tang
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kügler
authors.forename: Rolf
authors.initials: R
authors.lastname: Sprengel
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
authors.forename: Atsushi
authors.initials: A
authors.lastname: Miyawaki
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Larkum
authors.forename: Fritjof
authors.initials: F
authors.lastname: Helmchen
authors.forename: Mazahir T
authors.initials: MT
authors.lastname: Hasan
references.citation: Proc Natl Acad Sci U S A. 1994 May 24;91(11):5207-11
references.pmid: 8197208
references.citation: Nat Protoc. 2006;1(3):1057-65
references.pmid: 17406387
references.citation: PLoS One. 2007 Jun 20;2(6):e533
references.pmid: 17579707
references.citation: Chem Biol. 2006 May;13(5):521-30
references.pmid: 16720273
references.citation: Annu Rev Pharmacol Toxicol. 2003;43:413-39
references.pmid: 12359866
references.citation: Nature. 1997 Jan 9;385(6612):161-5
references.pmid: 8990119
references.citation: J Biosci Bioeng. 2008 Feb;105(2):106-9
references.pmid: 18343335
references.citation: J Neurophysiol. 2007 Dec;98(6):3770-9
references.pmid: 17898141
references.citation: Chem Rev. 2008 May;108(5):1550-64
references.pmid: 18447377
references.citation: Physiology (Bethesda). 2007 Dec;22:358-65
references.pmid: 18073408
references.citation: Neuron. 2008 Jul 31;59(2):214-25
references.pmid: 18667150
references.citation: Nat Methods. 2004 Oct;1(1):31-7
references.pmid: 15782150
references.citation: J Physiol. 1995 Sep 15;487 ( Pt 3):583-600
references.pmid: 8544123
references.citation: Nature. 2009 Feb 26;457(7233):1137-41
references.pmid: 19151696
references.citation: J Neurosci. 2000 May 1;20(9):3354-68
references.pmid: 10777798
references.citation: Nat Neurosci. 2010 Jan;13(1):133-40
references.pmid: 20023653
references.citation: Nat Methods. 2008 Sep;5(9):797-804
references.pmid: 19160514
references.citation: Nat Rev Neurosci. 2008 Mar;9(3):195-205
references.pmid: 18270513
references.citation: Neuron. 2006 Mar 16;49(6):877-87
references.pmid: 16543135
references.citation: Nat Neurosci. 2005 Aug;8(8):988-90
references.pmid: 16007081
references.citation: Curr Opin Neurobiol. 2009 Oct;19(5):520-9
references.pmid: 19854041
references.citation: J Neurosci. 2007 Nov 28;27(48):13316-28
references.pmid: 18045926
references.citation: Nat Methods. 2005 Dec;2(12):932-40
references.pmid: 16299478
references.citation: Annu Rev Neurosci. 2009;32:435-506
references.pmid: 19555292
references.citation: J Neurosci. 2003 Sep 17;23(24):8558-67
references.pmid: 13679425
references.citation: Gene Ther. 2007 Nov;14(21):1534-6
references.pmid: 17713567
references.citation: PLoS One. 2008 Apr 16;3(4):e2005
references.pmid: 18414675
references.citation: Am J Hum Genet. 2007 Feb;80(2):291-7
references.pmid: 17236133
references.citation: Nat Methods. 2009 Dec;6(12):875-81
references.pmid: 19898485
references.citation: Adv Biochem Eng Biotechnol. 2005;95:1-15
references.pmid: 16080263
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):14063-8
references.pmid: 16157876
references.citation: J Neurosci. 2008 Jul 9;28(28):7025-30
references.pmid: 18614669
references.citation: Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10554-9
references.pmid: 15247428
references.citation: PLoS Biol. 2004 Jun;2(6):e163
references.pmid: 15208716
references.citation: J Neurosci Methods. 1991 Apr;37(2):173-82
references.pmid: 1715499
references.citation: Nat Methods. 2007 Feb;4(2):127-9
references.pmid: 17259991
references.citation: Nat Protoc. 2009;4(10):1551-9
references.pmid: 19798087
references.citation: Nat Methods. 2008 Sep;5(9):805-11
references.pmid: 19160515
references.citation: Mol Ther. 2001 Feb;3(2):225-32
references.pmid: 11237679
references.citation: Brain Cell Biol. 2008 Aug;36(1-4):69-86
references.pmid: 18941901
references.citation: J Neurophysiol. 2007 May;97(5):3751-62
references.pmid: 17360827
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 1;106(48):20482-6
references.pmid: 19918085
references.citation: Nature. 1997 Aug 28;388(6645):882-7
references.pmid: 9278050
references.citation: J Neurosci. 2004 Oct 27;24(43):9598-611
references.pmid: 15509747
references.citation: Biophys J. 1996 Feb;70(2):1069-81
references.pmid: 8789126
references.citation: Nat Protoc. 2006;1(1):380-6
references.pmid: 17406260
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7319-24
references.pmid: 12777621
references.citation: J Neurosci Methods. 2005 Apr 30;143(2):151-8
references.pmid: 15814147
references.citation: J Neurophysiol. 2007 Sep;98(3):1791-805
references.pmid: 17634346
references.citation: Nat Neurosci. 1999 Nov;2(11):989-96
references.pmid: 10526338
references.citation: PLoS Biol. 2007 Jul;5(7):e189
references.pmid: 17622195
references.citation: PLoS One. 2008 Mar 19;3(3):e1796
references.pmid: 18350138