title: Chronic cellular imaging of mouse visual cortex during operant behavior and passive viewing.
abstract: Nearby neurons in mammalian neocortex demonstrate a great diversity of cell types and connectivity patterns. The importance of this diversity for computation is not understood. While extracellular recording studies in visual cortex have provided a particularly rich description of behavioral modulation of neural activity, new methods are needed to dissect the contribution of specific circuit elements in guiding visual perception. Here, we describe a method for three-dimensional cellular imaging of neural activity in the awake mouse visual cortex during active discrimination and passive viewing of visual stimuli. Head-fixed mice demonstrated robust discrimination for many hundred trials per day after initial task acquisition. To record from multiple neurons during operant behavior with single-trial resolution and minimal artifacts, we built a sensitive microscope for two-photon calcium imaging, capable of rapid tracking of neurons in three dimensions. We demonstrate stable recordings of cellular calcium activity during discrimination behavior across hours, days, and weeks, using both synthetic and genetically encoded calcium indicators. When combined with molecular and genetic technologies in mice (e.g., cell-type specific transgenic labeling), this approach allows the identification of neuronal classes in vivo. Physiological measurements from distinct classes of neighboring neurons will enrich our understanding of the coordinated roles of diverse elements of cortical microcircuits in guiding sensory perception and perceptual learning. Further, our method provides a high-throughput, chronic in vivo assay of behavioral influences on cellular activity that is applicable to a wide range of mouse models of neurologic disease.
authors.affiliation: Department of Neurobiology, Harvard Medical School Boston, MA, USA.
authors.forename: Mark L
authors.initials: ML
authors.lastname: Andermann
authors.forename: A M
authors.initials: AM
authors.lastname: Kerlin
authors.forename: R C
authors.initials: RC
authors.lastname: Reid
references.citation: Nat Methods. 2004 Oct;1(1):31-7
references.pmid: 15782150
references.citation: Neuron. 2007 Jul 5;55(1):131-41
references.pmid: 17610822
references.citation: Nat Methods. 2007 Jan;4(1):73-9
references.pmid: 17143280
references.citation: Exp Neurol. 2006 Feb;197(2):291-300
references.pmid: 16039651
references.citation: J Neurosci. 1996 Nov 15;16(22):7376-89
references.pmid: 8929444
references.citation: Nature. 2009 Oct 15;461(7266):923-9
references.pmid: 19829372
references.citation: Neuron. 2007 Oct 4;56(1):43-57
references.pmid: 17920014
references.citation: Nat Neurosci. 2008 Aug;11(8):974-82
references.pmid: 18604204
references.citation: J Neurophysiol. 1987 Apr;57(4):977-1001
references.pmid: 3585466
references.citation: J Neurophysiol. 2006 Nov;96(5):2377-87
references.pmid: 16855106
references.citation: Nature. 2009 Oct 15;461(7266):941-6
references.pmid: 19829374
references.citation: Nat Methods. 2008 Sep;5(9):797-804
references.pmid: 19160514
references.citation: J Comp Neurol. 2003 Dec 1;467(1):60-79
references.pmid: 14574680
references.citation: J Neurosci. 2007 Feb 21;27(8):2145-9
references.pmid: 17314309
references.citation: Somatosens Mot Res. 2001;18(3):211-22
references.pmid: 11562084
references.citation: Curr Opin Neurobiol. 2009 Oct;19(5):520-9
references.pmid: 19854041
references.citation: Rev Neurosci. 2002;13(4):287-98
references.pmid: 12542258
references.citation: Neuron. 2009 Sep 24;63(6):747-60
references.pmid: 19778505
references.citation: Exp Brain Res. 1991;83(2):340-8
references.pmid: 2022243
references.citation: J Neurosci Methods. 2009 Jan 15;176(1):1-15
references.pmid: 18789968
references.citation: J Neurosci. 2007 Jul 25;27(30):8122-37
references.pmid: 17652603
references.citation: J Comp Neurol. 1992 Jun 1;320(1):62-8
references.pmid: 1401242
references.citation: Nat Methods. 2009 Dec;6(12):875-81
references.pmid: 19898485
references.citation: J Neurosci. 2008 Jul 16;28(29):7399-411
references.pmid: 18632944
references.citation: Biomed Eng Online. 2003 May 17;2:13
references.pmid: 12801419
references.citation: Neuron. 2007 Jun 7;54(5):677-96
references.pmid: 17553419
references.citation: J Neurosci. 2005 Nov 23;25(47):11023-33
references.pmid: 16306415
references.citation: Neuron. 2000 Sep;27(3):461-8
references.pmid: 11055429
references.citation: Annu Rev Neurosci. 1995;18:193-222
references.pmid: 7605061
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):14063-8
references.pmid: 16157876
references.citation: Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10554-9
references.pmid: 15247428
references.citation: Nat Neurosci. 2008 Jul;11(7):749-51
references.pmid: 18552841
references.citation: J Neurosci Methods. 2000 Jun 30;99(1-2):101-10
references.pmid: 10936649
references.citation: Proc Natl Acad Sci U S A. 2008 May 27;105(21):7588-93
references.pmid: 18508976
references.citation: J Neurophysiol. 2000 Mar;83(3):1677-92
references.pmid: 10712488
references.citation: Eur J Nucl Med Mol Imaging. 2008 Mar;35 Suppl 1:S99-106
references.pmid: 18193219
references.citation: Neuron. 2002 Jan 31;33(3):425-37
references.pmid: 11832229
references.citation: Curr Opin Neurobiol. 2009 Aug;19(4):415-21
references.pmid: 19674891
references.citation: Curr Biol. 2004 May 4;14(9):744-51
references.pmid: 15120065
references.citation: Vision Res. 2004 Dec;44(28):3411-8
references.pmid: 15536009
references.citation: J Neurophysiol. 2008 May;99(5):2158-82
references.pmid: 18287553
references.citation: Nat Methods. 2008 Sep;5(9):805-11
references.pmid: 19160515
references.citation: Nature. 2008 Aug 28;454(7208):1110-4
references.pmid: 18633352
references.citation: J Neurosci. 1999 Jan 1;19(1):431-41
references.pmid: 9870971
references.citation: Behav Brain Res. 1987 Apr;24(1):81-4
references.pmid: 3580117
references.citation: J Comp Neurol. 1975 Apr 1;160(3):269-90
references.pmid: 1112925
references.citation: Neuron. 2009 Aug 27;63(4):508-22
references.pmid: 19709632
references.citation: Trends Neurosci. 2006 Mar;29(3):160-6
references.pmid: 16443289
references.citation: Nature. 2005 Feb 10;433(7026):597-603
references.pmid: 15660108
references.citation: J Comp Neurol. 2007 May 20;502(3):339-57
references.pmid: 17366604
references.citation: Cereb Cortex. 2000 Apr;10(4):359-70
references.pmid: 10769248
references.citation: J Neurosci. 2009 Nov 4;29(44):13751-60
references.pmid: 19889987
references.citation: Science. 2009 Feb 27;323(5918):1211-5
references.pmid: 19251629
references.citation: J Physiol. 2003 Aug 15;551(Pt 1):13-32
references.pmid: 12813159
references.citation: Nat Neurosci. 2002 Nov;5(11):1203-9
references.pmid: 12379861
references.citation: Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19557-62
references.pmid: 19889973
references.citation: Nat Protoc. 2006;1(2):936-46
references.pmid: 17406327
references.citation: J Neurosci. 2010 Feb 3;30(5):1947-67
references.pmid: 20130203
references.citation: Nat Methods. 2008 Nov;5(11):935-8
references.pmid: 18836457
references.citation: J Neurosci. 2004 Oct 27;24(43):9572-9
references.pmid: 15509744
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7319-24
references.pmid: 12777621
references.citation: Neuron. 2006 Aug 3;51(3):339-49
references.pmid: 16880128
references.citation: Nat Neurosci. 2006 Oct;9(10):1240-2
references.pmid: 16964255
references.citation: Genes Brain Behav. 2006 Jul;5(5):389-403
references.pmid: 16879633
references.citation: Nat Methods. 2009 Dec;6(12):883-9
references.pmid: 19898484
references.citation: Neuron. 2007 Jun 21;54(6):961-72
references.pmid: 17582335
references.citation: Neurosci Res. 2004 May;49(1):123-31
references.pmid: 15099710
references.citation: Neuron. 2010 Feb 25;65(4):472-9
references.pmid: 20188652
references.citation: Methods. 1999 Jun;18(2):215-21
references.pmid: 10356353
references.citation: Nat Neurosci. 2007 May;10(5):631-9
references.pmid: 17450136
references.citation: J Neurosci. 1999 May 15;19(10):4046-64
references.pmid: 10234033
references.citation: Nat Neurosci. 2006 May;9(5):608-10
references.pmid: 16617340
references.citation: J Neurosci. 2008 Jul 23;28(30):7520-36
references.pmid: 18650330
references.citation: Opt Express. 2008 Apr 14;16(8):5556-64
references.pmid: 18542658
references.citation: Nature. 2008 Aug 14;454(7206):881-5
references.pmid: 18633351
references.citation: J Physiol. 2003 Aug 15;551(Pt 1):67-78
references.pmid: 12844507
references.citation: Pflugers Arch. 2002 Jul;444(4):491-8
references.pmid: 12136268
references.citation: J Neurophysiol. 1983 Dec;50(6):1415-32
references.pmid: 6663335
references.citation: Neuron. 2008 Jul 31;59(2):214-25
references.pmid: 18667150
references.citation: J Comp Neurol. 1980 Sep 1;193(1):203-22
references.pmid: 6776165
references.citation: Exp Brain Res. 1988;69(3):594-612
references.pmid: 3371440
references.citation: Neurobiol Learn Mem. 2006 Mar;85(2):164-72
references.pmid: 16290194
references.citation: PLoS Biol. 2007 Jul;5(7):e189
references.pmid: 17622195
references.citation: Neuron. 2006 May 18;50(4):617-29
references.pmid: 16701211
references.citation: J Neurosci. 2009 Aug 26;29(34):10520-32
references.pmid: 19710305