Targeting single neuronal networks for gene expression and cell labeling in vivo.
To understand fine-scale structure and function of single mammalian neuronal networks, we developed and validated a strategy to genetically target and trace monosynaptic inputs to a single neuron in vitro and in vivo. The strategy independently targets a neuron and its presynaptic network for specific gene expression and fine-scale labeling, using single-cell electroporation of DNA to target infection and monosynaptic retrograde spread of a genetically modifiable rabies virus. The technique is highly reliable, with transsynaptic labeling occurring in every electroporated neuron infected by the virus. Targeting single neocortical neuronal networks in vivo, we found clusters of both spiny and aspiny neurons surrounding the electroporated neuron in each case, in addition to intricately labeled distal cortical and subcortical inputs. This technique, broadly applicable for probing and manipulating single neuronal networks with single-cell resolution in vivo, may help shed new light on fundamental mechanisms underlying circuit development and information processing by neuronal networks throughout the brain.
Marshel
James H
JH
Systems Neurobiology Laboratories, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
Mori
Takuma
T
Nielsen
Kristina J
KJ
Callaway
Edward M
EM
D000818:Animals; D018274:Electroporation; D015870:Gene Expression; D005822:Genetic Vectors; D006652:Histological Techniques; D066298:In Vitro Techniques; D051379:Mice; D019579:Neocortex / Q000166:cytology* / Q000378:metabolism / Q000821:virology; D009434:Neural Pathways / Q000166:cytology / Q000502:physiology*; D056347:Neuroanatomical Tract-Tracing Techniques / Q000379:methods*; D056325:Neuronal Tract-Tracers; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology* / Q000821:virology; D017729:Presynaptic Terminals / Q000378:metabolism / Q000821:virology; D017966:Pyramidal Cells / Q000166:cytology / Q000502:physiology; D011820:Rabies virus / Q000235:genetics; D051381:Rats; D015203:Reproducibility of Results; D014793:Visual Cortex / Q000166:cytology / Q000378:metabolism / Q000821:virology
D056325:Neuronal Tract-Tracers
Mol Cell. 2005 Jan 7;17(1):93-102
15629720
Pflugers Arch. 2007 Jul;454(4):675-88
17334778
J Virol. 1993 Apr;67(4):1811-6
8383211
Curr Opin Neurobiol. 2009 Aug;19(4):415-21
19674891
Science. 2001 Aug 3;293(5531):868-72
11486089
Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):7310-4
8692989
Nat Neurosci. 2006 Dec;9(12):1472-3
17099707
Nature. 2009 Feb 26;457(7233):1142-5
19151697
Pflugers Arch. 2002 Feb;443(4):664-70
11907835
J Comp Neurol. 1991 Apr 8;306(2):332-43
1711059
Nature. 2007 Nov 1;450(7166):56-62
17972876
EMBO J. 1994 Sep 15;13(18):4195-203
7925265
Nat Protoc. 2009;4(6):862-9
19444243
Nat Neurosci. 2005 Nov;8(11):1552-9
16222228
Am J Anat. 1970 Apr;127(4):321-55
4985058
Neuron. 2009 Jan 29;61(2):301-16
19186171
J Virol. 2001 Oct;75(19):9339-44
11533197
J Comp Neurol. 1995 Jun 5;356(3):457-80
7642806
J Neurosci Methods. 2000 Nov 15;103(1):63-71
11074096
Cereb Cortex. 2003 Jan;13(1):5-14
12466210
PLoS Biol. 2005 Mar;3(3):e68
15737062
Synapse. 2006 Nov;60(6):437-40
16881073
Nat Protoc. 2006;1(1):241-5
17406239
Cereb Cortex. 2002 Sep;12(9):936-53
12183393
J Physiol. 1997 Apr 15;500 ( Pt 2):409-40
9147328
Trends Neurosci. 2005 Oct;28(10):541-51
16122815
J Gen Virol. 2000 Sep;81(Pt 9):2147-2153
10950970
Annu Rev Neurosci. 2004;27:419-51
15217339
J Neurosci. 2003 Sep 10;23(23):8432-44
12968006
Nat Methods. 2008 Jan;5(1):61-7
18157136
Neuron. 2003 Sep 11;39(6):911-8
12971892
Brain Res. 1993 Aug 13;619(1-2):146-56
7690671
Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3555-60
19221032
J Neurosci. 1998 Jun 1;18(11):4145-54
9592094
J Neurosci Methods. 2003 Jun 15;126(1):91-8
12788505
J Neurocytol. 2002 Mar-Jun;31(3-5):277-87
12815247
Sci Am. 1979 Sep;241(3):219-32
115087
Neuron. 2001 Mar;29(3):583-91
11301019
Nature. 2005 Feb 10;433(7026):597-603
15660108
Nat Med. 2008 Oct;14(10):1123-7
18836461
Nature. 2009 Feb 26;457(7233):1133-6
19151698
Nat Protoc. 2010 Mar;5(3):595-606
20203674
Nat Methods. 2007 Jan;4(1):47-9
17179932
J Neurosci. 2006 Dec 6;26(49):12789-98
17151282
J Physiol. 2002 Feb 1;538(Pt 3):803-22
11826166
Eur J Neurosci. 2004 Nov;20(10):2681-90
15548211
Nat Neurosci. 2007 May;10(5):663-8
17435752
Brain Res Brain Res Rev. 1998 May;26(2-3):113-35
9651498
Front Neurosci. 2007 Oct 15;1(1):19-42
18982117
Prog Neurobiol. 1992 Dec;39(6):563-607
1410442
Trends Neurosci. 2005 Jul;28(7):387-94
15935485
Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3619-24
10737806
Nature. 2008 Aug 14;454(7206):881-5
18633351
J Neurosci. 1999 Apr 15;19(8):2876-86
10191305
J Comp Neurol. 1989 Aug 22;286(4):442-55
2778101
Cell. 1996 Mar 22;84(6):941-51
8601317
J Neurophysiol. 1993 Dec;70(6):2354-69
8120587
Nat Neurosci. 2006 Apr;9(4):534-42
16547512
Neuron. 2007 Mar 1;53(5):639-47
17329205
Neuron. 2008 Mar 13;57(5):634-60
18341986
Science. 2003 Dec 12;302(5652):1981-4
14671309
Annu Rev Genet. 2002;36:153-73
12429690
Nature. 2005 Feb 24;433(7028):868-73
15729343