title: Stereotaxic injection of a viral vector for conditional gene manipulation in the mouse spinal cord.
abstract: Intraparenchymal injection of a viral vector enables conditional gene manipulation in distinct populations of neurons or particular regions of the central nervous system. We demonstrate a stereotaxic injection technique that allows targeted gene expression or silencing in the dorsal horn of the mouse spinal cord. The surgical procedure is brief. It requires laminectomy of a single vertebra, providing for quick recovery of the animal and unimpaired motility of the spine. Controlled injection of a small vector suspension volume at low speed and use of a microsyringe with beveled glass cannula minimize the tissue lesion. The local immune response to the vector depends on the intrinsic properties of the virus employed; in our experience, it is minor and short-lived when a recombinant adeno-associated virus is used. A reporter gene such as enhanced green fluorescent protein facilitates monitoring spatial distribution of the vector, and the efficacy and cellular specificity of the transfection.
authors.affiliation: Département Nociception et Douleur, Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique CNRS.
authors.forename: Perrine
authors.initials: P
authors.lastname: Inquimbert
authors.forename: Martin
authors.initials: M
authors.lastname: Moll
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Kohno
authors.forename: Joachim
authors.initials: J
authors.lastname: Scholz
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D003587:Cytomegalovirus / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007796:Laminectomy; D051379:Mice; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D020671:Posterior Horn Cells / Q000502:physiology / Q000821:virology; D011401:Promoter Regions, Genetic; D013116:Spinal Cord / Q000502:physiology* / Q000601:surgery / Q000821:virology; D013238:Stereotaxic Techniques
chemical_list: D049452:Green Fluorescent Proteins
references.citation: Hum Gene Ther. 2011 Sep;22(9):1129-35
references.pmid: 21443428
references.citation: Curr Opin Pharmacol. 2010 Oct;10(5):534-42
references.pmid: 20620113
references.citation: Nat Med. 2006 Jun;12(6):677-81
references.pmid: 16715092
references.citation: Nat Rev Genet. 2001 Oct;2(10):743-55
references.pmid: 11584291
references.citation: Nat Methods. 2011 Sep;8(9):745-52
references.pmid: 21985008
references.citation: Nat Rev Neurosci. 2010 Dec;11(12):823-36
references.pmid: 21068766
references.citation: J Neurosci. 2003 Jun 15;23(12):5031-40
references.pmid: 12832526
references.citation: J Clin Invest. 2010 Apr;120(4):1253-64
references.pmid: 20234094
references.citation: J Vis Exp. 2011 Jul 10;(53):e2834
references.pmid: 21775959
references.citation: Proc Natl Acad Sci U S A. 2007 Dec 11;104(50):20096-101
references.pmid: 18056815
references.citation: Mol Pain. 2009 Sep 08;5:52
references.pmid: 19737386
references.citation: J Neurosci. 2007 May 30;27(22):6068-78
references.pmid: 17537979
references.citation: Neuron. 2011 Dec 22;72(6):938-50
references.pmid: 22196330
references.citation: Neuron. 2008 Mar 13;57(5):634-60
references.pmid: 18341986
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 14;107(50):21848-53
references.pmid: 21115815
references.citation: Nat Genet. 1994 Oct;8(2):148-54
references.pmid: 7842013
references.citation: Nat Rev Neurosci. 2003 May;4(5):353-64
references.pmid: 12728263
references.citation: J Neurosci. 2006 Mar 15;26(11):2923-32
references.pmid: 16540569
references.citation: Hum Mol Genet. 2010 Jan 1;19(1):147-58
references.pmid: 19837699
references.citation: J Clin Invest. 2011 Aug;121(8):3029-41
references.pmid: 21785219