title: Simultaneous expression of different transgenes in neurons and glia by combining in utero electroporation with the Tol2 transposon-mediated gene transfer system.
abstract: In utero electroporation is widely used to study neuronal development and function by introducing plasmid DNA into neural progenitors during embryogenesis. This is an effective and convenient method of introducing plasmid DNA into neural precursors and is suitable for manipulating gene expression in cells of the CNS. However, the applicability of this technique is comparatively limited to neuronal research, as the plasmid DNA introduced into neural progenitors during embryogenesis is diluted by cell proliferation and is not stably maintained in glial cells generated around and after birth. To overcome this limitation, we applied the Tol2 transposon system, which integrates a transgene into the genome of the host cell, to in utero electroporation. With this system, we confirmed that the transgene was effectively maintained in the progeny of embryonic neural precursors, astrocytes and oligodendrocytes. Using the glial promoters GFAP and S100beta, targeted and stable expressions of transgenes in glia were obtained, which enabled the expression of different transgenes simultaneously in neurons and glia. Glia-targeted expression of the transgene that causes neuronal migration defect was achieved without the defect. Thus, use of the Tol2 transposon system in combination with in utero electroporation is a powerful method for studying glia-neuron interactions in vivo.
authors.affiliation: Department of Genetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Yoshida
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: Keiko
authors.initials: K
authors.lastname: Nonomura
authors.forename: Koichi
authors.initials: K
authors.lastname: Kawakami
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Miura
mesh_terms: D000818:Animals; D004251:DNA Transposable Elements; D018274:Electroporation / Q000379:methods*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D019076:Transgenes; D019895:Transposases / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D005904:Glial Fibrillary Acidic Protein; D009414:Nerve Growth Factors; D011993:Recombinant Fusion Proteins; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D019895:Transposases
references.citation: Nature. 2001 Feb 8;409(6821):714-20
references.pmid: 11217860
references.citation: EMBO J. 2003 Aug 15;22(16):4190-201
references.pmid: 12912917
references.citation: Dev Biol. 2004 Jun 1;270(1):200-13
references.pmid: 15136150
references.citation: Nature. 2004 Apr 15;428(6984):767-71
references.pmid: 15085137
references.citation: Science. 1988 Sep 9;241(4871):1342-5
references.pmid: 3137660
references.citation: Neuron. 1993 Feb;10(2):201-12
references.pmid: 8439409
references.citation: Eur J Neurosci. 2006 Oct;24(7):1897-906
references.pmid: 17067293
references.citation: J Neurochem. 1990 Oct;55(4):1180-8
references.pmid: 2398353
references.citation: Dev Dyn. 2007 May;236(5):1273-86
references.pmid: 17377980
references.citation: Dev Biol. 2008 Aug 15;320(2):456-68
references.pmid: 18582453
references.citation: Neuroscience. 2001;103(4):865-72
references.pmid: 11301197
references.citation: Dev Growth Differ. 1999 Feb;41(1):59-72
references.pmid: 10445503
references.citation: Dev Biol. 2007 May 15;305(2):625-36
references.pmid: 17359965
references.citation: Genesis. 2006 Sep;44(9):438-45
references.pmid: 16906529
references.citation: Glia. 2009 Aug 1;57(10):1115-29
references.pmid: 19115384
references.citation: J Biol Chem. 2005 Feb 11;280(6):5082-8
references.pmid: 15557338
references.citation: Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11403-8
references.pmid: 11027340
references.citation: Neurochem Int. 2004 Jul-Aug;45(2-3):191-202
references.pmid: 15145537
references.citation: J Neurosci. 2006 Jan 18;26(3):1045-56
references.pmid: 16421324
references.citation: Dev Biol. 2001 Dec 1;240(1):237-46
references.pmid: 11784059
references.citation: Cereb Cortex. 2009 Jul;19 Suppl 1:i120-5
references.pmid: 19395528
references.citation: Trends Neurosci. 2009 Jun;32(6):303-11
references.pmid: 19398132
references.citation: Cell. 2008 Nov 14;135(4):738-48
references.pmid: 19013281
references.citation: J Comp Neurol. 2003 Mar 17;457(4):404-19
references.pmid: 12561079
references.citation: Development. 2006 Oct;133(20):4085-96
references.pmid: 16987869
references.citation: Cell. 2008 Nov 14;135(4):749-62
references.pmid: 19013282
references.citation: Prog Neurobiol. 2008 Dec 11;86(4):342-67
references.pmid: 18948166
references.citation: Neuron. 2007 May 3;54(3):357-69
references.pmid: 17481390
references.citation: Dev Biol. 2007 May 15;305(2):616-24
references.pmid: 17362912
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3581-6
references.pmid: 18299565
references.citation: Science. 2001 Nov 2;294(5544):1071-4
references.pmid: 11567107
references.citation: J Neurosci. 1999 Dec 1;19(23):10357-71
references.pmid: 10575033
references.citation: J Neurosci Res. 2007 Dec;85(16):3567-73
references.pmid: 17671991
references.citation: Genetics. 2004 Feb;166(2):895-9
references.pmid: 15020474
references.citation: Biochem Biophys Res Commun. 1997 Jan 13;230(2):376-80
references.pmid: 9016787
references.citation: Gene. 2008 Dec 1;425(1-2):64-8
references.pmid: 18775483