Generation of Cre-transgenic mice using Dlx1/Dlx2 enhancers and their characterization in GABAergic interneurons

Gregory B. Potter; Magdalena A. Petryniak; Eugenia Shevchenko; Gabriel L. McKinsey; Marc Ekker; John L.R. Rubenstein

doi:10.1016/j.mcn.2008.10.003

Generation of Cre-transgenic mice using Dlx1/Dlx2 enhancers and their characterization in GABAergic interneurons

Gregory B. Potter
, Magdalena A. Petryniak
, Eugenia Shevchenko
, Gabriel L. McKinsey
, Marc Ekker
, John L.R. Rubenstein

Research output: Contribution to journal › Article › peer-review

99 Scopus citations

Abstract

DLX1 and DLX2 transcription factors are necessary for forebrain GABAergic neuron differentiation, migration, and survival. We generated transgenic mice that express Cre-recombinase under the control of two ultra-conserved DNA elements near the Dlx1 and 2 locus termed I12b and URE2. We show that Cre-recombinase is active in a "Dlx-pattern" in the embryonic forebrain of transgenic mice. I12b-Cre is more active than URE2-Cre in the medial ganglionic eminences and its derivatives. Fate-mapping of EGFP+ cells in adult Cre;Z/EG animals demonstrated that GABAergic neurons, but not glia, are labeled. Most NPY+, nNOS+, parvalbumin+, and somatostatin+ cells are marked by I12b-Cre in the cortex and hippocampus, while 25-40% of these interneuron subtypes are labeled by URE2-Cre. Labeling of neurons generated between E12.5 to E15.5 indicated differences in birth-dates of EGFP+ cells that populate the olfactory bulb, hippocampus, and cortex. Finally, we provide the first in vivo evidence that both I12b and URE2 are direct targets of DLX2 and require Dlx1 and Dlx2 expression for proper activity.

Original language	English (US)
Pages (from-to)	167-186
Number of pages	20
Journal	Molecular and Cellular Neuroscience
Volume	40
Issue number	2
DOIs	https://doi.org/10.1016/j.mcn.2008.10.003
State	Published - Feb 2009
Externally published	Yes

Funding

UCSF Comprehensive Cancer Center Transgenic Core performed the oocyte injections and generated the I12b-Cre and URE2-Cre transgenic mice. Louis Reichardt, Ben Cheyette and the Nikon Imaging Center at UCSF provided access to microscopes used in this study. We would like to thank M. Wegner for the SOX10 antibody, K. Yoshikawa for DLX2 antibody, and C. Stiles for OLIG1 and OLIG2 antibodies. We thank N. Ghanem for providing data prior to publication. G.B.P. made the I12b-Cre and URE2-Cre transgenes, characterized the different lines of I12b-Cre and URE2-Cre transgenic mice, maintained the mouse colonies, designed and implemented the experiments, photographed and quantified brain sections, performed statistical analysis on the data, wrote the manuscript, and prepared all tables and figures. M.A.P. helped with experimental design, interpreted the data, and prepared, sectioned, immunolabeled and photographed brain tissue. E.S. genotyped and maintained transgenic mice, sectioned, immunolabeled and photographed tissue, and quantified the birth-date data. G.L.M. performed the ChIP experiments. M.E. provided plasmids containing I12b and URE2 DNA, feedback on transgene construction, and comments on the manuscript. J.L.R. provided mentorship on experimental design, analysis, and manuscript preparation. G.B.P. was supported by the California Institute of Regenerative Medicine's Postdoctoral Fellowship; M.A.P. was funded in part by NICHD HD-07162H; M.E. was supported by CIHR grant #MOP-14460; JLR was supported by Nina Ireland, NIMH RO1 MH49428 and K05 MH065670.

Funders	Funder number
Nina Ireland
National Institutes of Health National Institute of Mental Health	RO1 MH49428, K05 MH065670
The National Institute of Child Health and Human Development	T32HD007162
California Institute for Regenerative Medicine
Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health
Canadian Institutes of Health Research	-14460

ASJC Scopus subject areas

Molecular Biology
Cellular and Molecular Neuroscience
Cell Biology

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10.1016/j.mcn.2008.10.003

Cite this

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title = "Generation of Cre-transgenic mice using Dlx1/Dlx2 enhancers and their characterization in GABAergic interneurons",

abstract = "DLX1 and DLX2 transcription factors are necessary for forebrain GABAergic neuron differentiation, migration, and survival. We generated transgenic mice that express Cre-recombinase under the control of two ultra-conserved DNA elements near the Dlx1 and 2 locus termed I12b and URE2. We show that Cre-recombinase is active in a {"}Dlx-pattern{"} in the embryonic forebrain of transgenic mice. I12b-Cre is more active than URE2-Cre in the medial ganglionic eminences and its derivatives. Fate-mapping of EGFP+ cells in adult Cre;Z/EG animals demonstrated that GABAergic neurons, but not glia, are labeled. Most NPY+, nNOS+, parvalbumin+, and somatostatin+ cells are marked by I12b-Cre in the cortex and hippocampus, while 25-40\% of these interneuron subtypes are labeled by URE2-Cre. Labeling of neurons generated between E12.5 to E15.5 indicated differences in birth-dates of EGFP+ cells that populate the olfactory bulb, hippocampus, and cortex. Finally, we provide the first in vivo evidence that both I12b and URE2 are direct targets of DLX2 and require Dlx1 and Dlx2 expression for proper activity.",

author = "Potter, \{Gregory B.\} and Petryniak, \{Magdalena A.\} and Eugenia Shevchenko and McKinsey, \{Gabriel L.\} and Marc Ekker and Rubenstein, \{John L.R.\}",

note = "Funding Information: UCSF Comprehensive Cancer Center Transgenic Core performed the oocyte injections and generated the I12b-Cre and URE2-Cre transgenic mice. Louis Reichardt, Ben Cheyette and the Nikon Imaging Center at UCSF provided access to microscopes used in this study. We would like to thank M. Wegner for the SOX10 antibody, K. Yoshikawa for DLX2 antibody, and C. Stiles for OLIG1 and OLIG2 antibodies. We thank N. Ghanem for providing data prior to publication. G.B.P. made the I12b-Cre and URE2-Cre transgenes, characterized the different lines of I12b-Cre and URE2-Cre transgenic mice, maintained the mouse colonies, designed and implemented the experiments, photographed and quantified brain sections, performed statistical analysis on the data, wrote the manuscript, and prepared all tables and figures. M.A.P. helped with experimental design, interpreted the data, and prepared, sectioned, immunolabeled and photographed brain tissue. E.S. genotyped and maintained transgenic mice, sectioned, immunolabeled and photographed tissue, and quantified the birth-date data. G.L.M. performed the ChIP experiments. M.E. provided plasmids containing I12b and URE2 DNA, feedback on transgene construction, and comments on the manuscript. J.L.R. provided mentorship on experimental design, analysis, and manuscript preparation. G.B.P. was supported by the California Institute of Regenerative Medicine's Postdoctoral Fellowship; M.A.P. was funded in part by NICHD HD-07162H; M.E. was supported by CIHR grant \#MOP-14460; JLR was supported by Nina Ireland, NIMH RO1 MH49428 and K05 MH065670. ",

year = "2009",

month = feb,

doi = "10.1016/j.mcn.2008.10.003",

language = "English (US)",

volume = "40",

pages = "167--186",

journal = "Molecular and Cellular Neuroscience",

issn = "1044-7431",

publisher = "Academic Press Inc.",

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AU - Petryniak, Magdalena A.

AU - Shevchenko, Eugenia

AU - McKinsey, Gabriel L.

AU - Ekker, Marc

AU - Rubenstein, John L.R.

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N2 - DLX1 and DLX2 transcription factors are necessary for forebrain GABAergic neuron differentiation, migration, and survival. We generated transgenic mice that express Cre-recombinase under the control of two ultra-conserved DNA elements near the Dlx1 and 2 locus termed I12b and URE2. We show that Cre-recombinase is active in a "Dlx-pattern" in the embryonic forebrain of transgenic mice. I12b-Cre is more active than URE2-Cre in the medial ganglionic eminences and its derivatives. Fate-mapping of EGFP+ cells in adult Cre;Z/EG animals demonstrated that GABAergic neurons, but not glia, are labeled. Most NPY+, nNOS+, parvalbumin+, and somatostatin+ cells are marked by I12b-Cre in the cortex and hippocampus, while 25-40% of these interneuron subtypes are labeled by URE2-Cre. Labeling of neurons generated between E12.5 to E15.5 indicated differences in birth-dates of EGFP+ cells that populate the olfactory bulb, hippocampus, and cortex. Finally, we provide the first in vivo evidence that both I12b and URE2 are direct targets of DLX2 and require Dlx1 and Dlx2 expression for proper activity.

AB - DLX1 and DLX2 transcription factors are necessary for forebrain GABAergic neuron differentiation, migration, and survival. We generated transgenic mice that express Cre-recombinase under the control of two ultra-conserved DNA elements near the Dlx1 and 2 locus termed I12b and URE2. We show that Cre-recombinase is active in a "Dlx-pattern" in the embryonic forebrain of transgenic mice. I12b-Cre is more active than URE2-Cre in the medial ganglionic eminences and its derivatives. Fate-mapping of EGFP+ cells in adult Cre;Z/EG animals demonstrated that GABAergic neurons, but not glia, are labeled. Most NPY+, nNOS+, parvalbumin+, and somatostatin+ cells are marked by I12b-Cre in the cortex and hippocampus, while 25-40% of these interneuron subtypes are labeled by URE2-Cre. Labeling of neurons generated between E12.5 to E15.5 indicated differences in birth-dates of EGFP+ cells that populate the olfactory bulb, hippocampus, and cortex. Finally, we provide the first in vivo evidence that both I12b and URE2 are direct targets of DLX2 and require Dlx1 and Dlx2 expression for proper activity.

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Generation of Cre-transgenic mice using Dlx1/Dlx2 enhancers and their characterization in GABAergic interneurons

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