tailieunhanh - Báo cáo khoa học: Analysis of dopamine transporter gene expression pattern ) generation of DAT-iCre transgenic mice

Analysis of dopamine transporter gene expression pattern ) generation of DAT-iCre transgenic mice ´ Marc Turiault1,*, Sebastien Parnaudeau1,*, Aude Milet1, Rosanna Parlato2, Jean-Denis Rouzeau1, Monique Lazar1 and Francois Tronche1 ` 1 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, College de France, Institut de Biologie, Paris, France 2 Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany Keywords Cre ⁄ loxP system; dopaminergic cells; dopamine transporter; gene expression; transgenic mice Correspondence F. Tronche, UMR7148 CNRS, 11 place Marcelin Berthelot, 75005 Paris, France Fax: +33 1 44 27 13 22 Tel: +33 1 44 27 13 08 E-mail: *These authors. | ễFEBS Journal Analysis of dopamine transporter gene expression pattern - generation of DAT-iCre transgenic mice Marc Turiault1 Sébastien Parnaudeau1 Aude Milet1 Rosanna Parlato2 Jean-Denis Rouzeau1 Monique Lazar1 and Francois Tronche1 1 CNRS UMR7148 Molecular Genetics Neurophysiology and Behavior College de France Institut de Biologie Paris France 2 Molecular Biology of the Cell I German Cancer Research Center Heidelberg Germany Keywords Cre loxP system dopaminergic cells dopamine transporter gene expression transgenic mice Correspondence F. Tronche UMR7148 CNRS 11 place Marcelin Berthelot 75005 Paris France Fax 33 1 44 27 13 22 Tel 33 1 44 27 13 08 E-mail These authors contributed equally to this work Received 16 February 2007 revised 10 May 2007 accepted 16 May 2007 doi The dopamine transporter is an essential component of the dopaminergic synapse. It is located in the presynaptic neurons and regulates extracellular dopamine levels. We generated a transgenic mouse line expressing the Cre recombinase under the control of the regulatory elements of the dopamine transporter gene for investigations of gene function in dopaminergic neurons. The codon-improved Cre recombinase iCre gene was inserted into the dopamine transporter gene on a bacterial artificial chromosome. The pattern of expression of the bacterial artificial chromosome-dopamine transporter-iCre transgene was similar to that of the endogenous dopamine transporter gene as shown by immunohistochemistry. Recombinase activity was further studied in mice carrying both the bacterial artificial chromo-some-dopamine transporter-iCre transgene and a construct expressing the b-galactosidase gene after Cre-mediated recombination. In situ studies showed that b-galactosidase 5-bromo-4-chloroindol-3-yl b-D-galactoside staining and the dopamine transporter immunofluorescence had identical distributions in the ventral midbrain. We used this animal model to study

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