Characterization of a mouse strain expressing Cre recombinase from the 3′ untranslated region of the dopamine transporter locus

Dopamine (DA) neurotransmission has been implicated in several neurological and psychiatric disorders. The dopamine transporter (DAT) is highly expressed in dopaminergic neurons of the ventral mesencephalon and regulates neurotransmission by transporting DA back into the presynaptic terminals. To mediate restricted DNA recombination events into DA neurons using the Cre/loxP technology, we have generated a knockin mouse expressing Cre recombinase under the transcriptional control of the endogenous DAT promoter. To minimize interference with DAT function by preservation of both DAT alleles, Cre recombinase expression was driven from the 3′ untranslated region (3′UTR) of the endogenous DAT gene by means of an internal ribosomal entry sequence. Crossing this murine line with a LacZ reporter showed colocalization of DAT immunocytochemistry and β‐galactosidase staining in all regions analyzed. This knockin mouse can be used for generating tissue specific knockouts in mice carrying genes flanked by loxP sites, and will facilitate the analysis of gene function in dopaminergic neurons. genesis 44:383–390, 2006. Published 2006 Wiley‐Liss, Inc.

[1]  T. Jessell,et al.  Genetic ablation reveals that the roof plate is essential for dorsal interneuron specification , 2000, Nature.

[2]  H. Bengtsson,et al.  Transgenic expression of Cre recombinase from the tyrosine hydroxylase locus , 2004, Genesis.

[3]  F. Gonon,et al.  Increased rewarding properties of morphine in dopamine‐transporter knockout mice , 2000, The European journal of neuroscience.

[4]  Béatrice Conne,et al.  The 3′ untranslated region of messenger RNA: A molecular ‘hotspot’ for pathology? , 2000, Nature Medicine.

[5]  E. Nestler Is there a common molecular pathway for addiction? , 2005, Nature Neuroscience.

[6]  Eric Mick,et al.  Genetics of attention deficit hyperactivity disorder. , 1994, Child and adolescent psychiatric clinics of North America.

[7]  C. Lobe,et al.  Z/AP, a double reporter for cre-mediated recombination. , 1999, Developmental biology.

[8]  M. Kreek,et al.  Quantitation of dopamine transporter mrna in the rat brain: Mapping, effects of “binge” cocaine administration and withdrawal , 1997, Synapse.

[9]  René Hen,et al.  Targeted gene expression in dopamine and serotonin neurons of the mouse brain , 2005, Journal of Neuroscience Methods.

[10]  Philippe Soriano Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[11]  K. Fuxe,et al.  Demonstration and mapping of central neurons containing dopamine, noradrenaline, and 5-hydroxytryptamine and their reactions to psychopharmaca. , 1966, Pharmacological reviews.

[12]  T. Arinami,et al.  Identification of a novel polymorphism of the human dopamine transporter (DAT1) gene and the significant association with alcoholism , 1999, Molecular Psychiatry.

[13]  M. Caron,et al.  Decreased ethanol preference and consumption in dopamine transporter female knock-out mice. , 2002, Alcoholism, clinical and experimental research.

[14]  M. Smidt,et al.  Developmental origin and fate of meso-diencephalic dopamine neurons , 2006, Progress in Neurobiology.

[15]  P. McKenna,et al.  Co-expression of dopamine transporter mRNA and tyrosine hydroxylase mRNA in ventral mesencephalic neurones. , 1993, Brain research. Molecular brain research.

[16]  Luis Puelles,et al.  Cortical Excitatory Neurons and Glia, But Not GABAergic Neurons, Are Produced in the Emx1-Expressing Lineage , 2002, The Journal of Neuroscience.

[17]  E. Stanley,et al.  Efficient Cre-mediated deletion in cardiac progenitor cells conferred by a 3'UTR-ires-Cre allele of the homeobox gene Nkx2-5. , 2002, The International journal of developmental biology.

[18]  Andras Nagy,et al.  Cre recombinase: The universal reagent for genome tailoring , 2000, Genesis.

[19]  M. Kozak How strong is the case for regulation of the initiation step of translation by elements at the 3' end of eukaryotic mRNAs? , 2004, Gene.

[20]  R. Mark Wightman,et al.  Hyperlocomotion and indifference to cocaine and amphetamine in mice lacking the dopamine transporter , 1996, Nature.

[21]  I. Sora,et al.  Sex-Dependent Modulation of Ethanol Consumption in Vesicular Monoamine Transporter 2 (VMAT2) and Dopamine Transporter (DAT) Knockout Mice , 2003, Neuropsychopharmacology.

[22]  B. Madras,et al.  Polymorphisms in the 3′-untranslated region of human and monkey dopamine transporter genes affect reporter gene expression , 2002, Molecular Psychiatry.

[23]  S. Amara,et al.  Cell-type-specific expression of catecholamine transporters in the rat brain , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[24]  B. Hoffer,et al.  Glial cell line-derived neurotrophic factor receptor α1 availability regulates glial cell line-derived neurotrophic factor signaling: evidence from mice carrying one or two mutated alleles , 1999, Neuroscience.

[25]  Michael J Owen,et al.  The genetics of attention deficit hyperactivity disorder. , 2005, Human molecular genetics.

[26]  H. Meijer,et al.  Mechanisms of translational control by the 3' UTR in development and differentiation. , 2005, Seminars in cell & developmental biology.

[27]  R. Wightman,et al.  Profound neuronal plasticity in response to inactivation of the dopamine transporter. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[28]  K. Fuxe,et al.  Localization of monoamines in the lower brain stem , 1964, Experientia.

[29]  David A Lewis,et al.  Catching Up on Schizophrenia Natural History and Neurobiology , 2000, Neuron.

[30]  P. Mountford,et al.  Internal ribosome entry sites and dicistronic RNAs in mammalian transgenesis , 1995, Trends in Genetics.