Abstract
BackgroundDopamine-activated signaling regulates locomotor and emotional responses and alterations in dopamine-signaling are responsible of several psychomotor disorders. In order to identify specific functions of these pathways, the Cre/loxP system has been used. Here, we describe the generation and the characterization of a transgenic mouse line expressing the Cre recombinase in dopaminoceptive neurons. To this purpose, we used as expression vector a 140 kb yeast artificial chromosome (YAC) containing the dopamine D1 receptor gene (Drd1a).ResultsIn the chosen line, D1Cre, the spatio-temporal pattern of Cre expression closely recapitulated that of the endogenous Drd1a gene, as assessed by immunohistological approaches in embryonic and adult stages. Efficiency of recombination was confirmed by crossing D1Cre with three different loxP lines (Creb1loxP, CaMKIVloxP and GRloxP) and with the R26R reporter. In the three loxP lines studied, recombination was restricted to the area of Cre expression.ConclusionIn view of the patterns of recombination restricted to the major dopaminoceptive regions as seen in the context of the CREB, CaMKIV and GR mutations, the D1Cre line will be a useful tool to dissect the contributions of specific genes to biological processes involving dopamine signaling.
Highlights
Dopamine-activated signaling regulates locomotor and emotional responses and alterations in dopamine-signaling are responsible of several psychomotor disorders
Detailed analysis of the Cre expression pattern was conducted with line A which carries three copies of the yeast artificial chromosome (YAC) transgene
In order to verify that Cre is expressed in neurons expressing the D1R mRNA, we have performed in situ hybridization with a specific D1R riboprobe in combination with immunohistochemical detection of Cre expression in striatum and cortex (Fig. 3a, b)
Summary
Dopamine-activated signaling regulates locomotor and emotional responses and alterations in dopamine-signaling are responsible of several psychomotor disorders. We describe the generation and the characterization of a transgenic mouse line expressing the Cre recombinase in dopaminoceptive neurons. To this purpose, we used as expression vector a 140 kb yeast artificial chromosome (YAC) containing the dopamine D1 receptor gene (Drd1a). Dopaminergic neurons are few (e.g. 10–20,000 in the rat brain), they regulate a number of physiological, behavioral and cognitive functions, including regulation of locomotor activity, incentive behaviors, short-term and stimulus-dependent memory systems [1,2,3]. In order to target the dopaminoceptive neurons, we have generated a transgenic line that expresses Cre recombinase in dopaminoceptive neurons under the control of the dopamine D1 receptor gene (D1Cre), by using the genomic locus of a dopamine D1 receptor gene cloned in a YAC vector
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