Abstract
The development of BAC transgenic mice expressing promoter-specific fluorescent reporter proteins has been a great asset for neuroscience by enabling detection of neuronal subsets in live tissue. For the study of basal ganglia physiology, reporters driven by type 1 and 2 dopamine receptors have been particularly useful for distinguishing the two classes of striatal projection neurons – striatonigral and striatopallidal. However, emerging evidence suggests that some of the transgenic reporter lines may have suboptimal features. The ideal transgenic reporter line should (1) express a reporter with high sensitivity and specificity for detecting the cellular subset of interest and that does not otherwise alter the biology of the cells in which it is expressed, and (2) involve a genetic manipulation that does not cause any additional genetic effects other than expression of the reporter. Here we introduce a new BAC transgenic reporter line, Drd1a-tdTomato line 6, with features that approximate these ideals, offering substantial benefits over existing lines. In this study, we investigate the integrity of dopamine-sensitive behaviors and test the sensitivity and specificity of tdTomato fluorescence for identifying striatonigral projection neurons in mice. Behaviorally, hemizygous Drd1a-tdTomato line 6 mice are similar to littermate controls; while hemizygous Drd2-EGFP mice are not. In characterizing the sensitivity and specificity of line 6 mice, we find that both are high. The results of this characterization indicate that line 6 Drd1a-tdTomato+/− mice offer a useful alternative approach to identify both striatonigral and striatopallidal neurons in a single transgenic line with a high degree of accuracy.
Highlights
The generation of transgenic lines with promoter-specific fluorescent reporter proteins has significantly advanced the field of neurobiological research
BAC transgenic mice generated by the Gene Expression Nervous System Atlas (GENSAT) project expressing EGFP fluorescent reporters under the control of the type 1 and 2 dopamine receptor regulatory sequences have been especially useful for discriminating between the two types of striatal projections neurons, striatonigral or direct pathway and striatopallidal or indirect pathway (Gong et al, 2003)
While the GENSAT Drd1a-EGFP and Drd2-EGFP transgenic lines have been useful to study the physiology of striatopallidal and striatonigral medium spiny neurons (MSNs), a recent study reports that that introduction of the BAC Drd2-EGFP transgene into the mouse genome may alter dopamine D2 receptor surface expression and cause aberrant locomotor behavior (Kramer et al, 2011)
Summary
The generation of transgenic lines with promoter-specific fluorescent reporter proteins has significantly advanced the field of neurobiological research. BAC transgenic mice generated by the Gene Expression Nervous System Atlas (GENSAT) project expressing EGFP fluorescent reporters under the control of the type 1 and 2 dopamine receptor regulatory sequences have been especially useful for discriminating between the two types of striatal projections neurons, striatonigral or direct pathway and striatopallidal or indirect pathway (Gong et al, 2003). These two types of medium spiny neurons (MSNs) have molecular, anatomical, and functional differences, but are not otherwise distinguished. These findings raise the need for alternative methodologies to discriminate between these two populations of MSNs
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