The dopamine transporter (DAT) protein is selectively expressed in a population of dopaminergic neurons mostly concentrated in the midbrain. Loss of DAT-expressing cells is often a marker of neurodegeneration associated with Parkinson’s disease and methamphetamine abuse. We have generated and characterized a strain of transgenic Long Evans rats expressing iCre recombinase under a DAT promoter (DAT::iCre, line 6). The tissue-specific expression of iCre can be used in combination with Cre-dependent transgenes to obtain selective expression of transgenes in DAT(+) neurons.
Status and Availability
This strain has not yet been published.
As of May 17, 2016, this strain is available as line #00758 at the RRRC.
This rat is registered at the Rat Genome Database (RGD) ID# 9588578.
Figure 1: Schematic of DAT-iCre transgenic construct. A bacterial artificial chromosome (BAC) containing the rat DAT gene (CH230-275E16) was obtained from the Children’s Hospital Oakland Research Institute (CHORI), and recombineered to replace the start codon of DAT with cassette containing iCre (improved Cre recombinase) and the polyadenylation signal from the gene for bovine growth hormone (pOTTC173). This BAC was injected into the pronuclei of fertilized Long Evans rat embryos by NIMH Transgenic Core, and ultimately resulted in three independent, phenotypically positive DAT::iCre lines. This line (LE-Tg(DAT-iCre)6Ottc) has one copy of the transgene per haploid genome as determined by droplet digital PCR.
Figure 2: RNA expression patterns of DAT (red) and iCre (green) in the midbrain. Labeling of target mRNA was done by in situ hydridization with fluorescently-labeled RNAscope probes. Wildtype rats show no iCre expression, whereas transgenic DAT-iCre6 rats show iCre expression only in cells that also express DAT. The nuclei of all cells are stained with DAPI (blue). Scale bar represents 50 microns.
Figure 3. Midbrain expression of tyrosine hydroxylase (TH) and red to green fluorescent protein reporters of Cre activity. Two weeks after injection of AAV-Nuc-flox(mCherry)-eGFP into the substantia nigra of a DAT-iCre6 rat brains were sections and immunostained for TH as a reliable marker for dopaminergic cells. Transduced cells that do not express iCre are red because they only express nuclear-localized mCherry, whereas transduced (dopaminergic) cells which express iCre are green because the viral genome has recombined to delete the mCherry coding region. The nuclei of all cells are stained with DAPI (blue). Scale bar represent 50 microns.
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