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LE-Tg(DAT-iCre)6Ottc

Background

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.



Transgene Information

Legend: 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.

 

Phenotypic Characterization

Legend: Images showing the RNA expression patterns of DAT (red) and iCre (green) in the midbrain after 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.


Legend:  Images showing expression patterns of tyrosine hydroxylase (TH) and fluorescent protein reporters two weeks after injection of AAV-Nuc-flox(mCherry)-eGFP into the substantia nigra of a DAT-iCre6 rat.  Staining with anti-TH antibodies is 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.

Breeding Strategy:

Breeding Information, click HERE

Genotyping Assays  

Assay for the presence of DAT-iCre insert, click HERE

Assay for the presence of iCre insert, click HERE

References that cite this rat:

No references are available at this time.

Blog/Comments/Reviews

No comments or reviews are available at this time.

Other related rats:

Registered Symbol

RGD ID

Proposed Name

RRRC#*

LE-Tg(DAT-iCre)1Ottc 9588572 LE-Tg(DAT::iCre)1Ottc 730
LE-Tg(DAT-iCre)5Ottc 9588576 LE-Tg(DAT::iCre)5Ottc N/A

 

These rat lines also resulted from the generation of LE-Tg(DAT-iCre)6Ottc described above. Line 1 has 2 copies of transgene and Line 5 has 8 copies of transgene. These lines did show expression of Cre in midbrain dopamine neurons and promoted recombination as tested by Cre-dependent reporters delivered by AAV vectors. Line 6 was chosen as primary line because it had a single transgene copy per genome and showed appropriate phenotype. Should alternative lines be required in the future, these lines have been cryopreserved. *Rat Research and Resource Center (RRRC)