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LE-Tg(cFos-eGFP)2Ottc

Background

The LE-Tg(cFos-eGFP)2Ottc rat is a reporter rat expressing enhanced green fluorescent protein (eGFP) in cells that are activated to express the immediate early gene Fos.  This rat was made by injecting a previously described DNA construct (Barth et al J Neurosci 2004; map shown below) into fertilized eggs from Long Evans rats.  Founders with incorporated transgene were screened for eGFP expression.  This line can be compared to “c-Fos-GFP” transgenic rat (described by Cifani et al. 2012, J Neurosci) as it is an independent founder from the injection mix, however LE-Tg(cFos-eGFP)2Ottc rats show more expression in caudal brain areas compared to the “c-Fos-GFP” rat described by Cifani et al. 

Status and Availability

This strain has not yet been published. 
This strain is available as line #00766 at the RRRC.
This rat is registered at the Rat Genome Database (RGD) as RGD ID# 9588570.

Transgene Information

Transgene Information/Description

Figure 1.  The schematic of the cFos-eGFP transgene showing the mouse cFos promoter as well as the first 4 exons of the cFos transcript translationally-fused to enhanced green fluorescent protein (eGFP) followed by a bovine growth hormone polyadenylation signal.  This fragment was liberated from the plasmid “c-fos-gfp” (a gift from Dr. Alison Barth, Carnegie Mellon University, Pittsburgh, PA) using HindIII and KpnI restriction enzymes, and then injected into pronuclei of fertilized Long Evans rat embryos by NIMH Transgenic Core.   Two founders were identified that carry the transgene but only one showed a eGFP-expressing phenotype.  The copy number and integration(s) have not been characterized for LE-Tg(cFos-eGFP)2Ottc.

Phenotypic Characterization


Phenotypic Characterization

Figure 2.  Activation of endogenous cFos expression and eGFP expression in prefrontal cortex.  Male LE-Tg(cFos-eGFP)2Ottc rats were injected with saline, methamphetamine (METH; 2.5 mg/kg) or haloperidol (HAL; 1 mg/kg) and immediately placed into home cage or novel environment. Two hours later, animals were perfused with 4% PFA and cryosectioned (40 μm).  Sections were immunostained for cFos overnight at 4C.  Two hours of secondary antibody.  Immunolabeled sections were mounted on slides and imaged for eGFP fluorescence and cFos immunoreactivity using NIKON upright microscope equipped with epifluorescence.  Scale bar – 100 μm. Click for larger image.

Phenotypic Characterization

Figure 3.  Activation of endogenous cFos expression and eGFP expression in dorsal striatum.  Male LE-Tg(cFos-eGFP)2Ottc rats were injected with saline, methamphetamine (METH; 2.5 mg/kg) or haloperidol (HAL; 1 mg/kg) and immediately placed into home cage or novel environment. Two hours later, animals were perfused with 4% PFA and cryosectioned (40 μm).  Sections were immunostained for cFos overnight at 4C.  Two hours of secondary antibody.  Immunolabeled sections were mounted on slides and imaged for eGFP fluorescence and cFos immunoreactivity using NIKON upright microscope equipped with epifluorescence.  Scale bar – 100 μm. Click for larger image.

Phenotypic Characterization

Figure 4.  Activation of endogenous cFos expression and eGFP expression in hippocampus.  Male LE-Tg(cFos-eGFP)2Ottc rats were injected with saline, methamphetamine (METH; 2.5 mg/kg) or haloperidol (HAL; 1 mg/kg) and immediately placed into home cage or novel environment. Two hours later, animals were perfused with 4% PFA and cryosectioned (40 μm).  Sections were immunostained for cFos overnight at 4C.  Two hours of secondary antibody.  Immunolabeled sections were mounted on slides and imaged for eGFP fluorescence and cFos immunoreactivity using NIKON upright microscope equipped with epifluorescence.  Scale bar – 100 μm. Click for larger image.

Experiments conducted at the NIDA IRP by Bruce Hope, YaJun Zhang and Brandon Harvey.

Additional phenotypic characterization is recommended for brain region and experimental paradigm of interest.

Breeding Strategy:

Breeding Information, click HERE

Genotyping Assays  

Protocol for genotyping transgenic rats, 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:

LE-Tg(cFos-eGFP)1Ottc  (aka “c-Fos-GFP” rat)
Published as the “c-Fos-GFP” rat which is also a randomly integrated transgene on a Long Evans background that is described by Cifani et al 2012 J Neurosci.  If you are interested in the “c-Fos-GFP” rat, please contact Bruce Hope (bhope@intra.nida.nih.gov).

Acknowledgements:

YaJun Zhang, Julie Necarsulmer, Christopher T. Richie, Brandon Harvey, Janette Lebron