Characterization of Pdgfrb‐Cre transgenic mice reveals reduction of ROSA26 reporter activity in remodeling arteries

Abstract

With the intention to modulate gene expression in vascular mural cells of remodeling vessels, we generated and characterized transgenic mouse lines with Cre recombinase under the control of the platelet-derived growth factor receptor-β promoter, referred to as Tg(Pdgfrb-Cre)35Vli. Transgenic mice were crossed with the Gt(ROSA)26Sortm1Sor strain and examined for Cre activation by β-galactosidase activity, which was compared with endogenous Pdgfrb expression. In addition, Pdgfrb-Cre mice were used to drive expression of a conditional myc-tagged Cthrc1 transgene. There was good overlap of β-galactosidase activity with endogenous Pdgfrb immunoreactivity. However, dedifferentiation of vascular mural cells induced by carotid artery ligation revealed a dramatic discrepancy between ROSA26 reporter activity and Pdgfrb promoter driven Cre dependent myc-tagged Cthrc1 transgene expression. Our studies demonstrate the capability of the Pdgfrb-Cre mouse to drive conditional transgene expression as a result of prior Cre mediated recombination in tissues known to express endogenous Pdgfrb. In addition, the study shows that ROSA26 promoter driven reporter mice are not suitable for lineage marking of smooth muscle in remodeling blood vessels.

Dynamic Remodeling of Pericytes In Vivo Maintains Capillary Coverage in the Adult Mouse Brain

“…A single intraperitoneal injection of tamoxifen (100 mg/kg) to either mouse line led to sparse labeling of mural cells throughout the brain. The third transgenic mouse line achieved constitutive labeling of all mural cells through the cell-specific expression of YFP (Ai3)(Madisen et al, 2010) under the PDGFRβ promoter (Cuttler et al, 2011). We will refer to these as PDGFRβ-YFP mice.…”

“…019079). In a subset of pericyte ablation and BBB leakage experiments, we crossed constitutive PDGFRβ-Cre mice (Cuttler et al, 2011) with a YFP reporter line (Ai3)(Madisen et al, 2010) to express YFP in all mural cells. Mice were maintained in standard cages on a 12-hour light-dark cycle, and housed 5 or less per cage.…”

Novel Adult Stem Cells for Peripheral Nerve Regeneration

“…3f). The lack of EGFP expression in MVSCs was unlikely due to the suppression of Rosa26 promoter activity in cell culture 33 . When SM-MHC-Cre/LoxP-EGFP mice were generated using β-actin (instead of ROSA26) promoter to drive the EGFP expression, tissue explant culture showed that cells derived from these mice did not express EGFP, while vascular cells in the tissue explant expressed EGFP (Supplementary Fig.…”

Mesangial cell αvβ8-integrin regulates glomerular capillary integrity and repair

“…PDGFRB-Cre mice [PDGFRB-Cre(RA) mice] were a gift from Dr. Ralf Adams (Cancer Research UK London Research Institute) (10). A second strain of PDGFRB-Cre mice, which contain a longer promoter with the transgene [PDGFRB-Cre(VL) mice], was a gift from Dr. Volkhard Lindner (Maine Medical Center Research Institute) (8). Ren1 d -Cre mice were a gift from Dr. Ariel Gomez (University of Virginia) (40).…”