A critical developmental role for tgfbr2 in myogenic cell lineages is revealed in mice expressing SM22-Cre, not SMMHC-Cre

Abstract

Smooth muscle cell (SMC)-specific deletion of transforming growth factor beta (TGF-β) signaling would help elucidate the mechanisms through which TGF-β signaling contributes to vascular development and disease. We attempted to generate mice with SMC-specific deletion of TGF-β signaling by mating mice with a conditional (“floxed”) allele for the type II TGF-β receptor ( tgfbr2 flox) to mice with SMC-targeted expression of Cre recombinase. We bred male mice transgenic for smooth muscle myosin heavy chain (SMMHC)-Cre with females carrying tgfbr2 flox. Surprisingly, SMMHC-Cre mice recombined tgfbr2 flox at low levels in SMC and at high levels in the testis. Recombination of tgfbr2 flox in testis correlated with high-level expression of SMMHC-Cre in testis and germline transmission of tgfbr2 null. In contrast, mice expressing Cre from a SM22α promoter (SM22-Cre) efficiently recombined tgfbr2 flox in vascular and visceral SMC and the heart, but not in testis. Use of the R26R reporter allele confirmed that Cre-mediated recombination in vascular SMC was inefficient for SMMHC-Cre mice and highly efficient for SM22-Cre mice. Breedings that introduced the SM22-Cre allele into tgfbr2 flox/flox zygotes in order to generate adult mice that are hemizygous for SM22-Cre and homozygous for tgfbr2 flox- and would have conversion of tgfbr2 flox/flox to tgfbr2 null/null in SMC-produced no live SM22-Cre : tgfbr2 flox/flox pups ( P < 0.001). We conclude: (1) “SMC-targeted” Cre lines vary significantly in specificity and efficiency of Cre expression; (2) TGF-β signaling in the subset of cells that express SM22α is required for normal development; (3) generation of adult mice with absent TGF-β signaling in SMC remains a challenge.