VCAM-1 ablation in nonhematopoietic cells in MxCre+ VCAM-1f/f mice is variable and dictates their phenotype

Exp Hematol. 2007 Apr;35(4):565-71. doi: 10.1016/j.exphem.2007.01.031.

Abstract

Objective: The goal of the present study was to assess the extent of vascular cell adhesion molecule-1 (VCAM-1) gene deletion in hematopoietic vs nonhematopoietic cells in the bone marrow (BM) of MxCre(+)VCAM-1(f/f) mice and its impact on the phenotypic features of these mice.

Methods: VCAM-1 ablation was evaluated at the genomic level by polymerase chain reaction (PCR), at the mRNA level by real-time PCR, and at the protein level by fluorescein-activated cell sorting and immunohistochemistry. The homing or mobilization of colony-forming unit cultures was assessed by standard assays.

Results: A previously accepted interferon-induction scheme yielded efficient VCAM-1 ablation in hematopoietic cells but variable ablation in BM fibroblasts and endothelial cells. The level of ablation in the latter populations correlated with alterations in the hematopoietic phenotype.

Conclusions: Poly(I:C)-induced MxCre-mediated gene ablation is highly efficient in hematopoietic cells but variable and partial in nonhematopoietic cells in BM. Ablation of VCAM-1 in hematopoietic cells does not contribute to their mobilization, nor does it impair their homing. The latter is dependent on VCAM-1 ablation in nonhematopoietic cells of BM.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Base Sequence
  • Bone Marrow Cells / metabolism*
  • Cell Separation
  • DNA Primers
  • Immunohistochemistry
  • Integrases / genetics
  • Integrases / metabolism*
  • Mice
  • Phenotype
  • Polymerase Chain Reaction
  • Vascular Cell Adhesion Molecule-1 / genetics
  • Vascular Cell Adhesion Molecule-1 / metabolism*

Substances

  • DNA Primers
  • Vascular Cell Adhesion Molecule-1
  • Cre recombinase
  • Integrases