VCAM-1 ablation in nonhematopoietic cells in MxCre+ VCAM-1f/f mice is variable and dictates their phenotype.
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.
Ulyanova
Tatiana
T
Department of Medicine, Division of Hematology, University of Washington, Seattle, WA 98195, USA.
Priestley
Gregory V
GV
Nakamoto
Betty
B
Jiang
Yi
Y
Papayannopoulou
Thalia
T
D000818:Animals; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000378:metabolism*; D002469:Cell Separation; D017931:DNA Primers; D007150:Immunohistochemistry; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D010641:Phenotype; D016133:Polymerase Chain Reaction; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D019010:Vascular Cell Adhesion Molecule-1; C045073:Cre recombinase; D019426:Integrases