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.
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.
Authors: Hanna K A Mikkola; Jenny Klintman; Haidi Yang; Hanno Hock; Thorsten M Schlaeger; Yuko Fujiwara; Stuart H Orkin Journal: Nature Date: 2003-01-19 Impact factor: 49.962
Authors: Yi Gu; Marie-Dominique Filippi; Jose A Cancelas; Jamie E Siefring; Emily P Williams; Aparna C Jasti; Chad E Harris; Andrew W Lee; Rethinasamy Prabhakar; Simon J Atkinson; David J Kwiatkowski; David A Williams Journal: Science Date: 2003-10-17 Impact factor: 47.728
Authors: Liping Liu; MeiZhang Li; Lisa C Spangler; Charles Spear; Mike Veenstra; Lindsey Darnall; Cathleen Chang; Anne C Cotleur; Richard M Ransohoff Journal: Genesis Date: 2013-06-24 Impact factor: 2.487