Areas of Interest
The primary focus of the laboratory is to recognize and
ultimately isolate subpopulations of cells at define levels of
differentiation within the osteoprogenitor lineage. This goal
has been made possible by the construction of a series of mice
transgenic for promoter-GFP reporter genes that activate at
defined levels of osteoblast differentiation. Fluorescent
microscopic methods have been developed to observed lineage
progression in real time in culture, to map where a mutation
acts to impede forward progress of cell development and to
correlate its in vivo impact within intact mouse bone.
Particular attention is directed in multiplexing various colors
of GFP and fluorescent cellular stains to maximize the
information available within a histological section of bone.
Examples of this new approach to cellular analysis can be seen
at:
http://skeletalbiology.uchc.edu/, click image center and see
some of the images that are in current publications or at
http://skeletalbiology.uchc.edu/30_ResearchProgram/304_gap/index.htm
which details more of the methodologies.
Current projects that utilize this experimental approach
include: a) microarray analysis of genes expressed in FAC
isolated subpopulations within the osteoblast lineage; b)
examination of a murine model of osteogenesis imperfecta (OIM)
from the perspective of osteoblast lineage regulation; c)
evaluation of methods to correct a dominant negative mutation
such as OIM with various antiRNA strategies; d) effectiveness of
various osteoprogenitor transplantation protocols for osteoblast
engraftment. Graduate students will be exposed to techniques of
DNA cloning and BAC recombineering, traditional and microarray
analysis of RNA, retroviral vectors, tissue culture and
histological assessment of the osteoblast lineage and management
and analysis of murine models of human disease.
Selected Publications
Bilic-Curcic I, Kronenberg M, Jiang X, Bellizzi J, Mina M,
Marijanovic I, Gardiner EM, Rowe DW. Visualizing levels of
osteoblast differentiation by a two-color promoter-GFP strategy:
Type I collagen-GFPcyan and osteocalcin-GFPtpz. Genesis.
43(2):87-98, 2005.
Jiang X, Kalajzic Z, Maye P, Braut A, Bellizzi J, Mina M,
Rowe DW.
Histological analysis of GFP expression in murine bone. J
Histochem Cytochem. 53(5):593-602, 2005.
Visnjic, D., Kalajzic, Z., Rowe, D., Katavic, V., Lorenzo,
J., Aguila, H. L. Hematopoiesis is severely altered in mice with
an induced osteoblast deficiency. Blood 103: 3258-3264, 2004.
Kalajzic, I., Braut, A., Guo D., Xi, J., Kronenberg, M.S.,
Mina, M., Harris MA, Harris, S.E. and Rowe, D.W. Dentin Matrix
Protein 1 Expression During Osteoblastic Differentiation and a
12kb cis-regulatory region with modules specific to Osteocytes.
Bone 35:74-82, 2004.
Liu, P., Mark Kronenberg, M., Zhang, X and Rowe, D. Modified
U1snRNAs suppresses expression of a targeted endogenous RNA by
inhibiting polyadenylation of the transcript. Nuc Acid Res.
32:1512-1517, 2004.
Garg, S., Hansen, M.F., Rowe, D.W. and Achenie, L.E. An
Adaptive Strategy for Single and Multi Cluster Gene Assignment.
Biotechol. Prog. 19:1142-1148, 2003.
Kalajzic, I., Kalajzic, Z., Lichtler, A. and Rowe, D. Noggin
and FGF2 inhibit progression of the osteoprogenitor at different
stages of development. J. Cell Biochemistry, 88: 1168-1176,
2003.
Fortes, P, Cuevas, Y., Guan, F., Liu, P., Pentlicky, S.,
Jung, S., Martínez-Chantar, M., Prieto, J., Rowe, D., Gunderson,
S. Inhibiting expression of specific genes in mammalian cells
with 5' end-mutated U1 snRNAs targeted to terminal exons of
pre-mRNA. Proc. Natl.Acad.Sci. 100:8264-8269, 2003.
Plotkin, H., Primorac, D. and Rowe, D. Osteogenesis
Imperfecta. In Pediatric Bone, Biology and Diseases. Ed. by F.H.
Glorieux, J.M. Pettifor and H. Juppner. Academic Press (2003)
Chapter 18, pp 443-472.
Liu, P., Gucwa, A., Stover, M.L., Buck, E., Lichtler A. and
Rowe, D. Analysis of Inhibitory Action of Modified U1 snRNAs on
Target Gene Expression: Discrimination of two RNA targets
differing by a 1 bp mismatch. Nuc Acid Res. 30: 2329-2339, 2002.
Kalajzic, I., Kalajzic, Z, Clark, S, Lichtler, A. and Rowe,
D. Use of Col1a1GFP transgenes to identify subpopulations of
cells at different stages of the osteoblast lineage. J.Bone Min.
Res. 17: 15-25, 2002.
Braut, A., Kalajzic, I., Kalajzic, Z., Rowe, D.W., Kollar,
E.J. and Mina, M. Col1a1-GFP transgene expression in developing
incisiors. Conn. Tissue . Res. 43: 216-219, 2002.
Rowe, D. and Lichtler, A. A Strategy for Identifying
Osteoporosis Risk Genes. Endocrine, 17: 67-75, 2002.
Kalajic, I., Terzic, J., Rumboldt, Z., Mack, K., Napta, A.,
Gronowicz, G., Ledgard, F., Clark, S. and Rowe, D. Osteoblastic
response to the defective matrix in the osteogenesis imperfecta
murine (oim) mouse. Endocrinology, 143:1594-1601, 2002.
Visnjic, D., Kalajzic, I., Gronowicz, G., Clark, S. Lichtler,
A. and Rowe, D. The Col2.3∆TK mouse: a model for conditional
ablation of the osteoblast lineage. J. Bone Min. Res. 16:
2222-2231, 2001.
Liu, P., Kalajzic, I., Stover, M-L, Rowe, D. and Lichtler, A.
Human bone marrow stromal cells are efficiently transduced by
VSV pseudotyped retrovectors without affecting subsequent
osteoblastic differentiation. Bone 29:331-335, 2001.
Stover, M-L., Wang, C-K, McKinstry, M., Kalajzic, I.,
Gronowicz, G., Clark, S., Rowe, D. and Lichtler, A. Bone
directed expression of Col1A1 promoter-driven self-inactiviating
retroviral vector in bone marrow cells and transgenic mice. Mol
Ther. 3:543-549, 2001.
Beckley, A., Liu, P., Stover, M.L. Gunderson, S.I., Lichtler,
A. and Rowe, D.W. Reduction of target gene expression by a
modified U1 snRNA. Mol Cell Biol. 2815-2825, 2001.
Dacic, S., Kalajzic, I, Visnjic, D., Lichtler, A. and Rowe,
D. Col1A1 driven transgenic markers of osteoblast lineage
progression. J.Bone Min. Res. 16:1228-1236, 2001.
Kalajzic, I., Kalajzic, Z, Liu, P., Stover, M-L., Rowe, D.
and Lichtler, A. Use of VSV-G pseudotyped retroviral vectors to
target murine osteoprogenitor cells Virology 284:37-45, 2001. |
