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Faculty
David W. Rowe
Professor of Genetics and Developmental Biology
rowe@neuron.uchc.edu
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. |
