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Col1a1-driven transgenic markers of osteoblast lineage progression.
J Bone Miner Res. 2001 Jul; 16(7):1228-36.JB

Abstract

The modular organization of the type I collagen promoter allows creation of promoter-reporter constructs with preferential activity in different type I collagen-producing tissues that might be useful to mark cells at different stages of osteoblastic differentiation. Primary marrow stromal cell (MSC) and mouse calvarial osteoblast (mCOB) cultures were established from transgenic mice harboring different Col1a1 promoter fragments driving chloramphenicol acetyltransferase (CAT). In these models, Col1a1 messenger RNA (mRNA) and alkaline phosphatase (ALP) are the first markers of differentiation appearing soon after the colonies develop. Bone sialoprotein (BSP) is detected 2-3 days later, followed by osteocalcin (OC) expression and nodule mineralization. A 3.6 Col1a1 fragment (ColCAT3.6) initiated activity concomitant with ALP staining and type I collagen mRNA expression. In contrast, a 2.3 Col1a1 fragment (ColCAT2.3) became active coincident with BSP expression. The pattern of transgene expression assessed by immunostaining was distinctly different. ColCAT3.6 was expressed within and at the periphery of developing nodules whereas the ColCAT2.3 expression was restricted to the differentiated nodules. The feasibility of using green fluorescent protein (GFP) as a marker of osteoblast differentiation was evaluated in ROS17/2.8 cells. A 2.3-kilobase (kb) Col1a1 promoter driving GFP (pOB4Col2.3GLP) was stably transfected into the cell line and positive clones were selected. Subcultures lost and then regained GFP expression that was localized in small clusters of cells throughout the culture. This suggests that expression from the 2.3-kb Col1A1 fragment is determined by the state of differentiation of the ROS17/2.8 cells. Col1a1 transgenes should be useful in appreciating the heterogeneity of a primary or immortalized culture undergoing osteoblastic differentiation.

Authors+Show Affiliations

Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington 06030, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11450698

Citation

Dacic, S, et al. "Col1a1-driven Transgenic Markers of Osteoblast Lineage Progression." Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research, vol. 16, no. 7, 2001, pp. 1228-36.
Dacic S, Kalajzic I, Visnjic D, et al. Col1a1-driven transgenic markers of osteoblast lineage progression. J Bone Miner Res. 2001;16(7):1228-36.
Dacic, S., Kalajzic, I., Visnjic, D., Lichtler, A. C., & Rowe, D. W. (2001). Col1a1-driven transgenic markers of osteoblast lineage progression. Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research, 16(7), 1228-36.
Dacic S, et al. Col1a1-driven Transgenic Markers of Osteoblast Lineage Progression. J Bone Miner Res. 2001;16(7):1228-36. PubMed PMID: 11450698.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Col1a1-driven transgenic markers of osteoblast lineage progression. AU - Dacic,S, AU - Kalajzic,I, AU - Visnjic,D, AU - Lichtler,A C, AU - Rowe,D W, PY - 2001/7/14/pubmed PY - 2002/1/19/medline PY - 2001/7/14/entrez KW - Non-programmatic SP - 1228 EP - 36 JF - Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research JO - J. Bone Miner. Res. VL - 16 IS - 7 N2 - The modular organization of the type I collagen promoter allows creation of promoter-reporter constructs with preferential activity in different type I collagen-producing tissues that might be useful to mark cells at different stages of osteoblastic differentiation. Primary marrow stromal cell (MSC) and mouse calvarial osteoblast (mCOB) cultures were established from transgenic mice harboring different Col1a1 promoter fragments driving chloramphenicol acetyltransferase (CAT). In these models, Col1a1 messenger RNA (mRNA) and alkaline phosphatase (ALP) are the first markers of differentiation appearing soon after the colonies develop. Bone sialoprotein (BSP) is detected 2-3 days later, followed by osteocalcin (OC) expression and nodule mineralization. A 3.6 Col1a1 fragment (ColCAT3.6) initiated activity concomitant with ALP staining and type I collagen mRNA expression. In contrast, a 2.3 Col1a1 fragment (ColCAT2.3) became active coincident with BSP expression. The pattern of transgene expression assessed by immunostaining was distinctly different. ColCAT3.6 was expressed within and at the periphery of developing nodules whereas the ColCAT2.3 expression was restricted to the differentiated nodules. The feasibility of using green fluorescent protein (GFP) as a marker of osteoblast differentiation was evaluated in ROS17/2.8 cells. A 2.3-kilobase (kb) Col1a1 promoter driving GFP (pOB4Col2.3GLP) was stably transfected into the cell line and positive clones were selected. Subcultures lost and then regained GFP expression that was localized in small clusters of cells throughout the culture. This suggests that expression from the 2.3-kb Col1A1 fragment is determined by the state of differentiation of the ROS17/2.8 cells. Col1a1 transgenes should be useful in appreciating the heterogeneity of a primary or immortalized culture undergoing osteoblastic differentiation. SN - 0884-0431 UR - https://www.unboundmedicine.com/medline/citation/11450698/Col1a1_driven_transgenic_markers_of_osteoblast_lineage_progression_ L2 - https://doi.org/10.1359/jbmr.2001.16.7.1228 DB - PRIME DP - Unbound Medicine ER -