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Text
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URL Address
<a href="http://doi.org/10.1080/03008200390152052" target="_blank" rel="noreferrer noopener">http://doi.org/10.1080/03008200390152052</a>
Pages
28–32
Volume
44 Suppl 1
Dublin Core
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Title
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Analysis of osteopontin in mouse growth plate cartilage by application of laser capture microdissection and RT-PCR.
Publisher
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Connective tissue research
Date
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2003
1905-06
Subject
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*Reverse Transcriptase Polymerase Chain Reaction; Animals; Articular/chemistry/cytology; Cartilage; Chondrocytes/chemistry; Gene Expression; Growth Plate/*chemistry/cytology; Inbred C57BL; Laser Therapy; Messenger/metabolism; Mice; Microdissection/*methods; Newborn; Non-programmatic; Osteopontin; RNA; Sialoglycoproteins/*analysis/genetics; Tibia
Creator
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Landis William J; Jacquet Robin; Hillyer Jennifer; Zhang Jean
Description
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Gene expression of osteopontin (OPN) has been investigated in mice by application of laser capture microdissection (LCM) and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. LCM permits individual cells to be isolated ("captured") from tissue sections for molecular analyses. In this study, chondrocytes were captured from growth plate zones in frozen sections of tibiae from 1-11-day-old postnatal mice. RNA was extracted from cells, DNAse-treated, and reverse-transcribed. cDNA was amplified by PCR and OPN mRNA was revealed on agarose gels. Whole cartilage and brain (a positive control) from the same animals also were examined. Reactions containing no RT were negative controls, and 18S rRNA standardized expressed message from captured cells. RT-PCR analysis of laser-captured whole cartilage showed a general qualitative loss of OPN mRNA as animal age increased. Youngest mice gave equivalent OPN expression over all laser-microdissected cartilage zones. For 7-11 day-old mice, OPN expression was qualitatively greatest in resting and lowest in hypertrophic regions of cartilage. Expression of OPN correlated with mineral in the tissue suggests that OPN functionally may inhibit normal vertebrate growth plate mineralization, and its loss with increasing tissue maturation appears permissive to mineral development.
Identifier
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<a href="http://doi.org/10.1080/03008200390152052" target="_blank" rel="noreferrer noopener">10.1080/03008200390152052</a>
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Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
*Reverse Transcriptase Polymerase Chain Reaction
2003
Animals
Articular/chemistry/cytology
Cartilage
Chondrocytes/chemistry
Connective tissue research
Gene Expression
Growth Plate/*chemistry/cytology
Hillyer Jennifer
Inbred C57BL
Jacquet Robin
Landis William J
Laser Therapy
Messenger/metabolism
Mice
Microdissection/*methods
Newborn
Non-programmatic
Osteopontin
RNA
Sialoglycoproteins/*analysis/genetics
Tibia
Zhang Jean