Gene expression in skeletal tissues: application of laser capture microdissection.
Tissue differentiation is based on the expression of transcription factors, receptors for cytokines, and nuclear receptors that regulate a specific phenotype. The purpose of this study was to select cells from various skeletal tissues in order to analyse differential gene expression of cells in the native environment in vivo. It is a difficult task to obtain cells from skeletal tissues, such as cartilage, periost, bone and muscle, that are structured together and do not exist as individual organs. We used laser capture microdissection which permits the selection and isolation of individual cells from tissue sections. The RNA isolated from these tissues was used for reverse transcriptase-polymerase chain reactions for molecular analysis. We analysed the expression of transcription factors (cFOS, cbfa1, MyoD), receptors for cytokines, nuclear receptors, alkaline phosphatase and the structural proteins osteocalcin and collagen II. The results obtained demonstrate differential patterns of gene expression according to the tissue arrangement in their native in vivo environment, with reliable interpretation of the functions of the analysed genes in the context of intact skeletal tissue physiology.
Benoyahu
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D
Department of Cell and Developmental Biology, Sackler School of Medicine, Tel-Aviv University, Israel. dafnab@post.tau.ac.il
Akavia
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UD
Socher
R
R
Shur
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I
D000818:Animals; D001842:Bone and Bones / Q000378:metabolism*; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D015870:Gene Expression; D051379:Mice; D008810:Mice, Inbred C57BL; D042282:Microdissection; D017570:MyoD Protein / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D018121:Receptors, Cytokine / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics; D011965:Receptors, Glucocorticoid / Q000235:genetics
D050659:Core Binding Factor Alpha 1 Subunit; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D016760:Proto-Oncogene Proteins c-fos; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D011960:Receptors, Estrogen; D011965:Receptors, Glucocorticoid