Analysis of thyroid hormone responsive gene expression in osteoblastic cells.
Thyroid hormones regulate gene expression to influence the development and metabolism of many tissues including bone. The identification of genes that are regulated by thyroid hormones during skeletal development requires sensitive and quantitative techniques that are not limited by small amounts of available tissue and RNA. We have compared the efficiencies of differential display and poly A PCR subtraction hybridisation methods for the detection of thyroid hormone responsive genes expressed in osteoblastic cells. The utility of each technique was evaluated with respect to its sensitivity, specificity, cost and ability to identify novel genes. Subtraction hybridisation was rapid and more efficient in all categories. Poly A PCR facilitates quantitative and representative global amplification of cDNAs from low concentrations of RNA extracted from small tissue samples. The method, in combination with microarray analyses, may prove useful as an additional, complementary strategy to subtraction hybridisation for the analysis of differential gene expression in tissues where sample size is limiting.
Harvey
C B
CB
Division of Medicine and MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK.
Stevens
D A
DA
Williams
A J
AJ
Jackson
D J
DJ
O'Shea
P
P
Williams
G R
GR
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000032:analysis; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D015972:Gene Expression Regulation, Neoplastic; D010006:Osteoblasts / Q000378:metabolism*; D011061:Poly A; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis; D051381:Rats; D012680:Sensitivity and Specificity; D014284:Triiodothyronine / Q000502:physiology*
D018076:DNA, Complementary; D014284:Triiodothyronine; D011061:Poly A; D012313:RNA