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DEVELOPMENT OF A GENE-EXPRESSION ARRAY FOCUSING ON THE HYPOTHALMUS-PITUARY-THYROID AXIS IN XENOPUS LAEVIS
Citation:
Korte, J J., J E. Tietge, AND S J. Degitz. DEVELOPMENT OF A GENE-EXPRESSION ARRAY FOCUSING ON THE HYPOTHALMUS-PITUARY-THYROID AXIS IN XENOPUS LAEVIS. Presented at Third Indo-US Workshop on Mathematical Chemistry, Duluth, MN, August 2-7, 2003.
Description:
As recommended by the Endocrine Disrupter Screening and Testing Program Advisory Committee (EDSTAC), the US EPA has been developing a screening test capable of detecting effects of Endocrine Disrupting Chemicals (EDCS) on the hypothalamus-pituatary-thyroid (HPT) axis in Xenopus laevis. As part of this, we developed an oligonucleotide array to assess changes in expression of important thyroid-axis genes, allowing comparison of the results with morphometric and biochemical information. The method consists of hybridizing amplified, biotinylated RNA from test organismsa with membrane-bound synthetic DNA oligomers of 65 to 70 bases. The biotib label is detected by chemiluminescence and quantified. - - - The array has been utilized on brain samples from stage 54 to 62 tadpoles to determine the pattern of gene expression in control animals during development. Future studies with known modulators of thyroid function will compare expression patterns to normal tadpoles to look for indications of altered gene expression. Ultimately, we hope to utilize the method to indicate if the thyroid axis has been impacted by potential EDCs.