Analysis of thyroid stimulating hormone‐receptor mutations by temperature‐gradient gel electrophoresis

Abstract

Somatic mutations in the genes for G-protein-coupled receptors which regulate intracellular levels of cyclic AMP have been found in several regions coding for the receptors of melanocyte-stimulating hormone, adrenaline, luteinizing hormone, rhodopsin and thyrotropin. The mutations found in the thyroid-stimulating hormone (TSH) receptor are restricted to cells of hyperfunctioning thyroid adenomas and other thyroid tumors. They are thought to lead to a constitutive activation of the receptor independent of TSH. We have developed a temperature-gradient gel electrophoresis (TGGE) assay starting with the amplification of a part of exon 10 of the TSH-receptor gene to screen tissue of thyroid tumours. TGGE allows the detection of point mutations even if there are only a few cells with somatic mutations in tissue sections.

Double-Gradient DGGE for Optimized Detection of DNA Point Mutations

“…Notwithstanding the more cumbersome experimental setup (often requiring the use of two thermostats for generating the temperature gradient and the use of special precautions for avoiding unequal solvent evaporation from the gel slab), TGGE is also gaining in popularity, as witnessed by bursts of reports on this methodology (17,18,24,25). A similar method in a capillary format is being developed (11).…”