Development of inducible cell model systems to study MT 1 and MT2 melatonin receptors.

摘要

Melatonin, N-acetyl-5-methoxytryptamine, is a naturally occurring hormone that is secreted from the pineal gland into the bloodstream during the hours of darkness. Researchers have studied the function of melatonin in humans in an effort to elucidate its many functions and its role in the regulation of circadian rhythms. Previous research has utilized transformed Chinese Hamster Ovary cells (CHO) that constitutively express the MT₁ and MT ₂ receptor genes. In this study, we created four cell lines with inducible MT₁ and MT₂ receptor expression in both non-neuronal (ind-MT₁-CHO-CL6, ind-MT₂-CHO-B) and neuronal (ind-MT ₁-BE(2)C-CL1, ind-MT₂-BE(2)C-CL2) cell backgrounds. These cell lines can be induced to express receptor levels that are similar to what is seen in actual physiological human conditions. To create inducible cell lines, the Ecdysone Inducible Mammalian Expression System was used. To create a reporter molecule for monitoring protein expression and localization, a MT₁-GFP construct was created. (Abstract shortened by UMI.)