Easy detection of hormone secretion from LbetaT2 cells by using gaussia luciferase
2017
Satou, K. (Meiji University, Kanagawa (Japan). School of Agriculture, Department of Life Sciences, Laboratory of Cell Signaling Regulation) | Mochimaru, Y. | Nakakura, T. | Kusada, T. | Negishi, J. | Musha, S. | Yoshimura, N. | Kato, Y. | Tomura, H.
Reproduction is regulated by gonadotropins secreted from gonadotrophs. The production and secretion of gonadotropins are mainly regulated by gonadotropin-releasing hormone (GnRH). Agonists or antagonists that influence GnRH action on gonadotrophs are important to regulate reproduction; however, these factors have not been fully characterized due to the lack of simple and easy-to-use techniques to detect gonadotropin secretion from gonadotropin-producing cells. In the present study, we found that Gaussia luciferase (Gluc), which was expressed in LbetaT2 cells, can be secreted like a luteinizing-hormone (LH) upon stimulation with GnRH. The Gluc secreted into the medium was easily monitored as luminescence signals. The detection range of the GnRH-induced Gluc activity was comparable to that of the enzyme-linked immunosorbent assay for LH. In addition, when the Gluc was expressed in AtT20 cells, which produce adrenocorticotropic hormone (ACTH), the Gluc activity in the medium increased in parallel with the ACTH secretion upon stimulation with corticotropin-releasing hormone. Thus, the Gluc assay in the present study can be easily used for high-throughput screening of factors that influence LH or ACTH secretion from LbetaT2 or AtT20 cells, respectively.
Показать больше [+] Меньше [-]