Characterization and identification of corticotropin-releasing hormone receptors.
Project/Area Number |
03671146
|
Research Category |
Grant-in-Aid for General Scientific Research (C)
|
Allocation Type | Single-year Grants |
Research Field |
内分泌・代謝学
|
Research Institution | KYOTO UNIVERSITY |
Principal Investigator |
FUKATA Junichi Kyoto University Faculty of Medicine, Instructor, 医学部, 助手 (20189913)
|
Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Hiromasa Kyoto University Faculty of Medicine, Graduate Student, 医学部, 大学院生
|
Project Period (FY) |
1991 – 1992
|
Project Status |
Completed (Fiscal Year 1992)
|
Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1992: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1991: ¥1,600,000 (Direct Cost: ¥1,600,000)
|
Keywords | Corticotropin-releasing hormone / Receptor / 副腎皮質刺激ホルモン放出因子 |
Research Abstract |
As a first step to clarify corticotropin-releasing hormone receptor (CRHR), we first studied putative receptors for cytokines on a mouse pituitary tumor cell line AtT-20, which is known to respond to CRH to synthesize and release corticotropin(ACTH)-related peptides. Binding assays using ^<125>I-labeled interleukin(IL)-1 showed the cells had little but typical receptors to IL-1. Similarly, ^<125>I-labeled tumor necrosis factor (TNF)-alfa also showed the existence of its receptor molecules on AtT-20 cells. Using labeled cDNA for TNF-alfa receptor molecule, we showed the expression of TNF-alfa-mRNA in the cells. Next, we study an aliquot of 1-biotin-labeled CRH binding on the AtT- 20 cells similarly. Although 1-biotin-labeled CRH, which we synthesized for the binding study, showed comparable capacity to release ACTH from the AtT-20 cells, we could not obtain positive binding of the labeled CRH on the AtT-20 cells using flow cytometry analysis. Since we had generated specific and high affinity receptor against CRH, we also tried to study whether the antibody was suitable to detect cells with CRH receptors on their surface with anti-CRH-coated ELISA plates but also failed positive panning study. Further study should be necessarily continued to finally elucidate the nature of CRHR molecules.
|
Report
(3 results)
Research Products
(10 results)