Adrenocorticotropin receptors: Functional expression from rat adrenal mRNA in Xenopus laevis oocytes
- National Inst. of Health, Bethesda, MD (United States)
The adrenocorticotropin (ACTH) receptor, which binds corticotropin and stimulates adenylate cyclase and steroidogenesis in adrenocortical cells, was expressed in Xenopus laevis oocytes microinjected with rat adrenal poly(A){sup +} RNA. Expression of the ACTH receptor in individual stage 5 and 6 oocytes was monitored by radioimmunoassay of ligand-stimulated cAMP production. Injection of 5-40 ng of adrenal mRNA caused dose-dependent increases in ACTH-responsive cAMP production. Size fractionation of rat adrenal poly(A){sup +}RNA by sucrose density-gradient centrifugation revealed that mRNA encoding the ACTH receptor was present in the 1.1-to 2.0-kilobase fraction. These data indicate that ACTH receptors can be expressed from adrenal mRNA in Xenopus oocytes and are fully functional in terms of ligand specificity and signal generation. The extracellular cAMP response to ACTH is a sensitive and convenient index of receptor expression. This system should permit more complete characterization and expression cloning of the ACTH receptor.
- OSTI ID:
- 5701147
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 88:19; ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
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ADRENAL HORMONES
RADIOIMMUNOASSAY
CYCLASES
ENZYME ACTIVITY
RECEPTORS
BIOCHEMICAL REACTION KINETICS
AMP
AMPHIBIANS
CENTRIFUGATION
CORTICOSTEROIDS
DOSE-RESPONSE RELATIONSHIPS
MESSENGER-RNA
OOCYTES
ANIMALS
AQUATIC ORGANISMS
BIOASSAY
DIAGNOSTIC TECHNIQUES
ENZYMES
GERM CELLS
HORMONES
HYDROXY COMPOUNDS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
KETONES
KINETICS
LYASES
MEMBRANE PROTEINS
NUCLEIC ACIDS
NUCLEOTIDES
ORGANIC COMPOUNDS
PREGNANES
PROTEINS
RADIOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
REACTION KINETICS
RNA
SEPARATION PROCESSES
STEROID HORMONES
STEROIDS
TRACER TECHNIQUES
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550600* - Medicine