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Cloning, characterization and expression of a functional mouse ACTH receptor
F M Cammas1, S Kapas, S Barker
1Department of Chemical Endocrinology, St Bartholomew's Hospital Medical College, London, United Kingdom.
Biochemical and Biophysical Research Communications
|July 26, 1995
Summary
Researchers cloned the mouse ACTH receptor gene, demonstrating its expression and function in HeLa cells. This finding advances understanding of the ACTH receptor (ACTH-R) in mammals.
Area of Science:
- Molecular Biology
- Endocrinology
- Genetics
Background:
- The adrenocorticotropic hormone receptor (ACTH-R) plays a crucial role in regulating adrenal gland function.
- Understanding the molecular basis of ACTH-R signaling is essential for studying hormonal responses.
Purpose of the Study:
- To clone and characterize the mouse ACTH receptor gene.
- To demonstrate the functional expression of the mouse ACTH receptor in a heterologous system.
Main Methods:
- Screening a mouse genomic library using a polymerase chain reaction-generated mouse ACTH-R probe.
- Isolating and sequencing the full coding sequence of the mouse ACTH receptor gene.
- Expressing the gene in HeLa cells and measuring cyclic AMP (cAMP) production in response to ACTH and alpha-melanocyte-stimulating hormone (alpha-MSH).
Main Results:
- A 13kb clone containing the entire coding sequence of the mouse ACTH receptor gene was isolated.
- The mouse ACTH receptor gene exhibits 84% DNA homology with the human gene and encodes a peptide with 89% homology to the human ACTH-R.
- The gene is expressed as a 1.8kb transcript in the mouse adrenal gland.
- Functional expression in HeLa cells showed a dose-dependent increase in cAMP production in response to ACTH (EC50 = 7 x 10(-10)M), with alpha-MSH having no effect.
Conclusions:
- The mouse ACTH receptor gene has been successfully cloned.
- The functional expression and ACTH-mediated signaling of the mouse ACTH receptor were demonstrated in HeLa cells, providing a valuable model for further research.