Related Experiment Videos
Oestrogen receptors in macrophages
S Gulshan1, A B McCruden, W H Stimson
1Department of Bioscience and Biotechnology, University of Strathclyde, Glasgow, Scotland, UK.
Scandinavian Journal of Immunology
|June 1, 1990
Summary
Researchers investigated oestrogen receptors in J111 cells and rat macrophages. They found oestrogen-binding species but no 5 alpha-dihydrotestosterone (5 alpha-DHT) receptors, suggesting a unique macrophage oestrogen receptor type.
Area of Science:
- Endocrinology
- Cell Biology
- Immunology
Background:
- Steroid hormones like oestrogen play crucial roles in cellular function.
- Macrophages are key immune cells involved in inflammatory and immune responses.
- Understanding hormone receptor expression in immune cells is vital for comprehending their regulation.
Purpose of the Study:
- To investigate the presence and characteristics of oestrogen and 5 alpha-dihydrotestosterone (5 alpha-DHT) receptors in the J111 human monocytic leukaemia cell line and rat peritoneal macrophages.
- To determine the binding affinities and properties of any identified oestrogen-binding species.
Main Methods:
- Whole-cell binding assays were employed to detect receptor presence and binding affinity.
- Scatchard analysis was used to characterize binding kinetics and identify receptor populations.
- Sucrose-gradient ultracentrifugation was performed to determine the sedimentation coefficient of oestrogen-binding species.
- Competitive binding assays with diethylstilboestrol (DES) and a monoclonal antibody against classical oestrogen receptors were conducted.
Main Results:
- High-affinity oestrogen-binding species were detected in both J111 cells and rat peritoneal macrophages.
- No specific binding for 5 alpha-dihydrotestosterone (5 alpha-DHT) was observed in either cell type.
- Scatchard analysis revealed two distinct oestrogen-binding species in both cell lines, with calculated dissociation constants (Kd) in the nanomolar range.
- Sucrose-gradient ultracentrifugation identified a single 7.1S oestrogen-binding species.
- The receptors exhibited high affinity for diethylstilboestrol (DES) but did not react with an antibody specific for classical oestrogen receptors.
Conclusions:
- J111 cells and rat peritoneal macrophages possess oestrogen receptors but lack detectable 5 alpha-DHT receptors.
- The presence of two oestrogen-binding species suggests complex receptor regulation or distinct receptor subtypes.
- The 7.1S oestrogen-binding species and its differential reactivity suggest a non-classical oestrogen receptor type in macrophages.