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Published on: April 3, 2014
Estradiol effects on the dopamine transporter - protein levels, subcellular location, and function
Cheryl S Watson1, Rebecca A Alyea, Bridget E Hawkins
1Department of Biochemistry & Molecular Biology, Univ, of Texas Medical Branch, Galveston TX 77555-0645, USA. cswatson@utmb.edu
Journal of Molecular Signaling
|January 17, 2007
Summary
Estrogen rapidly inhibits dopamine transporter (DAT) activity in PC12 cells, suggesting a membrane-initiated mechanism. This rapid estrogenic action may involve sequestering DAT within intracellular compartments via membrane estrogen receptors.
Area of Science:
- Neuroscience
- Endocrinology
- Molecular Biology
Background:
- Estrogen's influence on dopamine transport is crucial for understanding neurological disorders in women.
- Hormonal fluctuations during key life stages may impact neurological health.
Purpose of the Study:
- To investigate the rapid, non-genomic effects of 17beta-estradiol (E2) on dopamine transporter (DAT) activity.
- To explore the role of membrane estrogen receptors (ERs) in mediating these rapid effects.
Main Methods:
- Measured 3H-DA uptake in PC12 cells treated with varying concentrations of E2.
- Utilized immunocytochemical staining and antibody-based assays to detect DAT and ERs on cell membranes.
- Analyzed DAT levels in membrane and intracellular compartments using immunoblotting.
Main Results:
- Low concentrations of E2 rapidly inhibited DAT activity, suggesting a non-genomic mechanism.
- DAT and ERalpha levels increased after NGFbeta treatment; ERbeta was not elevated.
- DAT, ERalpha, and ERbeta were detected on the plasma membrane, with GPR30 also present.
- E2 treatment led to a decline in both membrane and intracellular DAT, without affecting total protein levels.
Conclusions:
- Physiological E2 levels may sequester DAT into intracellular compartments, affecting transporter function.
- Rapid estrogenic actions on neuronal cells are likely regulated by various membrane ERs.
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