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Fluorescently-Labeled Estradiol Internalization and Membrane Trafficking in Live N-38 Neuronal Cells Visualized with Total Internal Reflection Fluorescence Microscopy
Citation
Kisler, K and Chow, RH and Dominguez, R, Fluorescently-Labeled Estradiol Internalization and Membrane Trafficking in Live N-38 Neuronal Cells Visualized with Total Internal Reflection Fluorescence Microscopy, Journal of steroids & hormonal science, Suppl 12 pp. 1-15. ISSN 2157-7536 (2013) [Refereed Article]
Copyright Statement
Copyright: © 2013 Kisler K, et al. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/
DOI: doi:10.4172/2157-7536.S12-002
Abstract
Estradiol is a steroid hormone that binds and activates estradiol receptors. Activation of these
receptors is known to modulate neuronal physiology and provide neuroprotection, but it is not
completely understood how estradiol mediates these actions on the nervous system. Activation of
a sub-population of estradiol receptor-α (ERα), originally identified as a nuclear protein, localizes
to the plasma membrane and appears to be a critical step in neuroprotection against brain injury
and disease. Previously we showed that estradiol stimulates the rapid and transient trafficking of
plasma membrane ERα in primary hypothalamic neurons, and internalization of membraneimpermeant
estradiol (E6BSA-FITC) into cortical neuron endosomes in vitro. These findings
support the concept that estradiol activates and down-regulates plasma membrane ERα by
triggering endocytosis. Here, we use TIRFM (total internal reflection fluorescence microscopy) to
image the trafficking of E6BSA-FITC, and GFP-labeled ERα, in live cells in real time. We show
that activation of plasma membrane ERs by E6BSA-FITC result in internalization of the
fluorescent ligand in live N-38 neurons, an immortalized hypothalamic cell line. Pretreatment with
ER antagonist ICI 182,780 decreased the number of E6BSA-FITC labeled puncta observed. We
also observed in live N-38 neurons that E6BSA-FITC co-localized with FM4-64 and LysoTracker
fluorescent dyes that label endosomes and lysosomes. Our results provide further evidence that
plasma membrane ERα activation results in endocytosis of the receptor.
Item Details
| Item Type: | Refereed Article |
|---|---|
| Keywords: | Live cell imaging, TIRF, trafficking, estrogen receptor |
| Research Division: | Biological Sciences |
| Research Group: | Zoology |
| Research Field: | Animal Cell and Molecular Biology |
| Objective Division: | Expanding Knowledge |
| Objective Group: | Expanding Knowledge |
| Objective Field: | Expanding Knowledge in the Biological Sciences |
| Author: | Dominguez, R (Dr Reymundo Dominguez) |
| ID Code: | 122240 |
| Year Published: | 2013 |
| Deposited By: | Medicine (Discipline) |
| Deposited On: | 2017-11-06 |
| Last Modified: | 2017-11-20 |
| Downloads: | 41 View Download Statistics |
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