Perimembrane localization of the estrogen receptor α protein in neuronal processes of cultured hippocampal neurons

Charlotte H. Clarke, Andrea M. Norfleet, Mark S F Clarke, Cheryl S. Watson, Kathryn Cunningham, Mary L. Thomas

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

There is clear evidence of rapid, nongenomic responses to estrogen in a variety of neruronal model systems. To address the question of whether some of these rapid estrogen signals might be transduced by the classical estrogen receptor (ER) α or a closely related protein in nontransformed neurons, we undertook the present study using isolated fetal rat hippocampal neurons. Several antibodies developed to detect ERα were tested in this system and showed positive membrane staining in nonpermeabilized neurons. MC-20, an affinity purified anti-ERα, rabbit polyclonal IgG antibody which does not recognize ERβ was selected to carry out the majority of the experiments. When permeabilized, the hippocampal neurons exhibited low levels of nuclear staining for ERα, but abundant labeling for ERα throughout the entire cell including the neurites. In addition to traditional immunocytochemistry controls, incubation of neurons for 24 h in the presence of 10 μM antisense oligonucleotide directed against the translation start site of ERα reduced ERα immunoreactivity throughout the neurons providing further evidence that the immunostaining was specific for ERα. Confocal and conventional microscopy demonstrated that the antigen was predominately extranuclear and localization of ERα in the neurites suggests that the receptor is in close proximity to the plasma membrane. This localization is consistent with a role for ERα as a transducer of rapid, nongenomic estrogen responses in hippocampal neurons.

Original languageEnglish
Pages (from-to)34-42
Number of pages9
JournalNeuroendocrinology
Volume71
Issue number1
StatePublished - 2000

Fingerprint

Estrogen Receptors
Neurons
Estrogens
Neurites
estrophilin
Staining and Labeling
Antibodies
Antisense Oligonucleotides
Transducers
Confocal Microscopy
Immunoglobulin G
Immunohistochemistry
Cell Membrane
Rabbits
Antigens
Membranes

Keywords

  • Gonadal steroid receptors
  • Hippocampal neurons
  • Immunocytochemistry
  • Membrane steroid receptors

ASJC Scopus subject areas

  • Endocrinology
  • Neuroscience(all)

Cite this

Clarke, C. H., Norfleet, A. M., Clarke, M. S. F., Watson, C. S., Cunningham, K., & Thomas, M. L. (2000). Perimembrane localization of the estrogen receptor α protein in neuronal processes of cultured hippocampal neurons. Neuroendocrinology, 71(1), 34-42.

Perimembrane localization of the estrogen receptor α protein in neuronal processes of cultured hippocampal neurons. / Clarke, Charlotte H.; Norfleet, Andrea M.; Clarke, Mark S F; Watson, Cheryl S.; Cunningham, Kathryn; Thomas, Mary L.

In: Neuroendocrinology, Vol. 71, No. 1, 2000, p. 34-42.

Research output: Contribution to journalArticle

Clarke, CH, Norfleet, AM, Clarke, MSF, Watson, CS, Cunningham, K & Thomas, ML 2000, 'Perimembrane localization of the estrogen receptor α protein in neuronal processes of cultured hippocampal neurons', Neuroendocrinology, vol. 71, no. 1, pp. 34-42.
Clarke, Charlotte H. ; Norfleet, Andrea M. ; Clarke, Mark S F ; Watson, Cheryl S. ; Cunningham, Kathryn ; Thomas, Mary L. / Perimembrane localization of the estrogen receptor α protein in neuronal processes of cultured hippocampal neurons. In: Neuroendocrinology. 2000 ; Vol. 71, No. 1. pp. 34-42.
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