Cited by 64Estrogen and related hormone effectsEffects and risks of endocrine disrupting chemicalsReceptor Mechanisms and Signaling

Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line

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Milanesi, Lorena, de Boland, Ana Russo, Boland, Ricardo·2008·Journal of Cellular Biochemistry

Abstract

The classical model of 17beta-estradiol action has been traditionally described to be mediated by the estrogen receptor (ER) localized exclusively in the nucleus. However, there is increasing functional evidence for extra nuclear localization of ER. We present biochemical, immunological and molecular data supporting mitochondrial-microsomal localization of ER alpha in the C2C12 skeletal muscle cell line. We first established [(3)H]17beta estradiol binding characteristics in whole cells in culture. Specific and saturable [(3)H]17beta estradiol binding sites of high affinity were then detected in mitochondrial fractions (K(d) = 0.43 nM; B(max) = 572 fmol/mg protein). Immunocytological studies revealed that estrogen receptors mainly localize at the mitochondrial and perinuclear level. These results were also confirmed using fluorescent 17beta estradiol-BSA conjugates. The immunoreactivity did not translocate into the nucleus by 17beta-estradiol treatment. Western and Ligand blot approaches corroborated the non-classical localization. Expression and subcellular distribution of ER alpha proteins were confirmed in C2C12 cells transfected with ER alpha siRNA and by RT-PCR employing specific primers. The non-classical distribution of native pools of ER alpha in skeletal muscle cells suggests an alternative mode of ER localization/function.

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  1. Drittanti L. 1988. Ph.D. thesis. ''Estudios sobre la accio ´n del 1,25(OH) 2 vitamina D 3 en el metabolismo de ce ´lulas musculares embrionarias cultivadas in vitro''. Universi- dad Nacional del Sur. Bahı ´a Blanca, Argentina.
  2. Jensen EV, Jacobson HI. 1960. Fate of steroidal estrogen in target tissues. In: Pincus G, Vollmer EP, editors. Bio- logical activities of steroids in relation to cancer. New York: Academic Press. pp 161-174.
  3. Vasconsuelo A, Milanesi L, Boland R. 2007. 17b-estradiol abrogates apoptosis in murine skeletal muscle cells through estrogen receptors: Role of the PI3K/Akt path- way. J Endrocrinol (in press).

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Authors
Milanesi, L., de Boland, A. R., & Boland, R.
Affiliations
Universidad Nacional del Sur (AR)
Corresponding author
Boland, Ricardo
Venue
Journal of Cellular Biochemistry
Publisher
Wiley
Published
2008-03-17
Volume
104
Issue
4
Pages
1254-1273
ISSN
  • 0730-2312 (ISSN-L)
  • 1097-4644
OpenAlex
W2007979384
Cited by
64
References
52
Citations by year
2025: 1 · 2024: 3 · 2023: 4 · 2022: 1 · 2021: 5 · 2020: 4 · 2019: 7 · 2018: 1 · 2017: 3 · 2016: 3 · 2015: 3 · 2014: 6 · 2013: 5 · 2012: 3
FWCI
2.85
Type
journal article
Language
en
Open access
Locations
  • Journal of Cellular Biochemistry (publishedVersion)
  • PubMed (publishedVersion)
  • CONICET Digital (CONICET) (submittedVersion, cc-by-nc-sa)
Funders
  • Consejo Nacional de Investigaciones Científicas y Técnicas
  • Agencia Nacional de Promoción Científica y Tecnológica (CONICET)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Effects and risks of endocrine disrupting chemicals
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
estrogen receptor alpha, mitochondrial localization, skeletal muscle cells, C2C12 myoblasts, subcellular localization, nuclear translocation, siRNA knockdown
MeSH terms
Animals, Cell Line, Estradiol, Female, Immunohistochemistry, Microsomes, Mitochondria, Muscle, Skeletal, Protein Transport, Estrogen Receptor alpha, Mice

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