Anatomical location and redistribution of G protein-coupled estrogen receptor-1 during the estrus cycle in mouse kidney and specific binding to estrogens but not aldosterone

Mol Cell Endocrinol. 2014 Feb 15;382(2):950-9. doi: 10.1016/j.mce.2013.11.005. Epub 2013 Nov 13.


Prior studies have linked renoprotective effects of estrogens to G-protein-coupled estrogen receptor-1 (GPER-1) and suggest that aldosterone may also activate GPER-1. Here, the role of GPER-1 in murine renal tissue was further evaluated by examining its anatomical distribution, subcellular distribution and steroid binding specificity. Dual immunofluorescent staining using position-specific markers showed that GPER-1 immunoreactivity primarily resides in distal convoluted tubules and the Loop of Henle (stained with Tamm-Horsfall Protein-1). Lower GPER-1 expression was observed in proximal convoluted tubules marked with megalin, and GPER-1 was not detected in collecting ducts. Plasma membrane fractions prepared from whole kidney tissue or HEK293 cells expressing recombinant human GPER-1 (HEK-GPER-1) displayed high-affinity, specific [(3)H]-17β-estradiol ([(3)H]-E2) binding, but no specific [(3)H]-aldosterone binding. In contrast, cytosolic preparations exhibited specific binding to [(3)H]-aldosterone but not to [(3)H]-E2, consistent with the subcellular distribution of GPER-1 and mineralocorticoid receptor (MR) in these preparations. Aldosterone and MR antagonists, spironolactone and eplerenone, failed to compete for specific [(3)H]-E2 binding to membranes of HEK-GPER-1 cells. Furthermore, aldosterone did not increase [(35)S]-GTP-γS binding to membranes of HEK-GPER-1 cells, indicating that it is not involved in G protein signaling mediated through GPER-1. During the secretory phases of the estrus cycle, GPER-1 is upregulated on cortical epithelia and localized to the basolateral surface during proestrus and redistributed intracellularly during estrus. GPER-1 is down-modulated during luteal phases of the estrus cycle with significantly less receptor on the surface of renal epithelia. Our results demonstrate that GPER-1 is associated with specific estrogen binding and not aldosterone binding and that GPER-1 expression is modulated during the estrus cycle which may suggest a physiological role for GPER-1 in the kidney during reproduction.

Keywords: Aldosterone binding; Distal convoluted tubules; Estradiol binding; Estrus cycle; GPER-1; Mineralocorticoid receptor.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aldosterone / metabolism
  • Animals
  • Eplerenone
  • Estradiol / metabolism*
  • Estrus / physiology*
  • Female
  • Gene Expression Regulation
  • Guanosine 5'-O-(3-Thiotriphosphate) / metabolism
  • HEK293 Cells
  • Humans
  • Kidney Tubules, Distal / cytology
  • Kidney Tubules, Distal / metabolism*
  • Loop of Henle / cytology
  • Loop of Henle / metabolism*
  • Low Density Lipoprotein Receptor-Related Protein-2 / genetics
  • Low Density Lipoprotein Receptor-Related Protein-2 / metabolism
  • Mice
  • Mineralocorticoid Receptor Antagonists / pharmacology
  • Protein Binding
  • Receptors, Estrogen
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism
  • Receptors, Mineralocorticoid / genetics
  • Receptors, Mineralocorticoid / metabolism
  • Reproduction / physiology*
  • Spironolactone / analogs & derivatives
  • Spironolactone / pharmacology


  • GPER1 protein, mouse
  • Low Density Lipoprotein Receptor-Related Protein-2
  • Lrp2 protein, mouse
  • Mineralocorticoid Receptor Antagonists
  • Receptors, Estrogen
  • Receptors, G-Protein-Coupled
  • Receptors, Mineralocorticoid
  • Spironolactone
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Aldosterone
  • Estradiol
  • Eplerenone