Cell and Tissue Research

, Volume 342, Issue 2, pp 295–306

β−Adrenergic receptor subtype expression in myocyte and non-myocyte cells in human female bladder

Authors

    • Women’s Health New Business DevelopmentProcter & Gamble Pharmaceuticals now Warner Chilcott Pharmaceuticals Inc.
    • The Procter & Gamble Company
  • Karl-Erik Andersson
    • Wake Forest Institute for Regenerative MedicineWake Forest University School of Medicine
  • F. Aura Kullmann
    • Department of Pharmacology and Chemical BiologyUniversity of Pittsburgh
    • Urogenix Inc.
  • Glenna Burmer
    • LifeSpan BioSciences, Inc.
  • William C. de Groat
    • Department of Pharmacology and Chemical BiologyUniversity of Pittsburgh
  • Jan S. Rosenbaum
    • Women’s Health New Business DevelopmentProcter & Gamble Pharmaceuticals now Warner Chilcott Pharmaceuticals Inc.
    • CincyTech USA
Regular Article

DOI: 10.1007/s00441-010-1053-x

Cite this article as:
Limberg, B.J., Andersson, K., Aura Kullmann, F. et al. Cell Tissue Res (2010) 342: 295. doi:10.1007/s00441-010-1053-x

Abstract

β3-Adrenergic receptor agonists are currently under clinical development for the treatment of overactive bladder, a condition that is prevalent in postmenopausal women. These agents purportedly relax bladder smooth muscle through a direct action at the myocyte β3-receptor. The aim of this study was to examine the expression of the individual beta-adrenergic receptors in full thickness sections from ageing human female bladder. We obtained a series of rabbit polyclonal antibodies generated against each of the three β-adrenergic receptors, and validated their receptor specificity in CHOK1 cells expressing each of the individual receptors. Immunostaining for β1, β2, and β3 were each more prominent in the urothelium than in the detrusor, with all receptors expressed in the same cell types, indicating co-expression of all three receptors throughout the urothelium in addition to the detrusor. Staining of all receptors was also observed in suburothelial myofibroblast-like cells, intramural ganglion cells, and in Schwann cells of intramural nerves. The β3-receptor in the human urothelium appears to be functional, as two different selective β3-receptor agonists, TAK677 and BRL37344, stimulate cAMP formation in UROtsa cells. Densitometry analysis indicates a persistent expression of all receptors throughout the bladder with increasing age, with the exception of the β2-receptor in the urothelium of the trigone, which appears to decrease slightly in older women. These data indicate that β3-receptor expression is maintained with age, but may function in concert with other β-receptors. Activation of the myocyte receptor may be influenced by action on non-myocyte structures including the intramural ganglion cells and myofibroblasts.

Keywords

Overactive bladderβ-adrenergic receptorsImmunohistochemistryUrotheliumAge

Supplementary material

441_2010_1053_Fig9_ESM.gif (179 kb)
Supplemental Figure 1

Validation of specific staining for the human β3-AR with the KG115 antibody. The antibody was generated against the human β3-AR -COOH terminus. The antibody was used at a concentration of 10 μg/ml. Staining (red) were observed in transiently transfected CHOK1 cells with the human β2-AR (b) and β3-AR (c) but not β1-AR (a) and empty vector (d). This suggests that the KG115 does not specifically recognize β3-AR by immunostaining. (GIF 179 kb)

441_2010_1053_MOESM1_ESM.tif (3 mb)
High Resolution Image (TIFF 3062 kb)
441_2010_1053_Fig10_ESM.gif (4 kb)
Supplemental Figure 1

Validation of specific staining for the human β3-AR with the KG115 antibody. The antibody was generated against the human β3-AR -COOH terminus. The antibody was used at a concentration of 10 μg/ml. Staining (red) were observed in transiently transfected CHOK1 cells with the human β2-AR (b) and β3-AR (c) but not β1-AR (a) and empty vector (d). This suggests that the KG115 does not specifically recognize β3-AR by immunostaining. (GIF 179 kb)

441_2010_1053_MOESM2_ESM.tif (388 kb)
High Resolution Image (TIFF 387 kb)

Copyright information

© Springer-Verlag 2010