α1-adrenoceptor activation induces phosphorylation of β2-adrenoceptors in human prostate tissue

Authors


Christian Gratzke, Urologische Klinik, Universitäts-Klinikum Großhadern, Marchioninistr. 15, 81377 Munich, Germany. e-mail: christian.gratzke@med.uni-muenchen.de

Abstract

OBJECTIVE

  • • To test whether β1-adrenoceptor activation leads to phosphorylation of the β2-adrenoceptor in human prostate tissue.

PATIENTS AND METHODS

  • • Prostate tissue from patients undergoing radical prostatectomy was stimulated in vitro with the α1-adrenergic agonist phenylephrine (10 µM).
  • • α2-adrenoceptor phosphorylation at serines 345/346 was studied using Western blot analysis with a phospho-specific antibody.
  • • The role of second messenger kinases was assessed by studying the effects of the protein kinase C (PKC) inhibitor Ro 31-8425 and the protein kinase A (PKA) inhibitor H89 on phenylephrine-induced phosphorylation.
  • • The expression of G protein-coupled receptor kinases (GRKs) 2/3 was analysed using quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR), Western blot analysis and immunohistochemistry.

RESULTS

  • • Stimulation of prostate tissue with phenylephrine resulted in phosphorylation of the β2-adrenoceptor (5, 10 and 20 min after stimulation).
  • • This α1-adrenoceptor-induced phosphorylation of β2-adrenoceptors was resistant to inhibition of PKC and PKA.
  • • Changes in phosphorylation levels were not attributable to changes in receptor levels, as these remained constant during stimulation.
  • • RT-PCR and Western blot analysis showed expression of GRK2/3 in human prostate tissues.
  • • Immunohistochemical staining showed that GRK2/3 expression in human prostate tissue is located to stromal and smooth muscle cells.

CONCLUSIONS

  • • Activation of α1-adrenoceptors causes phosphorylation of β2-adrenoceptors in the human prostate. This may enhance α1-adrenergic contraction and is possibly mediated by GRK2, which is expressed in prostate smooth muscle.
  • • Mutual regulation between different adrenergic receptors might be involved in the therapeutic effects of α1-blockers in patients with benign prostate hyperplasia.

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