Androgens differentially potentiate mouse intestinal smooth muscle by nongenomic activation of polyamine synthesis and Rho kinase activation.
We demonstrate that testosterone and its active metabolite 5alpha-dihydrotestosterone acutely (approximately 30 min) potentiate mouse ileal, but not duodenal, muscle activity. Androgens augment the amplitude of spontaneous peak-to-peak oscillations, alter the spontaneous activity frequency spectrum, and increase the amplitude of calcium-induced and carbachol-induced contractions. Concentration-dependence analyses revealed that maximal potentiation (449-910%) occurred at physiological concentrations of androgens (100 pM to 10 nM) with EC50 values in the picomolar range (8-20 pM). Western blot analyses using an antiandrogen receptor (anti-AR) antibody revealed the presence of two different AR proteins migrating at 87 and 110 kDa in ileal, but not duodenal, extracts. Androgen-induced potentiation was prevented by preincubation with AR antagonists flutamide or cyproterone acetate but was unaffected by pretreatment with cycloheximide plus actinomycin D, indicating that potentiation was mediated by ARs via a novel nongenomic mechanism. Androgen effects were mimicked by polyamines putrescine and spermine and were blocked by the ornithine decarboxylase and S-adenosyl-L-methionine decarboxylase inhibitors alpha-difluoromethylornithine and berenil, respectively. Accordingly, androgens increase alpha-difluoromethylornithine-sensitive ornithine-decarboxylase- mediated L-ornithine decarboxylation in ileal tissues within the same time course as isometric potentiation. Both putrescine and dihydrotestosterone induced Ca2+ sensitization of ionomycin-permeabilized ileal smooth muscle. Finally, inhibition of the Rho kinase (ROK) pathway with the specific inhibitor Y27632 completely prevented androgen-induced potentiation. In agreement, androgens elicited ROK-induced Ser19 phosphorylation of myosin light chain 2 in ileal muscle. These data indicate that androgens potentiate ileal contractile activity by an AR-dependent nongenomic mechanism involving intracellular polyamine signaling and Ca2+ sensitization via ROK activation.
González-Montelongo
Maria C
MC
Laboratorio de Fisiología Animal, Departamento de Biología Animal, Facultad de Biología, Universidad de La Laguna, 38206 Tenerife, Spain.
Marín
Raquel
R
Gómez
Tomás
T
Díaz
Mario
M
D058891:5-alpha Reductase Inhibitors; D059002:Androgen Receptor Antagonists; D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D015317:Biogenic Polyamines / Q000096:biosynthesis*; D002118:Calcium / Q000494:pharmacology; D002352:Carrier Proteins / Q000494:pharmacology; D013196:Dihydrotestosterone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000166:cytology / Q000187:drug effects*; D008297:Male; D051379:Mice; D008954:Models, Biological; D009119:Muscle Contraction / Q000187:drug effects*; D009130:Muscle, Smooth / Q000187:drug effects* / Q000378:metabolism; D011944:Receptors, Androgen / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013739:Testosterone / Q000494:pharmacology; D015533:Transcriptional Activation / Q000502:physiology; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
D058891:5-alpha Reductase Inhibitors; D059002:Androgen Receptor Antagonists; D000728:Androgens; D015317:Biogenic Polyamines; D002352:Carrier Proteins; D011944:Receptors, Androgen; C039317:ornithine decarboxylase antizyme inhibitor; D013196:Dihydrotestosterone; D013739:Testosterone; D020741:rho GTP-Binding Proteins; D002118:Calcium