Extracellular UDP enhances P2X-mediated bladder smooth muscle contractility via P2Y(6) activation of the phospholipase C/inositol trisphosphate pathway.
Bladder dysfunction characterized by abnormal bladder smooth muscle (BSM) contractions is pivotal to the disease process in overactive bladder, urge incontinence, and spinal cord injury. Purinergic signaling comprises one key pathway in modulating BSM contractility, but molecular mechanisms remain unclear. Here we demonstrate, using myography, that activation of P2Y6 by either UDP or a specific agonist (MRS 2693) induced a sustained increase in BSM tone (up to 2 mN) in a concentration-dependent manner. Notably, activation of P2Y6 enhanced ATP-mediated BSM contractile force by up to 45%, indicating synergistic interactions between P2X and P2Y signaling. P2Y6-activated responses were abolished by phospholipase C (PLC) and inositol trisphosphate (IP3) receptor antagonists U73122 and xestospongin C, demonstrating involvement of the PLC/IP3 signal pathway. Mice null for Entpd1, an ectonucleotidase on BSM, demonstrated increased force generation on P2Y6 activation (150%). Thus, in vivo perturbations to purinergic signaling resulted in altered P2Y6 activity and bladder contractility. We conclude that UDP, acting on P2Y6, regulates BSM tone and in doing so selectively maximizes P2X1-mediated contraction forces. This novel neurotransmitter pathway may play an important role in urinary voiding disorders characterized by abnormal bladder motility.
Laboratory of Voiding Dysfunction, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
Weiqun
W
Yu
Xiaofeng
X
Sun
Simon C
SC
Robson
Warren G
WG
Hill
D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D001081:Apyrase / Q000235:genetics; D004963:Estrenes / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D051379:Mice; D009119:Muscle Contraction / Q000187:drug effects*; D011760:Pyrrolidinones / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D058474:Receptors, Purinergic P2X1 / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014530:Uridine Diphosphate / Q000494:pharmacology; D001743:Urinary Bladder / Q000187:drug effects
D015703:Antigens, CD; D004963:Estrenes; D011760:Pyrrolidinones; D018048:Receptors, Purinergic P2; D058474:Receptors, Purinergic P2X1; C099845:purinoceptor P2Y6; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D014530:Uridine Diphosphate; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D010738:Type C Phospholipases; D001081:Apyrase; C067869:CD39 antigen