title: P2Y1 purinoreceptors are fundamental to inhibitory motor control of murine colonic excitability and transit.
abstract: Activation of enteric inhibitory motor neurons causes inhibitory junctional potentials (IJPs) and muscle relaxation in mammalian gastrointestinal (GI) muscles, including humans. IJPs in many GI muscles are bi-phasic with a fast initial hyperpolarization (fIJP) due to release of a purine neurotransmitter and a slower hyperpolarization component (sIJP) due to release of nitric oxide. We sought to characterize the nature of the post-junctional receptor(s) involved in transducing purinergic neural inputs in the murine colon using mice with genetically deactivated P2ry1. Wild-type mice had characteristic biphasic IJPs and pharmacological dissection confirmed that the fIJP was purinergic and the sIJP was nitrergic. The fIJP was completely absent in P2ry1(−/−) mice and the P2Y1 receptor antagonist MRS2500 had no effect on electrical activity or responses to electrical field stimulation of intrinsic nerves in these mice. Contractile experiments confirmed that purinergic responses were abolished in P2ry1(−/−) mice. Picospritzing of neurotransmitter candidates (ATP and its primary metabolite, ADP) and β-NAD (and its primary metabolite, ADP-ribose, ADPR) caused transient hyperpolarization responses in wild-type colons, but responses to β-NAD and ADPR were completely abolished in P2ry1(−/−) mice. Hyperpolarization and relaxation responses to ATP and ADP were retained in colons of P2ry1(−/−) mice. Video imaging revealed that transit of fecal pellets was significantly delayed in colons from P2ry1(−/−) mice. These data demonstrate the importance of purinergic neurotransmission in regulating colonic motility and confirm pharmacological experiments suggesting that purinergic neurotransmission is mediated via P2Y1 receptors.
authors.affiliation: Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV 89557, USA.
authors.forename: Sung Jin
authors.initials: SJ
authors.lastname: Hwang
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Blair
authors.forename: Leonie
authors.initials: L
authors.lastname: Durnin
authors.forename: Violeta
authors.initials: V
authors.lastname: Mutafova-Yambolieva
authors.forename: Kenton M
authors.initials: KM
authors.lastname: Sanders
authors.forename: Sean M
authors.initials: SM
authors.lastname: Ward
mesh_terms: D000818:Animals; D003106:Colon / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D004558:Electric Stimulation / Q000379:methods; D005769:Gastrointestinal Motility / Q000187:drug effects / Q000502:physiology; D005772:Gastrointestinal Transit / Q000187:drug effects / Q000502:physiology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism*; D009126:Muscle Relaxation / Q000187:drug effects / Q000502:physiology; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D009469:Neuromuscular Junction / Q000187:drug effects / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D058919:Purinergic P2 Receptor Antagonists / Q000494:pharmacology; D011687:Purines / Q000378:metabolism; D058472:Receptors, Purinergic P2Y1 / Q000378:metabolism*; D009435:Synaptic Transmission / Q000187:drug effects
chemical_list: D018377:Neurotransmitter Agents; D058919:Purinergic P2 Receptor Antagonists; D011687:Purines; D058472:Receptors, Purinergic P2Y1; D009569:Nitric Oxide; C030985:purine