title: P2X2 subunits contribute to fast synaptic excitation in myenteric neurons of the mouse small intestine.
abstract: P2X receptors are ATP-gated cation channels composed of one or more of seven different subunits. ATP acts at P2X receptors to contribute to fast excitatory postsynaptic potentials (fEPSPs) in myenteric neurons but the subunit composition of enteric P2X receptors is unknown. These studies used tissues from P2X2 wild-type (P2X2+/+) and P2X2 gene knockout (P2X2-/-) mice to investigate the role of this subunit in enteric neurotransmission. Intracellular electrophysiological methods were used to record synaptic and drug-induced responses from ileal myenteric neurons in vitro. Drug-induced longitudinal muscle contractions and peristaltic contractions of ileal segments were also studied in vitro. Gastrointestinal transit was measured as the progression in 30 min of a liquid radioactive marker administered by gavage to fasted mice. RT-PCR analysis of mRNA from intestinal tissues and data from immunohistochemical studies verified P2X2 gene deletion. The fEPSPs recorded from S neurons in tissues from P2X2+/+ mice were reduced by mecamylamine (nicotinic cholinergic receptor antagonist) and PPADS (P2X receptor antagonist). The fEPSPs recorded from S neurons from P2X2-/- mice were unaffected by PPADS but were blocked by mecamylamine. ATP depolarized S and AH neurons from P2X2+/+ mice. ATP depolarized AH but not S neurons from P2X2-/- mice. alpha,beta-Methylene ATP (alpha,beta-mATP)(an agonist at P2X3 subunit-containing receptors) did not depolarize S neurons but it did depolarize AH neurons in P2X2+/+ and P2X2-/- mice. Peristalsis was inhibited in ileal segments from P2X2-/- mice but longitudinal muscle contractions caused by nicotine and bethanechol were similar in segments from P2X2+/+ and P2X2-/- mice. Gastrointestinal transit was similar in P2X2+/+ and P2X2-/- mice. It is concluded that P2X2 homomeric receptors contribute to fEPSPs in neural pathways underlying peristalsis studied in vitro.
authors.affiliation: Department of Pharmacology and Toxicology and the Neuroscience Program, Michigan State University, East Lansing, MI 48824, USA.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Ren
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Bian
authors.forename: Matthew
authors.initials: M
authors.lastname: DeVries
authors.forename: Birthe
authors.initials: B
authors.lastname: Schnegelsberg
authors.forename: Debra A
authors.initials: DA
authors.lastname: Cockayne
authors.forename: Anthony P D W
authors.initials: AP
authors.lastname: Ford
authors.forename: James J
authors.initials: JJ
authors.lastname: Galligan
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D004594:Electrophysiology; D005769:Gastrointestinal Motility; D005772:Gastrointestinal Transit / Q000502:physiology; D017353:Gene Deletion; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007421:Intestine, Small / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D009197:Myenteric Plexus / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D010528:Peristalsis / Q000502:physiology; D020033:Protein Isoforms / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D011930:Reaction Time; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000502:physiology*; D058476:Receptors, Purinergic P2X2; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013569:Synapses / Q000502:physiology*; D014018:Tissue Distribution
chemical_list: C546402:P2rx2 protein, mouse; D020033:Protein Isoforms; D012333:RNA, Messenger; D018048:Receptors, Purinergic P2; D058476:Receptors, Purinergic P2X2; D000255:Adenosine Triphosphate; C002630:alpha,beta-methyleneadenosine 5'-triphosphate