Differential expression of P2X-purinoceptor subtypes in circular and longitudinal muscle of canine colon.
Adenosine triphosphate (ATP) mediates excitatory junction potentials through P2X receptors in many smooth muscles. However, relatively little is known about postjunctional intestinal P2X receptors. We examined the effect of exogenous ATP on circular and longitudinal myocytes of canine colon using the patch clamp technique at 32 degrees C. In both cell types, ATP induced inward currents (I(ATP)) at -70 mV in a concentration-dependent manner. The potency profile of ATP analogues in circular myocytes was: ATP approximately 2-methylthio-ATP > alpha,beta-methylene ATP, and that in longitudinal myocytes was: alpha,beta-methylene ATP approximately ATP > 2-methylthio-ATP. Pretreatment of circular myocytes with alpha,beta-methylene ATP inhibited the response to subsequent ATP, suggesting receptor desensitization. I-V relationships of I(ATP) were linear with inward rectification and E(rev) of -13 mV. I(ATP) at -70 mV was carried predominantly by Na+ as determined by shifts in E(rev) when extracellular Na+ was lowered. In RT-PCR, circular myocytes expressed mRNAs encoding P2X2, 3 and 4, while longitudinal myocytes expressed mRNAs for P2X3 and 5. P2X7 was absent in both cells. Fragments of each subtype were cloned and sequenced. We failed to clone P2X1 and P2X6 genes. Overall, different P2X receptor subtypes are expressed in circular and longitudinal canine colonic myocytes. Their activation produces non-selective cation currents that can depolarize and excite muscles of both layers.
Lee
H K
HK
Department of Pharmacology, University of Ulsan College of Medicine, Seoul, Korea. kyung@amc.seoul.kr
Ro
S
S
Keef
K D
KD
Kathy
K D
KD
Kim
Y-H
YH
Kim
H W
HW
Horowitz
B
B
Sanders
K M
KM
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000502:physiology; D003106:Colon / Q000502:physiology*; D004285:Dogs; D006801:Humans; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D009119:Muscle Contraction / Q000502:physiology; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D018408:Patch-Clamp Techniques; D020033:Protein Isoforms / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017385:Sequence Homology
D020033:Protein Isoforms; D012333:RNA, Messenger; D018048:Receptors, Purinergic P2; D000255:Adenosine Triphosphate