Cloning and expression of a P2y purinoceptor from the adult bovine corpus callosum.
We have isolated an ATP receptor clone by screening a bovine corpus callosum cDNA library. The clone includes one open reading frame encoding for a protein of 373 amino acid residues (42 kDa) which belongs to the G-protein-coupled receptor superfamily. In Xenopus oocytes, this clone expressed an ATP receptor that triggered an oscillatory current in response to ATP (EC50 approximately 20 microM). This current may have resulted from the activation of phospholipase C, the formation of inositol trisphosphate, and the release of Ca2+, which then opens Cl- channels. The order of potency for ATP receptor agonists was 2-MeSATP approximately ATP >> alpha, beta-MeATP > adenosine, and UTP was ineffective, a pharmacological profile consistent with that of a P2y purinoceptor. Northern blot analysis of mRNAs from various bovine brain tissues showed that the gene is expressed in the cerebellum, medulla, corpus callosum, hippocampus, superior colliculus, frontal cortex, and retina. In situ RT-PCR showed transcripts of the gene in many glial cells and endothelial cells of the corpus callosum. The cloned receptor may play an important role in neuron-glial signaling under normal and pathological conditions.
Deng
G
G
Department of Psychobiology, University of California at Irvine 92697, USA.
Matute
C
C
Kumar
C K
CK
Fogarty
D J
DJ
Miledi
R
R
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D003337:Corpus Callosum / Q000737:chemistry / Q000378:metabolism*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000187:drug effects; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D012313:RNA / Q000008:administration & dosage / Q000235:genetics; D012315:RNA Caps / Q000235:genetics; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics; D018048:Receptors, Purinergic P2 / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014981:Xenopus
D018076:DNA, Complementary; D012315:RNA Caps; D012333:RNA, Messenger; D018048:Receptors, Purinergic P2; D012313:RNA; D000255:Adenosine Triphosphate