title: Localization and functional expression of splice variants of the P2X2 receptor.
abstract: cDNAs encoding three splice variants of the P2X2 receptor were isolated from rat cerebellum. The first variant has a serine/proline-rich segment deleted from the intracellularly located carboxyl-terminal domain of the P2X2 subunit. The second and third variants have the splice site in the second half of the predicted first transmembrane domain. Either a 12-amino acid insertion or a six-amino acid deletion occurs at this position. cRNAs for these isoforms of the P2X2 subunit were injected into Xenopus laevis oocytes and tested for function. ATP evoked inward currents only with the splice variant [designated P2X2(b)] having the 69-amino acid deletion. The potencies of various agonists at the homomeric P2X2(b) receptor were not significantly different from those at the P2X2(a) homomeric channel. However, the P2X2(b) receptor showed significantly lower antagonist sensitivity. In contrast to the nondesensitizing P2X2(a) receptor, prolonged application of ATP produced a more rapid desensitization of the P2X2(b) receptor. When the P2X2(a) and P2X2(b) receptor responses were recorded in transfected mammalian cells, this difference was again found. The change in desensitization may be determined by proline/serine-rich segments and/or phosphorylation motifs that are removed from the tail region in formation of the P2X2(b) subunit. In situ hybridization of the three newly isolated isoforms of the P2X2 subunit was performed at the macroscopic and cellular levels; transcripts for two of them [P2X2(b) and p2x2(c)] but not the third [p2x2(d)], which carries the 12-amino acid addition, were present in many structures in the neonatal rat brain and on sensory and sympathetic ganglia. mRNA for the p2x2(d) splice variant was present only in the nodose ganglion, at a low level.
authors.affiliation: Molecular Neurobiology Unit, Royal Free Hospital School of Medicine, London, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: Simon
authors.forename: E J
authors.initials: EJ
authors.lastname: Kidd
authors.forename: F M
authors.initials: FM
authors.lastname: Smith
authors.forename: I P
authors.initials: IP
authors.lastname: Chessell
authors.forename: R
authors.initials: R
authors.lastname: Murrell-Lagnado
authors.forename: P P
authors.initials: PP
authors.lastname: Humphrey
authors.forename: E A
authors.initials: EA
authors.lastname: Barnard
mesh_terms: D000255:Adenosine Triphosphate / Q000502:physiology; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D002451:Cell Compartmentation; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004553:Electric Conductivity; D006801:Humans; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018048:Receptors, Purinergic P2 / Q000235:genetics*; D058476:Receptors, Purinergic P2X2; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: D018076:DNA, Complementary; C546401:P2RX2 protein, human; C546403:P2rx2 protein, rat; D012333:RNA, Messenger; D018048:Receptors, Purinergic P2; D058476:Receptors, Purinergic P2X2; D000255:Adenosine Triphosphate