title: Cloning and characterization of a mouse sigma1 receptor.
abstract: A cDNA clone (S2-1a) isolated from a mouse brain cDNA library, using a guinea pig sigma1 cDNA as probe, has high homology to the predicted protein sequence of the guinea pig (88%) and human (90%) sigma1 receptors. Northern analysis revealed a major mRNA of approximately 1.8 kb in a wide range of mouse tissues, with highest levels in brain, liver, kidney, and thymus. Southern analysis and chromosomal mapping in the mouse suggested a single-copy gene in region A5-B2 of chromosome 4. Expression of the clone in MCF-7 and CHO cells led to a pronounced increase in (+)-[3H]pentazocine binding with a selectivity profile consistent with sigma1 receptors. In vitro translation yielded a protein of approximately 28 kDa, as did transfection of a probe containing the hemagglutinin (HA) epitope (S2-1a.HA) into CHO cells, as determined by western analysis using an antibody directed against HA. (+)-[3H]-Pentazocine binding to immunopurified HA-tagged receptor demonstrated conclusively that S2-1a.HA encodes a high-affinity (+)-[3H]pentazocine binding site with characteristics of a murine sigma1 receptor. An antisense oligodeoxynucleotide designed from S2-1a potentiated opioid analgesia in vivo.
authors.affiliation: Cotzias Laboratory of Neuro-Oncology, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
authors.forename: Y X
authors.initials: YX
authors.lastname: Pan
authors.forename: J
authors.initials: J
authors.lastname: Mei
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: B L
authors.initials: BL
authors.lastname: Wan
authors.forename: A
authors.initials: A
authors.lastname: Zuckerman
authors.forename: G W
authors.initials: GW
authors.lastname: Pasternak
mesh_terms: D000595:Amino Acid Sequence; D000701:Analgesics, Opioid / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D006224:Cricetinae; D015870:Gene Expression; D015723:Gene Library; D006168:Guinea Pigs; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009292:Narcotic Antagonists / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D009928:Organ Specificity; D010147:Pain Measurement; D010423:Pentazocine / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D000701:Analgesics, Opioid; D009292:Narcotic Antagonists; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D010423:Pentazocine