Cloning and Characterization of a Mouse σ1 Receptor |
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Authors: | Ying-Xian Pan Jianfeng Mei Jin Xu Ban-Lin Wan Amy Zuckerman Gavril W Pasternak |
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Institution: | Cotzias Laboratory of Neuro-Oncology, Memorial Sloan-Kettering Cancer Center, New York, New York, U.S.A. |
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Abstract: | Abstract: A cDNA clone (S2-1a) isolated from a mouse brain cDNA library, using a guinea pig σ1 cDNA as probe, has high homology to the predicted protein sequence of the guinea pig (88%) and human (90%) σ1 receptors. Northern analysis revealed a major mRNA of ~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 σ1 receptors. In vitro translation yielded a protein of ~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 σ1 receptor. An antisense oligodeoxynucleotide designed from S2-1a potentiated opioid analgesia in vivo. |
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Keywords: | Opioid Analgesia Antiopioid Cloning Antisense σ receptor Pentazocine |
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