title: Immunofluorescence analysis of antisense oligodeoxynucleotide-mediated 'knock-down' of the mouse delta opioid receptor in vitro and in vivo.
abstract: We have previously used antisense oligodeoxynucleotides (ODN) to the cloned delta opioid receptor (DOR) to inhibit the antinociceptive response to spinally administered delta opioid receptor selective agonists in mice. Here we have examined the effect of DOR antisense ODN treatment on the level of DOR expressed in NG 108-15 cells and the spinal cord, through immuno-fluorescence microscopy, to determine the efficiency and selectivity of the antisense ODN-mediated "knock-down' of the DOR in these tissues. Antisense ODN, but not mismatch control, treatment resulted in a significant reduction in DOR immunoreactivity (-ir) in NG 108-15 cells and spinal cord. Thus, the inhibition of antinociceptive response to intrathecal delta selective agonists by DOR antisense ODN correlates with the loss of DOR-ir in the superficial layers of the dorsal horn of the spinal cord.
authors.affiliation: Department of Pharmacology, University of Arizona Health Sciences Center, Tucson 85724, USA. lai@aruba.ccit.arizona.edu
authors.forename: J
authors.initials: J
authors.lastname: Lai
authors.forename: M
authors.initials: M
authors.lastname: Riedl
authors.forename: L S
authors.initials: LS
authors.lastname: Stone
authors.forename: U
authors.initials: U
authors.lastname: Arvidsson
authors.forename: E J
authors.initials: EJ
authors.lastname: Bilsky
authors.forename: G L
authors.initials: GL
authors.lastname: Wilcox
authors.forename: R
authors.initials: R
authors.lastname: Elde
authors.forename: F
authors.initials: F
authors.lastname: Porreca
mesh_terms: D000818:Animals; D016375:Antisense Elements (Genetics) / Q000493:pharmacokinetics*; D001692:Biological Transport / Q000502:physiology; D002460:Cell Line / Q000737:chemistry; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D007278:Injections, Spinal; D051379:Mice; D009619:Nociceptors / Q000502:physiology; D015345:Oligonucleotide Probes / Q000493:pharmacokinetics*; D017465:Receptors, Opioid, delta / Q000032:analysis* / Q000235:genetics*; D013116:Spinal Cord / Q000737:chemistry; D014966:Xanthenes
chemical_list: D016375:Antisense Elements (Genetics); D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D017465:Receptors, Opioid, delta; D014966:Xanthenes; C034657:Texas red