Antisense oligodeoxynucleotides to the cloned δ receptor DOR‐1: Uptake, stability, and regulation of gene expression Journal Article
| Authors: | Standifer, K. M.; Jenab, S.; Su, W.; Chien, C. C.; Pan, Y. X.; Inturrisi, C. E.; Pasternak, G. W. |
| Article Title: | Antisense oligodeoxynucleotides to the cloned δ receptor DOR‐1: Uptake, stability, and regulation of gene expression |
| Abstract: | Abstract: Phosphodiester antisense oligodeoxynucleotides (ODNs) directed against various domains of the cloned mouse δ receptor DOR‐1 reduce δ‐opioid receptor binding in vivo and in vitro. The present study examines the stability of an antisense ODN (275 nM) directed against the δ‐opioid receptor and its effect on DOR‐1 mRNA in cultured neuroblastoma cells and in vivo. When added to NG108‐15 cells, much of the antisense ODN is degraded. However, >1% is intact, associated with cells, and stable for at least 72 h. Northern blot analysis demonstrates that treatment of NG108‐15 cells with the antisense ODN reduces the levels of a species of DOR‐1 mRNA by ∼25%. Similarly, intrathecal administration of the antisense ODN results in the accumulation of intact ODN within the spinal cord, which is stable for at least 72 h, although the levels of accumulation in vivo are lower than in vitro after either 4 or 72 h. Antisense ODN treatment lowers DOR‐1 mRNA levels by ∼25%. The loss of mRNA both in vivo and in vitro corresponds quite well to the decreases in receptor binding previously observed by our laboratory and is consistent with reduction of δ‐opioid receptor protein in vitro as determined by western blot with a monoclonal antibody selective for the δ‐opioid receptor. In conclusion, these studies indicate that a small, but significant, proportion of ODN is taken up by cells and remains intact for up to 72 h. This appears to be sufficient to down‐regulate mRNA levels of δ‐opioid receptors and their expression. Copyright © 1995, Wiley Blackwell. All rights reserved |
| Keywords: | nonhuman; animal cell; mouse; animal; mice; tumor cells, cultured; mice, inbred strains; gene expression regulation; molecular cloning; cloning, molecular; blotting, western; antibodies, monoclonal; molecular sequence data; genetic stability; base sequence; nucleic acid hybridization; blotting, northern; drug stability; receptor binding; oligonucleotides, antisense; delta opiate receptor; receptors, opioid, delta; antisense oligodeoxynucleotide; oligonucleotide probes; antisense oligodeoxynucleotides; male; priority journal; article; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; northern blot; solution hybridization; ng108‐15 cells; western blot; δ‐opioid receptor |
| Journal Title: | Journal of Neurochemistry |
| Volume: | 65 |
| Issue: | 5 |
| ISSN: | 0022-3042 |
| Publisher: | Wiley Blackwell |
| Date Published: | 1995-11-01 |
| Start Page: | 1981 |
| End Page: | 1987 |
| Language: | English |
| DOI: | 10.1046/j.1471-4159.1995.65051981.x |
| PUBMED: | 7595481 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 28 August 2018 -- Source: Scopus |
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