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Identification of Endomorphin‐1 and Endomorphin‐2 Binding Sites in Human μ‐Opioid Receptor by Antisense Oligonucleotide Strategy
Author(s) -
Fichna Jakub,
Gach Katarzyna,
Perlikowska Renata,
Poels Jeroen,
Vanden Broeck Jozef,
Szemraj Janusz,
Janecka Anna
Publication year - 2008
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/j.1747-0285.2008.00725.x
Subject(s) - exon , receptor , oligonucleotide , aequorin , opioid receptor , microbiology and biotechnology , chemistry , oligopeptide , calcium in biology , intracellular , biology , biochemistry , peptide , opioid , gene
The effects of phosphorothioate antisense oligodeoxynucleotides against exons‐1, ‐2, ‐3 and ‐4 of the human μ‐opioid receptor were studied in the CHO‐μ‐opioid receptor cells using aequorin luminescence‐based calcium assay. All four antisense oligodeoxynucleotides significantly decreased the level of μ‐opioid receptor mRNA in comparison with the non‐treated cells, used as control. However, no statistically significant differences between antisense oligodeoxynucleotides were observed. antisense oligodeoxynucleotides against exon‐2 attenuated endomorphin‐1‐induced intracellular calcium response in a concentration‐dependent manner. antisense oligodeoxynucleotides against exons‐1, ‐2, ‐3 and ‐4 inhibited endomorphin‐2‐induced intracellular calcium response in a concentration‐dependent manner and the effect of antisense oligodeoxynucleotides against exons‐3 and ‐4 was most pronounced. The mismatch oligodeoxynucleotides against respective exons failed to exert any effect. The selective actions of antisense probes directed against different exons of the human μ‐opioid receptor gene, that resulted, at the protein level, in attenuation of calcium responses induced by endomorphin‐1 and endomorphin‐2, suggest that the binding sites for endomorphins are structurally and functionally different. The presence of functionally distinct binding sites might play a crucial role in the modulation of pain and may be important clinically.