期刊
CHEMICAL BIOLOGY & DRUG DESIGN
卷 72, 期 6, 页码 507-512出版社
WILEY
DOI: 10.1111/j.1747-0285.2008.00725.x
关键词
aequorin luminescence-based calcium assay; endogenous opioid peptide; MOR agonist; real-time RT PCR
资金
- Medical University of Lodz [502-11-460, 502-11-461, 503-10-99-1]
- Bilateral Scientific Cooperation between Flanders and Poland [BIL05/17]
- Foundation for Polish Science
The effects of phosphorothioate antisense oligodeoxynucleotides against exons-1, -2, -3 and -4 of the human mu-opioid receptor were studied in the CHO-mu-opioid receptor cells using aequorin luminescence-based calcium assay. All four antisense oligodeoxynucleotides significantly decreased the level of mu-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 mu-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.
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