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Regulation of delta opioid receptor expression by anti‐CD 3 ‐ε, PMA, and ionomycin in murine splenocytes and T cells
Author(s) -
Li Ming D.,
McAllen Kathy,
Sharp Burt M.
Publication year - 1999
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.65.5.707
Subject(s) - ionomycin , splenocyte , biology , δ opioid receptor , microbiology and biotechnology , cd3 , receptor , opioid receptor , immunology , delta , pharmacology , opioid , immune system , biochemistry , intracellular , cd8 , engineering , aerospace engineering
Previous studies have shown that low levels of delta opioid receptor (DOR) mRNA were detectable by reverse transcription polymerase chain reaction (RT‐PCR) in unstimulated splenocytes from BALB/c female mice. This study demonstrates that DOR transcripts can be detected in freshly obtained splenocytes from CD1 female mice as well. The results of studies using quantitative competitive RT‐PCR showed that DOR transcripts in splenic T cells increased from < 1 copy/cell to 22 and 42 copies/cell, respectively, after stimulation with anti‐CD3‐ε for 24 and 48 h compared to the level in freshly obtained T cells. In the presence of actinomycin D, anti‐CD3‐ε did not affect the rate of degradation of DOR mRNA, suggesting that its stability is not altered by anti‐CD3‐ε. After incubation with phorbol myristate acetate (PMA) and ionomycin, the expression of DOR mRNA in splenocytes and T cells was significantly reduced compared with unstimulated cells in culture. In addition, the inhibitory effect of PMA prevented anti‐CD3‐ε‐stimulated DOR expression. These data suggest that signaling through the T cell receptor complex by anti‐CD3‐ε regulates DOR expression differently than PMA and ionomycin. J. Leukoc. Biol. 65: 707–714; 1999.