Go mediates the coupling of the mu opioid receptor to adenylyl cyclase in cloned neural cells and brain.

Abstract

In membranes from SH-SY5Y human neuroblastoma cells differentiated with retinoic acid, the mu-selective agonist Tyr-D-Ala-Gly-N-Me-Phe-Gly-ol (DAMGO) inhibited cAMP formation with an IC50 of 26 nM. Two separate antibodies raised against distinct regions of the Go alpha sequence attenuated the effect of DAMGO by 50-60%, whereas antibodies to Gi alpha 1,2 or Gi alpha 3 reduced the mu-opioid signal insignificantly or to a lesser extent. In contrast, inhibition of adenylyl cyclase by the delta-opioid agonist Tyr-D-Pen-Gly-Phe-D-Pen-OH (DPDPE; Pen = penicillamine) was very sensitive to the Gi alpha 1,2 antibody. In membranes from rat brain striatum, coupling of the mu opioid receptor to adenylyl cyclase was also maximally blocked by antibodies to Go alpha. After long-term treatment of the cells with DAMGO, the content of Go alpha was reduced by 26%, whereas the levels of Gi alpha 1,2, Gi alpha 3, and Gs alpha were unaltered. Addition of Go, purified from bovine brain, to membranes from pertussis toxin-treated SH-SY5Y cells restored the inhibition of adenylyl cyclase by DAMGO to 70% of that in toxin-untreated cells. To comparably restore the effect of DPDPE, much higher concentrations of Go were required. By demonstrating mediation of cAMP-dependent signal transduction by Go, these results describe (i) an additional role for this G protein present at a high concentration in brain, (ii) preferential, although not exclusive, interaction of mu and delta opioid receptors with different G protein subtypes in coupling to adenylyl cyclase, and (iii) reduced levels of Go following chronic opioid treatment of SH-SY5Y cells with mu opioids.

• Authors: B D Carter, F Medzihradsky
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 90 Issue 9 Pg. 4062-6 (May 01 1993) ISSN: 0027-8424 [Print] United States
• PMID: 8097884 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Adenylate Cyclase Toxin
• Analgesics
• Antibodies
• Enkephalins
• Membrane Proteins
• Receptors, Opioid, mu
• Virulence Factors, Bordetella
• Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
• phenylalanyl-cyclo(cysteinyltyrosyl-tryptophyl-ornithyl-threonyl-penicillamine)threoninamide
• Somatostatin
• Enkephalin, D-Penicillamine (2,5)-
• Cyclic AMP
• Pertussis Toxin
• GTP-Binding Proteins
• Adenylyl Cyclases

Topics

• Adenylate Cyclase Toxin
• Adenylyl Cyclases (metabolism)
• Analgesics (pharmacology)
• Animals
• Antibodies (pharmacology)
• Cell Membrane (drug effects, enzymology, metabolism)
• Clone Cells
• Corpus Striatum (metabolism)
• Cyclic AMP (metabolism)
• Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
• Enkephalin, D-Penicillamine (2,5)-
• Enkephalins (pharmacology)
• GTP-Binding Proteins (immunology, metabolism)
• Humans
• Kinetics
• Membrane Proteins (isolation & purification, metabolism)
• Neuroblastoma (metabolism)
• Pertussis Toxin
• Rats
• Receptors, Opioid, mu (metabolism)
• Somatostatin (analogs & derivatives, pharmacology)
• Tumor Cells, Cultured
• Virulence Factors, Bordetella (pharmacology)