In Vitro Effects of Ligand Bias on Primate Mu Opioid Receptor Downstream Signaling
Abstract
:1. Introduction
2. Results
2.1. Downstream Signaling Pathway Responses to MOR Activation
2.2. Effects of MOR Activation on NF-ĸB, MAPK/JNK, and cAMP Pathways Across Ligands
2.3. Ligand Bias and Species Differences Exist for MORs
3. Discussion
4. Materials and Methods
4.1. Plasmids
4.2. Cell Lines
4.3. GPCR Signaling Reporter Array
4.4. Drugs
4.5. Concentration Response Determinations
4.6. Data Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
MOR | µ opioid receptor |
NHP | nonhuman primate |
GPCR | G-protein coupled receptor |
cAMP | cyclic adenosine 5′-monophosphate |
NF-ĸB | nuclear factor kappa-light-chain-enhancer of activated B cells |
MAPK | mitogen-activated protein kinase |
JNK | Jun N-terminal kinase |
ERK | extracellular-signal-regulated kinase |
ATF | activating transcription factor |
DAMGO | [D-Ala2-MePhe4-Gly-ol]-enkephalin |
PMA | phorbol 12-myristate 13-acetate |
TNF- α | tumor necrosis factor α |
GFP | green fluorescent protein |
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Ligand | Human | Rhesus Macaque | ||
---|---|---|---|---|
LogEC50 | Emax | LogEC50 | Emax | |
cAMP | ||||
DAMGO | −8.76 ± 0.07 | 100.0 ± 4.09 | −9.04 ± 0.03 | 100.2 ± 6.06 |
morphine | −8.33 ± 0.13 | 100.0 ± 5.35 | −8.57 ± 0.08 | 100.9 ± 7.18 |
met-enkephalin | −8.74 ± 0.07 | 100.4 ± 14.99 | −9.42 ± 0.07 | 100.0 ± 5.09 |
endomorphin-1 | −8.30 ± 0.10 | 100.4 ± 12.14 | −9.14 ± 0.05 | 99.6 ± 6.72 |
β-endorphin | −7.70 ± 0.05 | 100.1 ± 3.70 | −7.50 ± 0.03 | 104.2 ± 1.89 |
TRV130 | −8.85 ± 0.12 | 100.1 ± 6.90 | −9.25 ± 0.07 | 100.0 ± 5.34 |
NF-ĸB | ||||
DAMGO | −9.18 ± 0.07 | 100.0 ± 5.22 | −9.23 ± 0.09 | 100.0 ± 7.72 |
morphine | −8.41 ± 0.05 | 83.2 ± 4.73 | −8.72 ± 0.26 | 93.5 ± 11.34 |
met-enkephalin | −8.08 ± 0.04 | 104.8 ± 5.06 | −8.67 ± 0.16 | 93.7 ± 4.68 |
endomorphin-1 | −8.46 ± 0.15 | 104.2 ± 6.43 | −7.54 ± 0.04 | 104.2 ± 5.21 |
β-endorphin | −7.23 ± 0.07 | 103.0 ± 12.87 | −7.33 ± 0.07 | 103.0 ± 10.71 |
TRV130 | −6.34 ± 0.10 | 159.6 ± 12.61 | −6.61 ± 0.13 | 132.7 ± 11.57 |
MAPK/JNK | ||||
DAMGO | −7.40 ± 0.08 | 101.8 ± 8.47 | −7.88 ± 0.10 | 112.9 ± 10.59 |
morphine | −8.18 ± 0.07 | 90.8 ± 4.59 | −7.86 ± 0.12 | 91.4 ± 7.10 |
met-enkephalin | −7.93 ± 0.08 | 72.0 ± 4.58 | −7.53 ± 0.09 | 73.5 ± 5.33 |
endomorphin-1 | −8.08 ± 0.06 | 90.9 ± 3.84 | −8.99 ± 0.09 | 92.5 ± 3.49 |
β-endorphin | −7.20 ± 0.07 | 102.0 ± 8.43 | −7.71 ± 0.12 | 88.5 ± 7.70 |
TRV130 | −6.99 ± 0.07 | 101.3 ± 10.05 | −6.93 ± 0.08 | 123.2 ± 8.65 |
Ligand | Human | Rhesus Macaque | ||||
---|---|---|---|---|---|---|
cAMP | ||||||
DAMGO | 10.76 ± 0.18 | 0.00 ± 0.25 | 0.00 ± 0.36 | 11.04 ± 0.12 | 0.00 ± 0.17 | 0.00 ± 0.24 |
morphine | 10.33 ± 0.31 | −0.43 ± 0.36 | 0.00 ± 0.51 | 10.59 ± 0.21 | −0.45 ± 0.25 | 0.00 ± 0.35 |
met-enkephalin | 10.72 ± 0.26 | −0.04 ± 0.32 | 0.00 ± 0.45 | 11.41 ± 0.18 | 0.37 ± 0.21 | 0.00 ± 0.30 |
endomorphin-1 | 10.30 ± 0.30 | −0.46 ± 0.35 | 0.00 ± 0.49 | 11.09 ± 0.15 | 0.05 ± 0.19 | 0.00 ± 0.27 |
β-endorphin | 9.70 ± 0.13 | −1.06 ± 0.22 | 0.00 ± 0.32 | 9.50 ± 0.07 | −1.54 ± 0.14 | 0.00 ± 0.20 |
TRV130 | 10.90 ± 0.30 | 0.14 ± 0.35 | 0.00 ± 0.49 | 11.23 ± 0.19 | 0.18 ± 0.22 | 0.00 ± 0.32 |
NF-ĸB | ||||||
DAMGO | 11.18 ± 0.19 | 0.00±0.26 | 0.00 ± 0.37 | 11.23 ± 0.24 | 0.00 ± 0.34 | 0.00 ± 0.38 |
morphine | 10.33 ± 0.15 | −0.85 ± 0.24 | −0.42 ± 0.43 | 10.69 ± 0.62 | −0.54 ± 0.66 | −0.09 ± 0.71 |
met-enkephalin | 10.10 ± 0.11 | −1.08 ± 0.22* | −1.04 ± 0.39 | 10.64 ± 0.36 | −0.59 ± 0.43 | −0.95 ± 0.48 |
endomorphin-1 | 10.48 ± 0.36 | −0.70 ± 0.41 | −0.24 ± 0.53 | 9.55 ± 0.11 | −1.68 ± 0.27 | −1.73 ± 0.33 |
β-endorphin | 9.24 ± 0.25 | −1.94 ± 0.32 | −0.88 ± 0.39 | 9.34 ± 0.23 | −1.88 ± 0.34 | −0.34 ± 0.37 |
TRV130 | 8.54 ± 0.26 | −2.64 ± 0.32 | −2.78 ± 0.48 | 8.74 ± 0.34 | −2.49 ± 0.41 | −2.67 ± 0.47 |
MAPK/JNK | ||||||
DAMGO | 9.41 ± 0.23 | 0.00 ± 0.32 | 0.00 ± 0.41 | 9.93 ± 0.29 | 0.00 ± 0.41 | 0.00 ± 0.44 |
morphine | 10.14 ± 0.19 | 0.74 ± 0.29 | 1.16 ± 0.46 | 9.82 ± 0.31 | −0.11 ± 0.42 | 0.34 ± 0.49 |
met-enkephalin | 9.79 ± 0.22 | 0.39 ± 0.31 | 0.42 ± 0.45 | 9.39 ± 0.25 | −0.54 ± 0.38 | −0.90 ± 0.44 |
endomorphin-1 | 10.04 ± 0.17 | 0.64 ± 0.28 | 1.10 ± 0.45 | 10.96 ± 0.22 | 1.03 ± 0.36 | 0.98 ± 0.41 |
β-endorphin | 9.21 ± 0.20 | −0.19 ± 0.31 | 0.87 ± 0.38 | 9.66 ± 0.32 | −0.27 ± 0.43 | 1.27 ± 0.45 |
TRV130 | 8.99 ± 0.22 | −0.41 ± 0.32 | −0.55 ± 0.47 | 9.02 ± 0.22 | −0.91 ± 0.36 | −1.10 ± 0.42 |
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Zhang, X.; Hutchins, S.D.; Blough, B.E.; Vallender, E.J. In Vitro Effects of Ligand Bias on Primate Mu Opioid Receptor Downstream Signaling. Int. J. Mol. Sci. 2020, 21, 3999. https://doi.org/10.3390/ijms21113999
Zhang X, Hutchins SD, Blough BE, Vallender EJ. In Vitro Effects of Ligand Bias on Primate Mu Opioid Receptor Downstream Signaling. International Journal of Molecular Sciences. 2020; 21(11):3999. https://doi.org/10.3390/ijms21113999
Chicago/Turabian StyleZhang, Xiao, Shaurita D. Hutchins, Bruce E. Blough, and Eric J. Vallender. 2020. "In Vitro Effects of Ligand Bias on Primate Mu Opioid Receptor Downstream Signaling" International Journal of Molecular Sciences 21, no. 11: 3999. https://doi.org/10.3390/ijms21113999