Nicotinic receptor subunit alpha5 modifies assembly, up-regulation, and response to pro-inflammatory cytokines.
In the mammalian brain high affinity nicotine-binding sites are composed of at least the alpha4 and beta2 subunits. Additional nicotinic acetylcholine receptor subunits that are often co-expressed with alpha4+beta2 include alpha5. The introduction of alpha5 into 293 cells expressing alpha4+beta2 strongly favors assembly of alpha4+alpha5+beta2 receptors, increases constitutive ligand binding density as measured using [(3)H]epibatidine, but reduces the magnitude of up-regulation in response to chronic nicotine. In contrast, when beta4 is substituted for beta2, alpha5 interferes with the assembly of these receptors, demonstrating an important role for the beta subunit in this process. When cells co-express alpha4+alpha5+beta2+beta4, over 50% of the subunit associations include all four subunits, but they fail to be detected using [(3)H]epibatidine binding. However, complexes of alpha4+alpha5+beta2 do preferentially emerge from these subunit mixtures, and these mixtures bind ligand. In previous studies of alpha4+beta2+beta4 co-expression by 293 cells, the inflammatory cytokines IL-1beta and TNFalpha influenced the outcome of receptor assembly (Gahring, L. C., Days, E. L., Kaasch, T., González de Mendoza, M., Owen, L., Persiyanov, K., and Rogers, S. W. (2005) J. Neuroimmunol. 166, 88-101). When alpha5 is included in this subunit mixture, and cells are exposed to either inflammatory cytokine, subunit association is no longer altered. These findings suggest that alpha5 is an influential modulator of alpha4+beta2 nicotinic acetylcholine receptor assembly and stabilizes their expression in response to fluctuations in external conditions.
Gahring
Lorise C
LC
Salt Lake City Veterans Affairs-Geriatrics Research, Education and Clinical Center, University of Utah, Salt Lake City, Utah 84132, USA.
Rogers
Scott W
SW
D000818:Animals; D016207:Cytokines / Q000494:pharmacology*; D007155:Immunologic Factors / Q000494:pharmacology; D053583:Interleukin-1beta / Q000494:pharmacology; D055550:Protein Stability; D021122:Protein Subunits / Q000378:metabolism; D051381:Rats; D011978:Receptors, Nicotinic / Q000096:biosynthesis* / Q000378:metabolism / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015854:Up-Regulation
D016207:Cytokines; D007155:Immunologic Factors; D053583:Interleukin-1beta; D021122:Protein Subunits; D011978:Receptors, Nicotinic; D014409:Tumor Necrosis Factor-alpha; C499978:Chrna5 protein, rat