Differential effects of hypoxia on ligand binding properties of nicotinic and muscarinic acetylcholine receptors on cultured bovine adrenal chromaffin cells.
Hypoxia is known to disturb neuronal signal transmission at the synapse. Presynaptically, hypoxia is reported to suppress the release of neurotransmitters, but its postsynaptic effects, especially on the function of neurotransmitter receptors, have not yet been elucidated. To clarify the postsynaptic effects, we used cultured bovine adrenal chromaffin cells as a model of postsynaptic neurons and examined specific binding of L-[3H]nicotine (an agonist for nicotinic acetylcholine receptors: nAChRs) and 22Na+ flux under control and hypoxic conditions. Experiments were performed in media preequilibrated with a gas mixture of either 21% O2/79% N2 (control) or 100% N2 (hypoxia). Scatchard analysis of the specific binding to the cells revealed that the KD under hypoxic conditions was twice as large as that under control conditions, whereas the Bmax was unchanged. When the specific [3H]nicotine binding was kinetically analyzed, the association constant (k1) but not the dissociation constant (k-1) was decreased to 40% of the control value by hypoxia. When the binding assay was performed using the membrane fraction, these changes were not observed. Nicotine-evoked 22Na+ flux into the cells was suppressed by hypoxia. In contrast, specific [3H]quinuclidinyl benzilate binding to the intact cells was unaffected by hypoxia. These results demonstrate that hypoxia specifically suppresses the function of nAChRs (and hence, neuronal signal transmission through nAChRs), primarily by acting intracellularly.
Lee
K
K
Department of Physiology, Gifu University School of Medicine, Japan.
Ito
A
A
Koshimura
K
K
Ohue
T
T
Takagi
Y
Y
Miwa
S
S
D000311:Adrenal Glands / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002838:Chromaffin System / Q000166:cytology / Q000378:metabolism*; D000860:Hypoxia / Q000378:metabolism*; D008024:Ligands; D009538:Nicotine / Q000378:metabolism; D011813:Quinuclidinyl Benzilate / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism
D008024:Ligands; D011976:Receptors, Muscarinic; D011978:Receptors, Nicotinic; D011813:Quinuclidinyl Benzilate; D009538:Nicotine; D012964:Sodium