Adaptive supersensitivity in the guinea pig vas deferens is associated with a reduction in the abundance of the alpha 2 subunit isoform of Na+/K(+)-ATPase.
Adaptive supersensitivity has been demonstrated previously in the guinea pig vas deferens after chronic treatment with reserpine, postganglionic denervation, or preganglionic denervation. The magnitude of the change in sensitivity was similar regardless of the method of induction; the underlying mechanism was identified as a partial depolarization secondary to reduced activity of the Na+/K+ pump. Experiments were conducted to quantitatively determine whether the identified losses in Na+/K(+)-ATPase activity and [3H]ouabain binding were due to reductions in the levels of specific protein subunits of the sodium pump. Electrophoretic separation and quantification of the abundance of alpha subunit isoforms were accomplished using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and slot blot analysis. Supersensitivity was induced in the guinea pig vas deferens through pretreatment with reserpine (1.0 mg/kg/day x 5 days). The abundance of the alpha 2 subunit isoform was reduced by 41% in tissue homogenates obtained from animals treated with reserpine, compared with untreated controls. In contrast, there was no significant alteration in the alpha 1 subunit isoform (a protein similar in size to that previously identified in vascular smooth muscle as a "truncated" form of the protein). These data suggest that the adaptation of the guinea pig vas deferens after a chronic reduction in net stimulus is mediated through a change in a specific cellular protein. This evidence supports the assignment of the alpha 2 subunit isoform as the specific protein responsible for the development of nonspecific adaptive supersensitivity in the guinea pig vas deferens.
Hershman
K M
KM
Department of Pharmacology and Toxicology, West Virginia University Health Sciences Center, Morgantown 26506-9223.
Taylor
D A
DA
Fleming
W W
WW
D000818:Animals; D000911:Antibodies, Monoclonal; D001693:Biological Transport, Active; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D015151:Immunoblotting; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D012110:Reserpine / Q000494:pharmacology*; D012964:Sodium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*; D014649:Vas Deferens / Q000187:drug effects* / Q000201:enzymology
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D012110:Reserpine; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase