Molecular isoforms of Na+/K(+)-ATPase in the nervous system and kidney of the spontaneously diabetic BB/Wor rat.
Na+/K(+)-ATPase was evaluated in the retina and kidney of the spontaneously diabetic BB/Wor rat after 1 and 4 months of insulin dependency. Retinal synthesis of the Na+/K(+)-ATPase was measured during a 2-h intravitreal pulse of [35S]methionine and analyzed by SDS-PAGE and scintillation counting. Synthesis of the alpha-1 and 'alpha(+)' (includes both alpha-2 and alpha-3) isoforms of the catalytic subunit was increased 123% and 69%, respectively at 4 months. Increases were also suggested at 1 month, but were not significant. The diabetes-dependent peak of synthesis in long-term diabetic rats turned over rapidly and by 3 days after intravitreal labeling, radioactively labeled enzyme was equal in both control and diabetic retinae. The amount of axonally transported, labeled enzyme recovered from endings of the optic nerve in the superior colliculus paralleled retinal labeling. Significant renal hypertrophy (48%) was noted at 4 months, but not at 1 month. The strophanthidin-inhibition constant for diabetes-induced renal enzyme was the same as for control enzyme (approx. 10(-4) M), indicating that diabetic renal hypertrophy does not induce a Na pump isozyme that is more sensitive to cardiotonic steroids. SDS-PAGE of the renal enzyme also failed to indicate more than one isoform of the alpha subunit.
Specht
S C
SC
Department of Pharmacology, University of Puerto Rico, San Juan.
Martin
J
J
Gaud
R E
RE
De Hoyos
J
J
D000818:Animals; D001369:Axons / Q000201:enzymology; D003922:Diabetes Mellitus, Type 1 / Q000188:drug therapy / Q000201:enzymology*; D007328:Insulin / Q000008:administration & dosage; D007527:Isoenzymes / Q000378:metabolism; D007668:Kidney / Q000201:enzymology*; D009441:Neurilemma / Q000201:enzymology; D051381:Rats; D011913:Rats, Inbred BB; D012160:Retina / Q000201:enzymology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry*; D013477:Superior Colliculi / Q000201:enzymology; D013997:Time Factors
D007328:Insulin; D007527:Isoenzymes; D000254:Sodium-Potassium-Exchanging ATPase