title: Evidence against impaired brain microtubule protein polymerization at high glucose concentrations or during diabetes mellitus.
abstract: Previous studies suggest that brain microtubule protein exposed to high glucose levels or isolated from diabetic rats can become glucosylated and that this impairs GTP-induced microtubule polymerization. We set out to extend that investigation to define the mechanistic basis for inhibition of microtubule assembly during diabetes or on incubation at high glucose levels. Rat and bovine brain microtubule protein was purified by cycles of polymerization/depolymerization. When microtubules were incubated for 1 h in either buffer or buffer containing glucose (up to 165 mM), there was no difference in polymerization, a finding contrary to the earlier study. Other rats were injected with vehicle or streptozotocin (90 mg/kg) to induce diabetes as evidenced by serum glucose in excess of 300 mg%, and at 4 weeks, brain microtubule protein was isolated by the polymerization cycling method. Again, there was no difference in the amount or purity of isolated microtubule protein between control or diabetic rats. We also observed no increase in microtubule glucosylation, and GTP-induced polymerization in vitro was indistinguishable for protein derived from brains of normal rats and rats with diabetes as measured by turbidity or electron microscopy. Our results suggest that in vitro incubation with glucose or in vivo elevation of glucose during diabetes fails to impair microtubule polymerization, pointing to other mechanisms for the neuropathy associated with diabetes.
authors.affiliation: Department of Medicine, University of Florida College of Medicine, Gainesville 32610.
authors.forename: E Y
authors.initials: EY
authors.lastname: Eaker
authors.forename: J M
authors.initials: JM
authors.lastname: Angelastro
authors.forename: D L
authors.initials: DL
authors.lastname: Purich
authors.forename: C A
authors.initials: CA
authors.lastname: Sninsky
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D002417:Cattle; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D008868:Microtubule Proteins / Q000378:metabolism*; D008870:Microtubules / Q000648:ultrastructure; D009994:Osmolar Concentration; D011108:Polymers / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D008868:Microtubule Proteins; D011108:Polymers; D005947:Glucose