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Insulin stimulation of (/sup 3/H)-ouabain binding to cerebrovascular (Na/sup +/ + K/sup +/)-ATPase

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5333873
;
Brain microvessels were isolated from rat cerebral cortices. The binding of (/sup 3/H)-ouabain to microvascular (Na/sup +/ + K/sup +/)-ATPase increased with microvessel protein (37-110..mu..g) and was time dependent with maximum binding observed at 15 min at 37/sup 0/C. Non-specific binding, measured in the presence of 50..mu..M ouabain, was less than 2% of total binding. Scatchard analysis of preliminary (/sup 3/H)-ouabain binding data yielded a K/sub D/ of 44nM and a B/sub max/ of 12pmol/mg. Since the high affinity (..cap alpha..+) form of the enzyme is purportedly hormonally regulated, the effect of insulin on (/sup 3/H)-ouabain binding to microvessels was studied. Insulin (0.001-10..mu..M) stimulation of (/sup 3/H)-ouabain binding was dose dependent. To assess whether this was a specific or a peptide-protective effect, assays were performed in the presence of bovine serum albumin (BSA). Addition of BSA (10..mu..M) enhanced the amount of (/sup 3/H)-ouabain bound 4-fold. Further increases in the BSA concentration (20..mu..M) did not increase binding. Addition of 10..mu..M insulin evoked a 20% increase in (/sup 3/H)-ouabain binding above BSA-treated controls. In summary, the data suggest that the (..cap alpha..+) form of the (Na/sup +/ + K/sup +/)-ATPase is present in cerebral endothelium and (/sup 3/H)-ouabain binding is significantly elevated by insulin in a dose dependent manner. Therefore, insulin may regulate microvascular (Na/sup +/ + K/sup +/)-ATPase and thus be a modulator of blood-brain permeability to ions.
Research Organization:
Univ. of Detroit, MI
OSTI ID:
5333873
Report Number(s):
CONF-8606151-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACID ANHYDRASES
AFFINITY
ALBUMINS
ANIMAL TISSUES
ANIMALS
ATP-ASE
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BLOOD VESSELS
BODY
BRAIN
CARBOHYDRATES
CARDIAC GLYCOSIDES
CARDIOTONICS
CARDIOVASCULAR AGENTS
CARDIOVASCULAR SYSTEM
CATTLE
CENTRAL NERVOUS SYSTEM
DOMESTIC ANIMALS
DOSE-RESPONSE RELATIONSHIPS
DRUGS
ENDOTHELIUM
ENZYMES
GLYCOSIDES
HORMONES
HYDROLASES
INSULIN
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANS
OUABAIN
PEPTIDE HORMONES
PERMEABILITY
PHOSPHOHYDROLASES
PROTEINS
REACTION KINETICS
RUMINANTS
STROPHANTHINS
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES