[Functional coupling between neuron and glia in neuronal sphingolipid biosynthesis].
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Ceramide lactoside [1-O-(galactosido-4-beta-glucosido)-2-N-acyl-sphingosine] was hydrolysed to ceramide glucoside and galactose by beta-galactosidase of rat brain. The reaction was not reversible, required cholate or taurocholate, had optimum pH5.0 and K(m) 2.2x10(-5)m. It was inhibited by gamma-galactonolactone and galactose as well as by ceramide, sphingosine and fatty acid. Ceramide lactoside could be degraded to ceramide, galactose and glucose by mixtures of rat-brain beta-galactosidase and ox-brain beta-glucosidase.
Ceramide glucoside (1-O-glucosido-2-N-acyl-sphingosine) was hydrolysed to ceramide (N-acyl-sphingosine) and glucose by beta-glucosidase from ox brain. The reaction was stimulated by the non-ionic detergent, Triton X-100, or by the anionic detergents, cholate or taurocholate. It was not reversible, had optimum pH5.0 (with acetate buffer) or 5.6 (with pyridine buffer), had K(m) 1.8x10(-4)m and was inhibited by delta-gluconolactone and sphingosine, but not by ceramide or palmitic acid.
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We studied the synthesis and hydrolysis of sphingomyelin by homogenates of aortic intima plus inner media from normal squirrel monkeys and from monkeys with nutritionally-induced atherosclerosis (6-10 mo on a semi-purified diet containing butter and cholesterol). The concentrations of sphingomyelin in the aortas and plasmas of the atherosclerotic monkeys were higher than those for the normal monkeys. Palmitoyl-1-(14)C coenzyme A was actively utilized for the synthesis of ceramide (N-palmitoyl sphingosine). The addition of sphingosylphosphorylcholine increased the utilization of palmitoyl CoA in sphingomyelin synthesis, and the addition of psychosine (sphingosyl galactoside) increased the incorporation of palmitate into cerebrosides. Rates of sphingomyelin and ceramide synthesis were significantly higher in the atherosclerotic than in the control aortas. Hydrolysis of labeled sphingomyelin to ceramide was also increased in homogenates of the atherosclerotic aortas. Labeled sphingomyelin was taken up from plasma by everted carotid arteries, and this process was also enhanced by atherosclerosis. Increased rates of synthesis and of uptake from plasma of sphingomyelin may account for the increased concentrations of sphingomyelin in the atherosclerotic arteries, even though the ability to degrade sphingomyelin is also enhanced in the atherosclerotic aorta.
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