Deleterious actions of chronic ethanol treatment on the glycosylation of rat brain clusterin.

Clusterin is a N-glycosylated sialoglycoprotein present in rat brain cells. Clusterin, which elicits aggregation in a wide variety of cells, has been suggested to play an important role in synaptic remodeling through its cell adhesion property or lipid transport capacity in the brain. Sialic acid residues in clusterin may be responsible for its structural conformation, stability and functional ability. Maturation of clusterin is governed by the relative actions of sialyltransferases and sialidases that are present in brain microsomes, golgi bodies, cytosol and plasma membranes. We have earlier reported that chronic ethanol treatment in rats has a damaging effect on the hepatic glycosylation machinery. Others have reported increased hydrolysis of brain sialoconjugates in rats following chronic ethanol administration. Specificity of the effects of chronic ethanol treatment in the brain in relation to the glycosylation process, is still obscure. Therefore, in this investigation, we have studied the specific effects of chronic ethanol treatment on the glycosylation of rat brain clusterin and the causes that may lead to any possible defects in the glycosylation process. We have determined the effects of chronic ethanol treatment on (i) the incorporation of labeled leucine and N-acetylmannosamine into immunoprecipitable clusterin in whole brain homogenate, microsomes, golgi, cytosol, plasma membrane and synaptosomes, (ii) enzymatic activities of sialyltransferases in golgi and synaptosomes, and sialidase in brain cytosol and plasma membranes, and (iii) de novo synthetic rate of rat brain cytosolic sialidase. Our results showed that chronic ethanol treatment in rats resulted in (1) a decreased sialation index of brain clusterin by 47. 2% (p<0.001), 56.7% (p<0.05), 51.7% (p<0.05), 64.8% (p<0.001), and 54.5% (p<0.05), respectively, in whole brain homogenate, golgi, cytosol, plasma membranes, and synaptosomes; (2) a 46.1% (p<0.05) and 12.5% (p<0.05) decreased activities of brain sialyltransferases, respectively, in the golgi and the synaptosomal fractions; (3) a 70. 1% (p<0.05) and 42.6% (p<0.05) increased activities of sialidases, respectively, in the cytosol and plasma membrane fractions; and (4) a 22.2%-64.3% (p<0.001) increased incorporation of labeled leucine into brain cytosolic sialidase. Our findings have clearly established that long-term ethanol treatment in rats leads to a marked impairment in the glycosylation of rat brain clusterin as a result of altered activities of brain sialation and desialation enzymes. In particular, the specific increase noted in brain sialidase activity was due to concomitant increases in its synthetic rate. These defects in the glycosylation of brain clusterin may lead to changes in the molecular conformation of clusterin, and thus, may result in its structural instability and/or functional impairment.