Effects of antimicrotubule agents on phospholipid metabolism in rat hepatic subcellular membranes

Abstract

Treatment of animals with antimicrotubule drugs has been shown to cause a perplexing variety of cellular changes which, theoretically, coula be the result of changes in endomembrane biosynthesis, composition or flow. In the current study we have focused on this possibility by identifying antimicrotubule drug-induced changes in the phospholipid metabolism of hepatic subcellular membranes. Young adult rats were pretreated with radiolabeled [ 32P]orthophosphate for 12 hr, and subsequently given saline, colchicine (2.5 mg/kg body wt) or vinblastine (20 mg/kg body wt) for 4 additional hr. Afterwards, the livers were homogenized, and separate microsomal and Golgi membrane fractions were prepared and subjected to phospholipid extraction and identification using two-dimensional thin-layer chromatography. The results show that colchicine and vinblastine given in vivo caused specific, rapid and in some cases, dramatic changes in phospholipid turnover in different membrane fractions of rat liver. The drugs specifically increased labeling of phosphatidylinositol-4-monophosphate and phosphatidylinositol-4,5-biphosphate and decreased the radioactivity associated with phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol in all fractions examined. In contrast, the antimicrotubule drugs produced a differential effect on the labeling pattern of sphingomyelin and lysophosphatidylcholine, i.e. they stimulated labeling of these phospholipids in microsomes, produced no changes in heavy Golgi fractions, and markedly increased their labeling in light Golgi fractions. These data suggest that antimicrotubule drugs restrict the incorporation of certain precursor phospholipids into forming membranes but do not affect the subsequent metabolism of these phospholipids. At the same time, the drugs appear to retard the flow of membranes from one cellular compartment to another.