ULTRACYTOCHEMICAL ALTERATIONS IN RAT LIVER MICROSOMES TREATED WITH PHOSPHOLIPASES

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Adult female rats were sacrificed after an 18-hr fast or 6 hr after an injection of ethionine at the 12th-hr of fasting, and both blocks of the hepatic tissue and pellets of the liver microsome fraction were prefixed in cold Karnovsky's fixative for 2-3 hr and washed in cold cacodylate buffer with sucrose overnight. Floating thick sections were treated with enzymic digestive methods using purified snake venom phospholipase A_2 or bacillus phospholipase C. A pale lipid particles with a few reaction products developed in many ER of the samples from only ethionine-injured livers, where cytoplasmic lipid inclusions without the reaction products increased. Independent of the development of fatty livers, minute electron-dense deposits with many reaction products were observed sparsely only on phospholipase-treated samples. They appeared as a granularly dark globule protruding from some biomembranes of phospholipase A_2-treated materials or as an amorphously dark, comma-shaped feature associated with some biomembranes of phospholipase C-treated materials. Therefore, the newly appearing deposits in the phospholipase digestive methods result from the digestion of some intracytoplasmic membrane phospholipids.