Continuity of intracellular channels with extracellular space in adipose tissue and liver: Demonstrated with tannic acid and lanthanum

Abstract
Tannic acid was used to demonstrate continuity of intracellular channels with extracellular space in white adipose tissue of adult rats, brown adipose tissue of suckling rats, and liver of diabetic rats. Electron-opaque material resulting from treatment of glutaraldehyde-fixed tissue with tannic acid was found in extracellular space, invaginations of cell surfaces, vesicles, and intracellular channels. Electron-opaque material was present in channels that surrounded lipid droplets in both white and brown adipocytes and in hepatocytes. The small distance between the lumen of marked channels and lipid droplets in adipocytes indicates that a monolayered structure, perhaps a leaflet of membrane lining the channel, separates the lipid droplet from the lumen of the channel, suggesting that the lipid droplet may be located between leaflets of the membrane lining the channel. Similar findings were obtained in brown adipose tissue using lanthanum instead of tannic acid to mark intracellular channels continuous with extracellular space. Since endoplasmic reticulum is the primary site of triacylglycerol synthesis in adipocytes, marked channels near lipid droplets may be elements of endoplasmic reticulum. Some of the channels marked with tannic acid in hepatocytes contained lipoprotein particles, whereas others were located, in relation to mitochondria and lipid droplets, in the same sites as endoplasmic reticulum in untreated tissue. This indicates that some of the channels marked with tannic acid in hepatocytes are endoplasmic reticulum. Presence of electron-opaque material in intracellular channels and vesicles, but not in cytoplasm, of treated tissue indicates the channels and vesicles were open to extracellular space during treatment with tannic acid or lanthanum and, furthermore, that their membranes were continuous with plasma membrane.