Autoradiographic localization of th transport, and catabolism of low de: liver of normal and estrogen-treated (hepatic receptors/endocytosis/multivesicular body-like structures/lys

The hepatic uptake and catabolism of low density lipoproteins are stimulated severalfold in rats treated with large amounts of 17a-ethinylestradiol. To determine the sites within the liver at which these processes occur, '25I-labeled human low den- sity lipoproteins were injected intravenously into intact control and estradiol-treated rats or added to perfusates of their isolated livers. The livers were fixed by perfusion and processed for light and electron microscopic autoradiography. Distribution of auto- radiographic silver grains was estimated qualitatively in light mi- crographs and quantitatively in electron micrographs. Many more silver grains were seen in livers from estradiol-treated than from control rats, but the processing of labeled low density lipoprotein was indistinguishable. Three minutes after intravenous injection or perfusion of livers, the grains were concentrated over the mi- crovillous surface of parenchymal cells bordering the space of Disse. Many of these grains were within two half-distances from endocytic pits. Only 5-15% of the grains were seen over endothe- lial and Kupffer cells. Silver grains were also observed over vesi- cles beneath the plasma membrane whose size and shape sug- gested that they were derived from fusion of endocytic vesicles. By 15 min, grains were predominantly located in structures like mul- tivesicular bodies in the region of the GERL (Golgi com- plex-endoplasmic reticulum-lysosomes) near the bile canaliculi. These bodies were packed with small vesicle-like structures and a few larger vesicles, the latter possessing a unit membrane. Be- tween 15 and 30 min, when proteolysis of low density lipoproteins is known to begin, the initially clear matrix of the multivesicular body-like structures became dark and the structures frequently had a dense tail-like appendage. At the same time, silver grains be- gan to appear over secondary lysosomes. These and other results indicate that the hepatic uptake of low density lipoproteins that is stimulated in rats given large amounts of estradiol follows a path- way that closely resembles that of the well-defined "LDL recep- tor" in cultured cells. In the liver these lipoproteins appear to be transported in endocytic vesicles; the vesicles fuse to form multi- vesicular body-like structures that acquire lysosomal enzymes and are converted to secondary lysosomes as the lipoproteins are degraded.