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The localization of alpha-fetoprotein (AFP) and of antigens of liver-cell plasma membrane (Ag I, Ag II and Ag III) was studied in adult, postnatal and regenerating (CCl4 or paracetamol-treated mouse liver). Ag I, Ag II and Ag III were solubilized by Triton X-100 from ghosts of the mouse liver cells. The purification of antigens was performed by gel filtration on Sephadex G-200 or Ultrogel ACA-54 with subsequent treatment of antigenic fractions eluted from the column with 5% HClO4. Ag I and Ag II are common for liver and some other mouse organs, but Ag III is strictly specific for liver. The Ag I is predominantly found in the region of the bile capillaries. Ag II and Ag III are mainly present on membrane adjoining blood sinusoids. The distribution of these three antigens in newborn mouse liver is quite different from that in adult liver and reflects the specific arrangement of hepatocytes at different stages of development - the acinar structure in newborn liver and the plate structure in the adult organ. During regeneration as well as during postnatal development, AFP has been found in areas of marked tissue rearrangement. In both cases, these areas lose the plasma membrane antigens, especially Ag I - the antigen of bile capillary. The reappearance of Ag I on the surface of liver cells coincides with cessation of AFP synthesis and establishment of definitive plate structure. The possible role of liver plate structure in the regulation of AFP synthesis is discussed.
The localization of alpha-fetoprotein (AFP) and of antigens of liver-cell plasma membrane (Ag I, Ag II and Ag III) was studied in adult, postnatal and regenerating (CCl4 or paracetamol-treated mouse liver). Ag I, Ag II and Ag III were solubilized by Triton X-100 from ghosts of the mouse liver cells. The purification of antigens was performed by gel filtration on Sephadex G-200 or Ultrogel ACA-54 with subsequent treatment of antigenic fractions eluted from the column with 5% HClO4. Ag I and Ag II are common for liver and some other mouse organs, but Ag III is strictly specific for liver. The Ag I is predominantly found in the region of the bile capillaries. Ag II and Ag III are mainly present on membrane adjoining blood sinusoids. The distribution of these three antigens in newborn mouse liver is quite different from that in adult liver and reflects the specific arrangement of hepatocytes at different stages of development - the acinar structure in newborn liver and the plate structure in the adult organ. During regeneration as well as during postnatal development, AFP has been found in areas of marked tissue rearrangement. In both cases, these areas lose the plasma membrane antigens, especially Ag I - the antigen of bile capillary. The reappearance of Ag I on the surface of liver cells coincides with cessation of AFP synthesis and establishment of definitive plate structure. The possible role of liver plate structure in the regulation of AFP synthesis is discussed.
The AFP-synthesizing cells were identified by ultrastructural localization of the antigen in regenerating liver of adult mice after CCl4 poisoning. An indirect immunoperoxidase method with rabbit anti-mouse AFP and peroxidase conjugates of anti-rabbit IgG or their Fab' was used. Good preservation of AFP and tissue structure, and sufficient permeability for the conjugates were obtained after 20' prefixation of small liver specimens in 8% formaldehyde -0.05% glutaraldehyde followed by 16 h fixation in 8% formaldehyde. The intracellular localization of AFP observed in the light microscope in most cases corresponded to its synthesis and secretion. It was found in two cell types, both concentrated mainly in the perinecrotic zones and constituting only a small part of the whole cell population. Most of the AFP-producing cells were normal differentiated hepatocytes without any structural signs of damage. A few smaller cells with active AFP synthesis were present in some animals. By their ultrastructure they resembled the oval cells found during chemical hepatocarcinogenesis in rats.
The AgB10 antigen of bile canaliculi of the mouse hepatocyte was identified using monoclonal antibodies. The Mr value of 116000 for AgB10 was measured by immunoblotting. The tissue localization of AgB10 was studied by light and electron microscopy using the immunoperoxidase technique. AgB10 was predominantly present on the microvillus membrane of bile canaliculi, the brush border of intestinal mucosa and apical surfaces of the epithelial cells in some other organs. A small amount of AgB10 was detected on the basolateral domain of the hepatocytes. AgB10 was specific for hepatocytes and was not found in the other cell types of the liver. In primary hepatocyte culture, AgB10 was localized on the surface of cells during the first 24 h, predominantly at the sites of cell-cell and cell-substratum contacts. After 48 h of culture AgB10 gradually disappeared from contacting cell surfaces and became concentrated only in the reconstituted bile canaliculi.
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