Cellular and subcellular immunolocalization of alpha1-fetoprotein and albumin in rat liver. Reevaluation of various experimental conditions.

Bélanger, Luc; Beaumont, Carole; Valet, Jean-Paul; Briggs, Robert C.; Chiu, Jen‐Fu

doi:10.1177/26.11.82574

Cited by 63 publications

(21 citation statements)

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“…This pattern is somewhat reminiscent of early reports of immunostaining for albumin (29,30), which also showed scattered positive cells and initially suggested the possibility of functional diversity in production of albumin by hepatocytes, with overexpression in some cells allowing other hepatocytes to more actively produce other needed proteins. However, subsequent studies did not confirm this hypothesis (31).…”

Section: Discussionsupporting

confidence: 74%

Hepatocellular Carcinomas Show Abnormal Expression of Fibronectin Protein

et al. 2002

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Fibronectin plays an important role in cell-to-cell adhesion, cell migration, and cell signaling. In the liver, fibronectin expression has been studied primarily as a component of the extracellular matrix, but little information is available on the expression of fibronectin protein in the neoplastic cells of hepatocellular carcinomas (HCCs). Twenty-four surgically resected HCCs were immunostained with fibronectin. Tumor and normal liver tissues were concurrently analyzed in all cases, and expression in the tumor was evaluated in comparison to the nonneoplastic liver. The average age at resection was 54 ؎ 18 years for the 18 men and 6 women. Twenty-one of the cases were classic HCCs including 6 cases that were well differentiated, 12 cases moderately differentiated, and 3 cases poorly differentiated. The remaining 3 cases were moderately differentiated fibrolamellar carcinomas. In the normal liver, fibronectin labeled the sinusoids and weakly to moderately stained the cytoplasm of hepatocytes. In HCCs, 15/24 showed overexpression of fibronectin in the cytoplasm, 8/24 showed no change from the nonneoplastic liver, and one case showed decreased cytoplasmic staining. In addition, an abnormal membranous staining pattern was noted in 16/24 HCCs. In contrast to the HCCs, none of the three fibrolamellar carcinomas showed increased cytoplasmic or membranous staining. Excluding fibrolamellar carcinoma, increased cytoplasmic staining and/or an abnormal membranous staining was noted in 19/21 (90%) of HCCs. Fibronectin shows abnormal cytoplasmic and/or membranous staining in the majority of HCCs. The implications of fibronectin overexpression are uncertain but may reflect a critical step in tumor genesis.

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Section: Discussionsupporting

confidence: 74%

Hepatocellular Carcinomas Show Abnormal Expression of Fibronectin Protein

et al. 2002

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“…The available evidence indicates that albumin synthesis is restricted to the mid S to late G2 phase of the hepatoma cell cycle (Tsukada & Hirai, 1975) while AFP production, at least in some rat )O hepatoma cells and in newborn rat hepatocytes, is a G1 and/or Go event (Guillouzo et al, 1978;, Tsukada & Hirai, 1975 Scott & Florine, 1982) strongly implicate growth arrest at a specific topographic stage in the G1 phase of the cell cycle to be involved in the expression of differentiated cell functions.…”

Section: Discussionmentioning

confidence: 99%

Secretion of albumin and alpha-foetoprotein by dimethylsulphoxide-stimulated hepatocellular carcinoma cells

Higgins¹,

Darzynkiewicz²,

Melamed³

1983

Br J Cancer

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“…Slides were dipped in Kodak NTB-3 nuclear track emulsion melted at 45°C and diluted 1:1 with 600 mM ammonium acetate (26) and were then dried and stored in a light-proof box at 4°C for [1][2][3] The presence of collagen mRNA in the same preparation of hepatocytes was evaluated by in situ hybridization using a chicken pro-a2(I) collagen DNA probe. As shown in Fig.…”

Section: Preparationmentioning

confidence: 99%

“…One use for this in situ hybridization method was determination of the number and distribution of albumin-synthesizing hepatocytes in the liver, which has remained controversial despite immunohistochemical studies (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12). One concern has been the possible back diffusion of serum albumin into hepatocytes during tissue processing and fixation.…”

Section: Preparationmentioning

confidence: 99%

“…Immunohistochemical localization of albumin in liver sections or in isolated hepatocytes has yielded conflicting results regarding the percentage of hepatocytes involved in the synthesis of this protein. Many immunohistochemical studies (1)(2)(3)(4)(5) as well as some employing a hemolytic plaque test (6)(7)(8) suggested that albumin is produced by only a fraction of normal adult hepatocytes. However, recent reports of experiments using the same techniques have claimed that all normal hepatocytes can produce albumin at one given time (9)(10)(11)(12).…”

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confidence: 99%

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Use of in situ hybridization to identify collagen and albumin mRNAs in isolated mouse hepatocytes.

Saber¹,

Zern²,

Shafritz³

1983

Proc. Natl. Acad. Sci. U.S.A.

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We present a simple and improved method for in situ localization of albumin and collagen mRNAs in isolated mouse hepatocytes. The cells were isolated by collagenase perfusion, mincing, and differential centrifugation. Nick-translated 3H-labeled mouse albumin cDNA (pmalb-2) and chicken pro-a2(I) collagen cDNA (pCg45) probes were then hybridized with the cells in silane-treated microcentrifuge tubes. given time (9-12). Some studies suggest that differences in perfusion techniques may strongly influence the results (13). This is particularly relevant for albumin, the most abundant serum protein, because albumin from the bloodstream can readily crosscontaminate cells in liver tissue.Immunohistochemical methods are limited in that they do not provide information regarding the functional state of the protein synthesis machinery or the level of specific mRNA for a given protein in the cell. This information is important in evaluating the synthesis of proteins present at both high and low levels in cells. In contrast to albumin, collagen is a protein of relatively low abundance in the liver and is capable of being induced under certain pathophysiologic conditions. However, controversy exists as to whether collagen is actually synthesized by hepatocytes (14-20). To investigate this question and to evaluate the synthetic potential of hepatocytes for both albumin and collagen, we have employed in situ hybridization to detect the presence or absence of the specific mRNAs for these proteins. The present report describes a simplified method for in situ hybridization that permits accurate and sensitive localization of albumin and collagen mRNAs in hepatocytes isolated from mouse liver. (24), prepared by the same procedures, was used as a control. MATERIALS AND METHODS PreparationPreparation of Hepatocytes. Hepatocytes were isolated from CF1 mice (Charles River Breeding Laboratories). Livers were perfused in situ at 37°C by the technique of Elliott et al. (25) in a medium consisting of 137 mM NaCl, 5.3 mM KCI, 0.8 mM MgSO4, 0.4 mM Na2HPO4, 0.4 mM KH2PO4, 5.5 mM glucose, and 5 mM Hepes, pH 7.4, under continuous oxygenation while the pH was maintained constant. The liver was perfused in a noncirculating system for 2 min at a flow rate of 5 ml/min. The system was then set to recirculate at 5 ml/min for 5 min with the perfusate supplemented with 0.05% collagenase (Sigma type I) and 2.5 mM CaCl2. The liver was then removed, minced in the collagenase-containing buffer, and incubated for 5-10 min in a shaking water bath at 37°C. The suspension was passed through an 80-,tm nylon mesh and the cells were washed three times in the perfusate buffer without collagenase. The cells were then washed by a sedimentation-resuspension procedure utilizing a clinical centrifuge at 800 X g for 30 sec. The supernatant (containing Kupffer cells and damaged hepatocytes) was removed by aspiration. The pellet was suspended in the same buffer and passed through a 30-,Lm nylon mesh. Viability of isolated cells was determined by trypan blue dye e...

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Cellular and subcellular immunolocalization of alpha1-fetoprotein and albumin in rat liver. Reevaluation of various experimental conditions.

Cited by 63 publications

References 0 publications

Hepatocellular Carcinomas Show Abnormal Expression of Fibronectin Protein

Hepatocellular Carcinomas Show Abnormal Expression of Fibronectin Protein

Secretion of albumin and alpha-foetoprotein by dimethylsulphoxide-stimulated hepatocellular carcinoma cells

Use of in situ hybridization to identify collagen and albumin mRNAs in isolated mouse hepatocytes.

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