Detection of metabolic changes in hepatocytes by quantitative cytochemistry

James, J.; Frederiks, Wilma M.; Noorden, C. J. F. Van; Tas, Johan

doi:10.1007/bf00482955

Cited by 12 publications

(2 citation statements)

References 73 publications

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“…We applied insulin ex vivo directly into the liver thought the vena porta but could not see a gradient in insulin binding along the periportal-pericentral axis (Supplementary Figure 3 ). There was no indication that nuclear polyploidy differ along this axis which is in agreement with an earlier study (James et al, 1986 ) arguing that the metabolic zonation and the ploidy of liver cells are independent biological features.…”

Section: Discussion and Summarysupporting

confidence: 92%

Hepatocyte Ploidy Is a Diversity Factor for Liver Homeostasis

et al. 2017

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Polyploidy, the existence of cells containing more than one pair of chromosomes, is a well-known feature of mammalian hepatocytes. Polyploid hepatocytes are found either as cells with a single polyploid nucleus or as multinucleated cells with diploid or even polyploid nuclei. In this study, we evaluate the degree of polyploidy in the murine liver by accounting both DNA content and number of nuclei per cell. We demonstrate that mouse hepatocytes with diploid nuclei have distinct metabolic characteristics compared to cells with polyploid nuclei. In addition to strong differential gene expression, comprising metabolic as well as signaling compounds, we found a strongly decreased insulin binding of nuclear polyploid cells. Our observations were associated with nuclear ploidy but not with total ploidy within a cell. We therefore suggest ploidy of the nuclei as an new diversity factor of hepatocytes and hypothesize that hepatocytes with polyploid nuclei may have distinct biological functions than mono-nuclear ones. This diversity is independent from the well-known heterogeneity related to the cells' position along the porto-central liver-axis.

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Section: Discussion and Summarysupporting

confidence: 92%

Hepatocyte Ploidy Is a Diversity Factor for Liver Homeostasis

et al. 2017

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“…Liver fragments, 5 mm3 maximally, were frozen in liquid nitrogen and stored at -80°C. A period of 60min of ischaemia induces necrosis in approximately [10][11][12][13][14][15] per cent of the parenchymal cells of the clamped liver lobes. 6 In the case of ischaemia in vitro, small liver fragments of 5mm in maximum dimension were wrapped in incise drape (Barrier Surgikos, Johnson-Johnson Company, New Brunswick, U.S.A.) and incubated at 37°C for 0, 30, 60, 120, and 240 min.'…”

Section: Methodsmentioning

confidence: 99%

A quantitative histochemical study of 5′nucleotidase activity in rat liver after ischaemia

Frederiks

Marx

Myagkaya

1988

The Journal of Pathology

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The lead salt method of Wachstein and Meisel15 has been applied using incubation media containing polyvinyl alcohol for the localization and quantification of 5'-nucleotidase (E.C.3.1.3.5) activity in cryostat sections from rat liver after ischaemia in vitro and ischaemia in vivo followed by different periods of re-perfusion. 5'-Nucleotidase activity at the bile canaliculi, especially in the pericentral areas, had already decreased after 60 min of ischaemia in vitro, although the total activity as measured densitometrically was not changed. After 120-240 min of ischaemia, a significant decrease of the total 5'-nucleotidase activity was found. At that stage, signs of irreversible cell damage were recognized. Short periods of re-perfusion (1 h) after ischaemia in vivo induced a decreased bile canalicular 5'-nucleotidase activity throughout the entire liver, but a restoration after longer periods of re-perfusion was observed (5, 24, and 48 h). Necrotic areas recognized by a decreased lactate dehydrogenase activity after all periods of re-perfusion showed decreased total 5'-nucleotidase activities. A correlation was observed between the decrease in bile canalicular 5'-nucleotidase activity and the disappearance of microvilli of the bile canaliculi. It is concluded that a decrease in the bile canalicular 5'-nucleotidase activity can be used as a very sensitive marker for ischaemic liver cell damage. Assessment of the irreversibility of the cell injury has to be determined using additional parameters such as a decreased lactate dehydrogenase activity.

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Immunocytochemical determination of ploidy class-dependent bromodeoxyuridine incorporation in rat liver parenchymal cells after partial hepatectomy

et al. 1990

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Immunocytochemistry of bromodeoxyuridine (BrdU) incorporated in DNA was performed on cryostat sections of rat liver and on isolated hepatocytes after partial hepatectomy using a two-step labeling technique. The method enabled the detection of S-phase nuclei in both tissue preparations. Quantification of the number of labeled nuclei in sections showed that the number of nuclei in S-phase increased from 0.3% in control liver to about 36% at 24 h after partial hepatectomy. The detection of BrdU in isolated hepatocytes showed the same labeling index of binuclear diploid, mononuclear tetraploid and binuclear tetraploid cells. A special role for mononuclear diploid cells in proliferation did not seem to occur.

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Detection of metabolic changes in hepatocytes by quantitative cytochemistry

Cited by 12 publications

References 73 publications

Hepatocyte Ploidy Is a Diversity Factor for Liver Homeostasis

Hepatocyte Ploidy Is a Diversity Factor for Liver Homeostasis

A quantitative histochemical study of 5′nucleotidase activity in rat liver after ischaemia

Immunocytochemical determination of ploidy class-dependent bromodeoxyuridine incorporation in rat liver parenchymal cells after partial hepatectomy

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