Hepatocyte ploidy in normal young rat

Gandillet, Arnaud; Alexandre, Eliane; Holl, Vincent; Royer, Corinne; Bischoff, Pierre; Cinqualbre, J; Wolf, Philippe; Jaeck, Daniel; Richert, Lysiane

doi:10.1016/s1095-6433(02)00374-4

Cited by 37 publications

(33 citation statements)

References 31 publications

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“…Tetraploid (XXYY), or higher ploidy cell populations were not found in wild-type E10.5 and E13.5 embryos. The absence of polyploidy in E10.5 and E13.5 embryos is in agreement with studies showing that naturally occurring tetraploidy and polyploidy are not significantly evident until later in postnatal development (Gandillet et al, 2003;Guidotti et al, 2003;Gupta, 2000). FISH analysis on tissues from wild-type P3 mice revealed at least 96% XY diploid cells, with 1.5% or fewer XXYY tetraploid cells.…”

Section: No Difference In Cell Ploidy Distributions In Wild-type Verssupporting

confidence: 90%

“…Liver perfusion through the portal vein consisted of 3 min with EBSS (Earles Basic Salt Solution) without Ca/Mg, 3 min with EBSS with Ca/Mg, and 10 min with 0.5 mg/mL collagenase D (Roche) in EBSS with Ca/Mg. This method is consistent with standard liver perfusion protocols for isolating rat hepatocytes, where hepatocyte polyploidy was assessed using FISH analysis or FACS analysis for DNA content (Gandillet et al, 2003;Gupta, 2000;Wang et al, 2003). Although Wang et al (2003) performed FISH analysis on hepatocytes cultured in vitro for up to 40 h, our results indicate that FISH analysis performed directly after liver perfusion best represents hepatocyte polyploidy, where in vitro hepatocyte culture for 24 and 40 h results in decreased numbers of 6n and 8n hepatocytes (data not shown).…”

Section: Real Time Quantitative Pcr (Qpcr)mentioning

confidence: 89%

“…Tissues such as liver have been shown to have populations of tetraploid or higher ploidy cells that are thought to arise from endoreduplication (Dudas et al, 2003;Guidotti et al, 2003;Gupta, 2000). Although it has not been determined precisely when polyploidy arises in mouse tissues, it has been postulated that liver cell polyploidy begins in rats around 3 weeks of age (Gandillet et al, 2003;Guidotti et al, 2003;Gupta, 2000). The difference in liver cell polyploidy we observed between wild-type and chimeric mice is likely to reflect natural polyploid variability, as it has been shown that hepatocyte polyploidy (4n) in rats can range between 10-20% (Gandillet et al, 2003;Guidotti et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…These observations are consistent with studies in rats showing that hepatocyte polyploidy is first evident at 3 weeks of age. By 6 weeks, polyploid cells are more prevalent than diploid, and at 10-12 weeks, 20% of hepatocytes are diploid and 75% are polyploid (Gandillet et al, 2003;Guidotti et al, 2003).…”

Section: No Difference In Cell Ploidy Distributions In Wild-type Versmentioning

confidence: 99%

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Embryonic Stem Cells Contribute to Mouse Chimeras in the Absence of Detectable Cell Fusion

Kidder¹,

Oseth²,

Miller³

et al. 2008

Cloning and Stem Cells

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Embryonic stem (ES) cells are capable of differentiating into all embryonic and adult cell types following mouse chimera production. Although injection of diploid ES cells into tetraploid blastocysts suggests that tetraploid cells have a selective disadvantage in the developing embryo, tetraploid hybrid cells, formed by cell fusion between ES cells and somatic cells, have been reported to contribute to mouse chimeras. In addition, other examples of apparent stem cell plasticity have recently been shown to be the result of cell fusion. Here we investigate whether ES cells contribute to mouse chimeras through a cell fusion mechanism. Fluorescence in situ hybridization (FISH) analysis for X and Y chromosomes was performed on dissociated tissues from embryonic, neonatal, and adult wild-type, and chimeric mice to follow the ploidy distributions of cells from various tissues. FISH analysis showed that the ploidy distributions in dissociated tissues, notably the tetraploid cell number, did not differ between chimeric and wild-type tissues. To address the possibility that early cell fusion events are hidden by subsequent reductive divisions or other changes in cell ploidy, we injected Z/EG (lacZ/EGFP) ES cells into ACTB-cre blastocysts. Recombination can only occur as the result of cell fusion, and the recombined allele should persist through any subsequent changes in cell ploidy. We did not detect evidence of fusion in embryonic chimeras either by direct fluorescence microscopy for GFP or by PCR amplification of the recombined Z/EG locus on genomic DNA from ACTB-cre::Z/EG chimeric embryos. Our results argue strongly against cell fusion as a mechanism by which ES cells contribute to chimeras. 231

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Section: No Difference In Cell Ploidy Distributions In Wild-type Verssupporting

confidence: 90%

Section: Real Time Quantitative Pcr (Qpcr)mentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Section: No Difference In Cell Ploidy Distributions In Wild-type Versmentioning

confidence: 99%

See 2 more Smart Citations

Embryonic Stem Cells Contribute to Mouse Chimeras in the Absence of Detectable Cell Fusion

Kidder¹,

Oseth²,

Miller³

et al. 2008

Cloning and Stem Cells

View full text Add to dashboard Cite

show abstract

“…In each experiment, some isolated hepatocytes and nuclei were used to determine their respective degree of ploidy as previously described [38]. Briefly, 10 6 hepatocytes were suspended in 1 ml 70% ethanol and kept overnight at 4°C, whereas other 10 6 hepatocytes were submitted to a procedure that extracted almost pure nuclei (with a degree of purity 1 95%) before being similarly stored overnight at 4°C in 1 ml 70% ethanol and then analyzed.…”

Section: Ploidy Analysis By Flow Cytometrymentioning

confidence: 99%

Hepatocyte Ploidy in Regenerating Livers after Partial Hepatectomy, Drug-Induced Necrosis, and Cirrhosis

Gandillet¹,

Alexandre²,

Royer

et al. 2003

Eur Surg Res

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The hepatocyte ploidy was investigated by flow cytometry in regenerating Sprague-Dawley rat livers following either drug-induced acute necrosis (single sublethal doses of D-galactosamine or thioacetamide) or drug-induced chronic cirrhosis (repeated thioacetamide injections for 10–18 weeks) and in regenerating livers following 70% partial hepatectomy and was compared with that of normal hepatocytes. Twenty-four hours after partial hepatectomy, a significant decrease in 2n (1 diploid nucleus) hepatocytes and a significant increase in 8n (1 octoploid nucleus) hepatocytes occurred. In contrast, 24 h following induction of acute hepatic failure by single D-galactosamine or thioacetamide injections, a significant increase in 2n hepatocytes was observed, whereas the proportion of 8n hepatocytes remained unchanged. The liver ploidy returned to basal values within 21 days in all cases. In cirrhotic livers induced by chronic thioacetamide injections, the rate of 2n hepatocytes was about ten times that of the controls having the same age, while 4n (1 tetraploid nucleus) and 8n hepatocytes were one third of controls. The binucleation rate was also significantly decreased.

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Glycogen content in hepatocytes is related with their size in normal rat liver but not in cirrhotic one

et al. 2016

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Hepatocytes differ from one another by the degree of the ploidy, size, position in the liver lobule, and level of the DNA-synthetic processes. It is believed, that the cell size exerts substantial influence on the metabolism of the hepatocytes and the glycogen content in them. The aim of the present study was to test this hypothesis. Dry weight of hepatocytes, their ploidy and glycogen content were determined in the normal and the cirrhotic rat liver. Liver cirrhosis in rats was produced by chronic inhalation of CCl 4 vapours in the course of 6 months. A combined cytophotometric method was used. Dry weight of the cell, its glycogen and DNA content were successively measured on a mapped preparation. Hepatocytes of each ploidy class in the normal and the cirrhotic rat liver accumulated glycogen at the same rate. In the normal liver, there was a distinct correlation between the size of hepatocytes and glycogen content in them. This correlation was observed in each ploidy class, and was especially pronounced in the class of mononucleate tetraploid hepatocytes. In the cirrhotic liver, there was no correlation between the size of the cells and their glycogen content. The impairment of liver lobular structure probably explains the observed lack of correlation between hepatocyte size and their glycogen content in the cirrhotic liver. Key terms hepatocytes; glycogen; dry weight of hepatocytes; ploidy; liver cirrhosis GLUCOSTATIC activity is a major function of the liver. Hepatocytes contribute to the sustenance of a constant glucose level in the blood by synthesizing glycogen from glucose after a meal and breaking it down when necessary. Glycogenesis and glycogenolysis, ensured by several enzymes, are regulated by hormonal, nervous, substrate and other mechanisms (1,2) as well as tissue and cell factors (3,4).The population of hepatocytes is not homogeneous. These cells differ not only in their specific functional activity but also in ploidy level, size, position in the liver lobule, level of proliferative activity etc. (5-7). Glycogen content and enzymatic activity of hepatocytes are known to be determined by their location in the liver lobule, the ploidy level and the cell cycle phase (6,(8)(9)(10)(11). However, we know almost nothing about the relations between the functional activity of hepatocytes, including glycogen content and accumulation rate, and their size.The size of hepatocytes may considerably vary. Even hepatocytes of the same ploidy class may differ by several times as to volume and dry weight (12,13). The reasons of this variability are not quite clear. At the same time, the size of hepatocytes is crucial for their metabolism.It has been suggested that there is a certain relation between the size of hepatocytes and the glycogen content in them: larger cells are richer in glycogen (14). The aim of the present study was to test this hypothesis. The aim of the present study was

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Hepatocyte ploidy in normal young rat

Cited by 37 publications

References 31 publications

Embryonic Stem Cells Contribute to Mouse Chimeras in the Absence of Detectable Cell Fusion

Embryonic Stem Cells Contribute to Mouse Chimeras in the Absence of Detectable Cell Fusion

Hepatocyte Ploidy in Regenerating Livers after Partial Hepatectomy, Drug-Induced Necrosis, and Cirrhosis

Glycogen content in hepatocytes is related with their size in normal rat liver but not in cirrhotic one

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