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PubMed1980

Two-dimensional gel electrophoresis of nuclear phosphoproteins of Novikoff hepatoma and regenerating liver.

Two-dimensional polyacrylamide gel electrophoresis patterns of 32P-labeled nuclear proteins in Novikoff hepatoma and regenerating liver relatively small number of the many nuclear protein spots were phosphorylated. The major phosphoprotein spots differed in size and shape from the stained protein spots. Seven phosphoproteins, 125/5.8, 125/7.2, 100/5.9, 85/5.9, 56/5.1, and 50/5.1 (mol. wt. x 10-(3)/pI in the Novikoff hepatoma nuclei were not found in the 18-h regenerating liver nuclei. One nuclear phosphoprotein, 54/6.5, was found in the liver but not in the hepatoma. The four major phosphoproteins, 45/5.3-6.0, 40/5.3-6.0, 35/5.3-6.0, and 25/5.8-6.8 containing about 60% of the total 32Pi of the nuclear proteins, were found in both types of cells. These proteins were of unusual density and shape. Some proteins, such as 125/5.8 and 85/5.9, were enriched in 0.01 M tris extract of the tumor, and proteins 125/7.2 and 56/6.1 were enriched in 0.35 M NaCl extract of the tumor.

Animals↗
PubMed2005

A novel molecular marker of pituitary tumor transforming gene involves in a rat liver regeneration.

BACKGROUND: Pituitary tumor transforming gene (PTTG), homologous to a mammalian securin, plays a pivotal role in cell transformation, however, its biological function(s) in normal tissues is not fully understood. Because the liver is a regenerative organ, the relevant biological function of PTTG in the liver would be more feasible to understand PTTG. Also, PTTG may be involved in the liver regeneration. MATERIALS AND METHODS: Expressions of rat hepatic PTTG messengerRNA (mRNA) and cellular immunoreactivities during the cell proliferative period of the liver regeneration both in vitro and in vivo were tested. RESULTS: PTTG expression of the rat primary hepatocyte was stimulated by HGF in a dose dependent manner, and was suppressed when hepatocyte proliferation was inhibited by transforming growth factor-beta1. A positive PTTG immunoreactive co-localizing with 5-bromo-2'-deoxyuridine (BrdU) in the hepatocyte nucleus was found and there was a concurrent sister chromatin itself by the immunofluorescent labeling of PTTG with cytokeratin 18 (CK18). DISCUSSION: Since the correlation of PTTG mRNA expression, cell proliferation and immunoreactivity were observed in primary rat cultured hepatocytes, PTTG may be a novel marker of cell proliferation both in vitro and in vivo liver regeneration.

Animals↗
PubMed2003

The molecular physiology of liver regeneration following partial hepatectomy.

The ability of the liver to regenerate after resection has been known for many years. Two reports from Germany in the late 1800s probably mark the introduction of the phenomenon into the scientific literature, but in the early 1900s the first reviews of this subject had appeared in the English literature. Predating these early scientific reports the legends from the Greek mythology described the fate of Prometheus. As punishment for defying Zeus and revealing the secret of fire to man, Prometheus was chained to a rock and each day had part of his liver ripped out by an eagle which, returning the following day, repeated the torture because his liver regenerated itself overnight. Although the speed of regeneration in the Greek legend is somewhat greater than that observed either clinically or in the laboratory, the myth does serve to emphasise the remarkable ability of the liver to repeatedly regenerate following repeated resections. This review aims to summarise the more recent literature concerning the early molecular events accompanying liver regeneration and to integrate this with the existing knowledge of this subject.

Animals↗
PubMed1999

Levels of hepatic stimulator substance in liver regenerating process of partially hepatectomized rats pretreated with a single dose of carbon tetrachloride.

Liver regeneration after injury with carbon tetrachloride (CCl4) followed by partial hepatectomy is a complex model involving toxicological, inflammatory, and necrotic processes. In the present study, the time-course of hepatic regenerative process was investigated in relation to hepatic stimulator substance (HSS) activity, administration of a single dose of CCl4 and partial (70%) hepatectomy in male rats. To evaluate liver injury events, the levels of serum aspartic aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were measured. Hepatic DNA synthesis reached a maximum at 36 hr after hepatectomy in contrast to the reported 24-hr and 32-hr peaks observed in nontreated hepatectomized rats. On the other hand, HSS activity appeared to peak at 28, 40, and 44 hr after hepatectomy in CCl4-treated rats, and it was quite a lot lower at 24, 32, 36, 48, and 60 hr. The hypothesis that HSS promotes liver regeneration but it does not initiate it, as other factors have been found to do, is discussed.

Alanine Transaminase↗
PubMed1993

Quantitative and/or qualitative changes in the p21-H-ras post-translational products in regenerating liver and during hepatocarcinogenesis.

Using Western immunoblot analysis with the Ras-10 monoclonal antibody (MAb), we characterized 4 post-translational products of the c-H-ras gene in rat tissues, and showed that they were readily distinguished from the normal p21-N-ras or Val12 mutant p21-H-ras products. In the present study, we used this approach to compare the electrophoretic pattern of p21-H-ras during chemically induced hepatocarcinogenesis in rats with that in control liver samples. Three types of pattern were defined in the liver samples taken at various stages of malignant progression. The first type, like all the 18 normal samples analyzed, was characterized by a predominant intensity of spot a (corresponding to the palmitoylated p21-H-ras product). This was observed in all the samples at the stage of foci and only in a proportion of nodular or tumoral tissues. The second type of pattern deviates from this normal basal pattern by a higher relative level in one or more of the precursors of the fully processed p21-H-ras product. It was observed in all liver samples at the stage of nodules and in a proportion of tumors, but also in all samples from fetal or regenerative liver, thus suggesting an association with high proliferative activity of the hepatocytes. The third type of pattern was characterized by the presence of spots never detected in any of the normal rat tissue or cells that we investigated. Abnormal spots were observed in nodular liver samples and in hepatocarcinomas, indicating that they probably correspond to mutant p21-H-ras products. The fact that the 2 types of abnormal products were not constantly associated with neoplasms indicates that if they play a role in their induction or maintenance this may also be achieved in an independent way, perhaps, but not necessarily, involving another mutation of the ras gene.

Animals↗
PubMed1978

Polyamine metabolism in regenerating livers from normal and hypocalcemic rats.

There is a marked increase in the concentration of putrescine during the first ten hours following partial hepatectomy in rats. The concentration of spermidine also increases but to a smaller degree. Putrescine levels return to normal between 10 and 24 hours after the operation, whereas the increased spermidine level is maintained. The production of putrescine and spermidine appears to be initiated by the induction of ornithine decarboxylase which shows a single peak of activity at four hours after hepatectomy. The activity of S-adenosylmethionine decarboxylase shows little change following hepatectomy. The changes in polyamine levels and the activities of the enzymes of polyamine metabolism are not affected by thyroparathyroidectomy 72 hours prior to hepatectomy. Thus although these hypocalcemic conditions considerably reduce and delay DNA synthesis and mitosis, the prereplicative changes in polyamine metabolism still occur. These data suggest that the hepatocytes in hypocalcemic animals have become activated and moved to an advanced stage of prereplicative development before being blocked.

Adenosylmethionine Decarboxylase↗
PubMed1981

Regulation of the prereplicative changes in the synthesis and transport of messenger and ribosomal RNA in regenerating livers of normal and hypocalcemic rats.

Partial hepatectomy induces increases in the synthesis of both messenger (poly(A)+) and ribosomal (poly(A)-) RNA, which precede the initiation of DNA synthesis. The increase in poly(A)+RNA, which commences soon after surgery and reaches a peak 1-3 hours later is particularly striking. Disruption of this early increase in poly(A)+RNA synthesis by colchicine (and other microtubule disrupters) or indomethacin results in a failure to initiate DNA synthesis. This suggests that prostaglandins and the microtubules are involved in the mechanism of proliferative activation. Hypocalcemia, which also prevents the initiation of DNA synthesis, has no effect on the prereplicative changes in either messenger or ribosomal RNA synthesis. These results help to define the critical stages of prereplicative development and give some insight into their regulation.

Animals↗
PubMed1985

Erythropoietin production by macrophages in the regenerating liver.

The site of erythropoietin (Ep) production and/or storage in the rat liver was determined. A guinea pig anti-Ep was produced against purified rat Ep (64,096 +/- 4j064 IU/mg). This antibody was found to be highly specific using rocket immunoelectrophoresis, Ouchterlony gel diffusion methods, and immunoprecipitin reactions as well as Ep neutralization tests (capable of completely neutralizing up to 2,000 IU Ep/mg). This anti-Ep was labeled with either fluorescein for light microscopic study or ferritin for electron microscopy. Kupffer cells showed varying degrees of labeling after hepatectomy alone or hepatectomy combined with nephrectomy and/or hypoxia. Greatest labeling was seen in Kupffer cells of rats that were nephrectomized 48 hr posthepatectomy and kept at ambient pressure. No labeling of hepatocytes or vascular and bile duct endothelium was noted.

Animals↗
PubMed1994

Influence of transcriptional regulation and mRNA stability on hemopexin gene expression in regenerating liver.

The hepatic response to systemic injury is characterized by alterations in the synthesis of plasma proteins, while acute injury to the liver can lead to rapid proliferation of hepatocytes. The hemopexin gene was found to be markedly induced in rat liver following both sham surgery (SS) and 70% partial hepatectomy (PH), models of systemic injury and hepatic proliferation, respectively. Transcriptional and post-transcriptional regulation of this gene was evaluated to examine the mechanisms of hemopexin mRNA expression in these models. Significant transcriptional activation was observed within 6 h of either surgery, with a more pronounced effect after PH. In both processes, transcription rates returned to baseline values by 24 h after surgery, although marked elevations in mRNA steady-state levels were noted for at least 72 h. At each time point, levels of hemopexin mRNA were more abundant following PH than after SS, in part due to greater transcriptional induction. In addition, posttranscriptional mechanisms appeared to contribute to the increased expression of hemopexin post-PH. The in vivo half-life of the 1.6-kb hemopexin transcript was determined to be considerably greater than 12 h in control, sham-operated, and PH animals. The exceptionally long mRNA half-life appears to be an important but complex factor in the kinetics of hemopexin gene regulation.

Animals↗
PubMed1997

Cell-cycle regulated expression of Rap1 in regenerating liver.

Rap1 proteins are capable of competing with Ras p21 for binding to effectors, and of antagonizing some Ras-induced effects, but their participation in normal growth regulation has not been established. The level of Rap1 protein and the expression of the rap1A gene were examined by immunoblotting and Northern analysis during the regenerative growth response in rat liver following partial hepatectomy. Protein and mRNA were significantly down-regulated prior to and during the onset of DNA synthesis. The timing of this effect is consistent with a model in which expression of Rap1 is turned off or down to allow the initiation of proliferation.

Animals↗
PubMed2002

Nuclear translocation of mouse polycomb m33 protein in regenerating liver.

Immunoblots probed with an antibody to M33 protein, a homolog of Drosophila Polycomb, revealed that most M33 in adult mouse liver had a higher electrophoretic mobility than that in F9 embryonal carcinoma cells. High-mobility 60-kDa M33 localized in the cytoplasmic fraction of liver homogenates, and two less abundant 66- and 70-kDa species were detected in the nuclear fraction. Immunocytochemistry of freeze-substituted tissues showed a punctate pattern of immunofluorescence in the cytoplasm of hepatic parenchymal cells. Nuclear M33 isoforms treated with alkaline phosphatase had increased mobilities corresponding to cytoplasmic M33. In partially hepatectomized mice, nuclear M33 isoforms appeared after 48 h, near the time of maximum DNA synthesis as measured by bromodeoxyuridine incorporation. By 60 h, most M33 was in the form of these low-mobility species, and the pattern of immunofluorescence suggested the existence of chromatin-bound and free states of the protein in the nucleus. Thereafter, high-mobility 60-kDa M33 reappeared. The data are consistent with a phosphorylation-associated translocation mechanism that is a cell cycle-dependent.

Active Transport, Cell Nucleus↗