Comparison of Proliferating Cell Nuclear Antigen to Tritiated Thymidine As a Marker of Proliferating Hepatocytes in Rats

Foley, Julie F.; Ton, Thai-Vu; Maronpot, Robert R.; Butterworth, Byron E.; Goldsworthy, Thomas L.

doi:10.2307/3431868

Cited by 27 publications

(32 citation statements)

References 16 publications

(16 reference statements)

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“…Previous studies have demonstrated that immunohistochemical staining patterns of PCNA, using paraffin-embedded liver tissue sections, are comparable to tritiated thymidine incorporation in rodents, suggesting that the nucleus alone shows positive reactivity in G1 phase and strong positive reactivity in S phase, but neither the nucleus nor the cytoplasm of hepatocytes shows a positive reaction in G0 phase (Foley et al 1993). In the present study, the labeling index of BrdU, analogous to tritiated thymidine, and the proportion of PCNA-stronglypositive hepatocytes were equally reduced in DR in vivo, while the proportion of neither PCNA-positive hepatocytes nor TUNEL-positive hepatocytes was influenced 366 Fig.…”

Section: Discussionmentioning

confidence: 95%

“…Immunohistochemistry for bromodeoxyuridine (BrdU) (Shimokawa et al 1997) and proliferating cell nuclear antigen (PCNA) (Foley et al 1993;Higami et al 1997a) was performed to evaluate the proliferation of hepatocytes. Immunohistochemistry for bromodeoxyuridine (BrdU) (Shimokawa et al 1997) and proliferating cell nuclear antigen (PCNA) (Foley et al 1993;Higami et al 1997a) was performed to evaluate the proliferation of hepatocytes.…”

Section: Introductionmentioning

confidence: 99%

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Dietary restriction reduces hepatocyte proliferation and enhances p53 expression but does not increase apoptosis in normal rats during development

Higami

Shimokawa

Ando

et al. 2000

Cell and Tissue Research

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Dietary restriction (DR) retards physical growth, resulting in small body size, reduced liver weight and reduced number of hepatocytes in rats. We examined the effects of DR on proliferation and apoptosis of hepatocytes during development and explained these changes subcellularly using immunohistochemistry for bromodeoxyuridine (BrdU) incorporation, proliferating cell nuclear antigen (PCNA) and p53, terminal dUTP nick end labeling (TUNEL) and quantitative reverse transcription-polymerase chain reaction (RT-PCR) for TGFalpha, TGFbeta1, p53, Fas and TNF receptor (TNFr). Tissue samples included the livers of 3-month-old male Fischer 344 rats fed ad libitum (AL) or on 70% DR. DR significantly reduced the proportions of BrdU-positive and PCNA-strongly-positive hepatocytes compared with AL rats but not the proportions of PCNA-positive hepatocytes and TUNEL-positive hepatocytes. On the other hand, DR enhanced the expression of p53 and Fas mRNAs but failed to influence the expression of TGFalpha, TGFbeta1 and TNFr mRNAs. Moreover, DR significantly increased the proportion of p53-positive hepatocytes. Our findings suggest that DR suppresses the proliferation of hepatocytes, resulting in a reduced number of hepatocytes during development. This process may be mediated by overexpression of p53 suppressor gene. While DR accelerates the expression of Fas antigen, this result does not influence the rate of apoptosis of hepatocytes under physiological conditions.

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Dietary restriction reduces hepatocyte proliferation and enhances p53 expression but does not increase apoptosis in normal rats during development

Higami

Shimokawa

Ando

et al. 2000

Cell and Tissue Research

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“…PCNA staining was performed according to a modification of the protocol of Foley et al (5). Briefly, the slides were dried overnight, deparaffinized through three changes of xylene, hydrated to distilled water, and incubated with PC-10 antibody (Novocastra Laboratory, Newcastle-on-Tyne, UK) in capillary gap slides at 35°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Effects of Dietary Soybean Estrogens on the Reproductive Tract in Female Rats

Tansey

Hughes

Cline

et al. 1998

Experimental Biology and Medicine

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The estrogenic actions of dietary phytoestrogens have raised concerns regarding the potential DES-like developmental effects on the female genital tract, but the growing evidence of cardioprotective benefits of dietary soybean estrogens provides the impetus to assess the effects of these compounds in adult female models of the menopause. We conducted an experiment in ovariectomized rats to determine the independent effects of dietary soybean estrogens (SBE) and the interactions of these agents with the commonly used pharmaceutical estrogen preparation (conjugated equine estrogens, CEE) in the vagina and uterus. We looked at the effects of SBE and CEE, alone and in combination, on uterine weight, body weight, vaginal cytology, uterine luminal epithelial height, and immunohistochemical staining for proliferating cell nuclear antigen (PCNA), lactoferrin (Ltf), and apoptosis. Ovariectomized rats were fed diets containing casein or soybean protein (SBE, low dose = 11.6 mg isoflavones/ 1800 cal; high dose = 117.8 mg/1800 cal), with no CEE, low dose CEE (0.313 mg/1800 cal), or high dose CEE (0.625 mg/1800 cal) added. In this study, SBE did not demonstrate estrogenic activity for uterine weight or vaginal cytology. We also found no estrogenic effects of these doses of SBE for PCNA, apoptosis, Ltf staining, or for LEH measurements. In addition, our results regarding the interactions of SBE and CEE do not show any evidence that the combination is additive in effect. On the contrary, the LEH response induced by low levels of CEE, was reduced by high levels of SBE. Furthermore, the Ltf response induced by CEE also was reduced by high levels of SBE. This suggests that high doses of SBE may antagonize the estrogen-agonist actions of low doses of CEE in the rat uterus. Our results in the ovariectomized rat model of menopause suggest that dietary soybean estrogens will not elicit a pattern of effects that simply recapitulates those of steroidal estrogens.

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“…A labeling index was obtained by counting 1000 hepatocytes/rat. Additional sections were stained immunohistochemically for the endogenous replication marker, proliferating cell nuclear antigen (PCNA) [10,11], using a PCNA monoclonal antibody (Coulter Corporation, Hialeah, FL) and an antimouse IgM biotinylated secondary antibody (Vector Laboratories) followed by streptavidin and 3,3-diaminobenzidine. A proliferating index was obtained based on 1000 hepatocytes/rat.…”

Section: Assessment Of Hepatocellular Proliferationmentioning

confidence: 99%

Two‐dimensional electrophoresis of liver proteins: Characterization of a drug‐induced hepatomegaly in rats

et al. 2000

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Two-dimensional electrophoresis (2-DE) of liver proteins was applied to further characterize an unusual drug-induced increase in hepatocellular rough endoplasmic reticulum (RER) in Sprague-Dawley rats given a substituted pyrimidine derivative. Absolute liver weights of drug-treated rats (9.9 +/- 0.4 g) increased above vehicle-treated controls (7.2 +/- 0.2 g) by 37%. Light microscopy revealed diffuse granular basophilia of the hepatocellular cytoplasm, uncharacteristic of hepatocytes and suggested cells rich in ribosomes, which was confirmed by electron microscopy. Immunostaining for cell proliferation, viz., 5-bromo-2'-deoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA), indicated marked hepatocellular proliferative activity. 2-DE of solubilized liver using an ISO-DALT gel system indicated significant (p<0.001) quantitative changes in at least 17 liver proteins (12 increased, 5 decreased) compared to controls. The protein with the largest increase was homologous to acute-phase reactant, contrapsin-like protein inhibitor-6. Other markedly upregulated proteins were methionine adenosyltransferase, a catalyst in methionine/ATP metabolism and mitochondrial HMG-CoA synthase, involved in cholesterol synthesis. The complementary strategies of 2-DE coupled either with database spot mapping or protein isolation and amino acid sequencing successfully identified a subset of proteins from xenobiotic-damaged rodent livers, the expression of which differed from controls. However, the current bioinformatics platform for rodent hepatic proteins and limited knowledge of specific protein functionality restricted application of this proteomics profile to further define a mechanistic basis for this unusual hepatotoxicity.

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Comparison of Proliferating Cell Nuclear Antigen to Tritiated Thymidine As a Marker of Proliferating Hepatocytes in Rats

Cited by 27 publications

References 16 publications

Dietary restriction reduces hepatocyte proliferation and enhances p53 expression but does not increase apoptosis in normal rats during development

Dietary restriction reduces hepatocyte proliferation and enhances p53 expression but does not increase apoptosis in normal rats during development

Effects of Dietary Soybean Estrogens on the Reproductive Tract in Female Rats

Two‐dimensional electrophoresis of liver proteins: Characterization of a drug‐induced hepatomegaly in rats

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