Lobule-dependent zonal measurement (LZM) method for the determination of cell proliferation in the liver

Bahnemann, Rainer; Mellert, W.

doi:10.1016/s0940-2993(97)80006-1

Cited by 17 publications

(13 citation statements)

References 29 publications

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“…Ten to 15 fields were counted. The labeling indices of hepatocytes were determined in different zones as described previously (Bahnemann and Mellert, 1997). To guarantee lobule comparison, the distance from the portal tract to the central vein was determined.…”

Section: Methodsmentioning

confidence: 99%

Overlapping Transcriptional Programs Regulated by the Nuclear Receptors Peroxisome Proliferator-Activated Receptor α, Retinoid X Receptor, and Liver X Receptor in Mouse Liver

Anderson

Dunn²,

Laughter³

et al. 2004

Mol Pharmacol

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Lipid homeostasis is controlled in part by the nuclear receptors peroxisome proliferator (PP)-activated receptor ␣ (PPAR␣) and liver X receptor (LXR) through regulation of genes involved in fatty acid and cholesterol metabolism. Exposure to agonists of retinoid X receptor (RXR), the obligate heterodimer partner of PPAR␣, and LXR results in responses that partially overlap with those of PP. To better understand the gene networks regulated by these nuclear receptors, transcript profiles were generated from the livers of wild-type and PPAR␣-null mice exposed to the RXR pan-agonist 3,7-dimethyl-6S,7S-methano, 7-[1,1,4,4-tetramethyl-1,2,3,4-tetrahydronaphth-7-yl]-2E,4E-heptadienoic acid (AGN194,204) or the PPAR pan-agonist WY-14,643 (WY; pirinixic acid) and compared with the profiles from the livers of wild-type and LXR␣/LXR␤-null mice after exposure to the LXR agonist N- (2,2,2-trifluoroethyl)-N-[4-(2,2,2-trifluoro-1-hydroxy-1-trifluoromethylethyl)phenyl] sulfonamide (T0901317). All 218 WY-regulated genes altered in wild-type mice required PPAR␣. Remarkably, ϳ80% of genes regulated by AGN194,204 required PPAR␣ including cell-cycle genes, consistent with AGN-induced hepatocyte proliferation having both PPAR␣-dependent and -independent components. Overlaps of ϳ31 to 62% in the transcript profiles of WY, AGN194,204, and T0901317 required PPAR␣ and LXR␣/LXR␤ for statistical significance. Ofthe 50 overlapping genes regulated by T0901317 and WY, all but one were regulated in a similar direction. These results 1) identify new transcriptional targets of PPAR␣ and RXR important in regulating lipid metabolism and liver homeostasis, 2) illustrate the importance of PPAR␣ in regulation of gene expression by a prototypical PP and by an RXR agonist, and 3) provide support for an axis of PPAR␣-RXR-LXR in which agonists for each nuclear receptor regulate an overlapping set of genes in the mouse liver.

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Section: Methodsmentioning

confidence: 99%

Overlapping Transcriptional Programs Regulated by the Nuclear Receptors Peroxisome Proliferator-Activated Receptor α, Retinoid X Receptor, and Liver X Receptor in Mouse Liver

Anderson

Dunn²,

Laughter³

et al. 2004

Mol Pharmacol

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“…The mean distance between these vessels was ∼350 µm. The fields were subdivided into 3 equal parts (∼100 hepatocytes) corresponding to the lobule-dependent zonal measurement method (LZM) (Bahnemann and Mellert, 1997): periportal (zone 1), midzonal (zone 2), and perivenous (zone 3). Total LI was calculated as an average of all 3 zones.…”

Section: Evaluation Schemementioning

confidence: 99%

Zonal Distribution of Cell Proliferation in the Liver of Untreated B6C3F1 and C57BL Mice

et al. 2004

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To determine the lobular distribution of hepatocellular proliferation, S-phase response was measured in untreated adult male B6C3F1 and C57BL mice at ages 11, 14, and 23 weeks. The percentage of cells in S-phase (labeling index, LI) was evaluated using immunohistochemical detection of 5-bromo-2 -deoxyuridine (BrdU). The BrdU was delivered either by a single ip injection 2 hr prior to sacrifice or via an osmotic minipump implanted subcutaneously for 3 or 7 days. Mitotic and apoptotic indices were determined on H&E stained sections. Animals of both strains and all ages revealed highest LI in the intermediate compartment of the liver acinus (zone 2) irrespective of the length of BrdU application. Cell proliferation in this zone was at all time points significantly higher than elsewhere in the liver. The mitotic index confirmed the data obtained by the S-phase response study. Apoptosis was rarely observed. With regard to rodent data in chemical carcinogenesis and the way they should be evaluated, this study proved that the acinar distribution of proliferating cells in liver of mice is different from that in rats, since, according to the literature, rats reveal highest cell proliferative activity in the periportal zone.

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“…Because liver regeneration is inhomogeneous within a liver lobe, but also between liver lobes, the selection and the total size of the ROIs to be analyzed can also influence the result of the BrdU-LI on the postanalytical level. Bahnemann and Mellert (1997) described the LZM method for determination of cell proliferation in the liver. According to the rules of the LZM method, measurement fields should be of equal size.…”

Section: Discussionmentioning

confidence: 99%

“…The total size of the ROI to be analyzed was determined by both the classical statistical approach based on the central limit theorem and the lobule-dependent zonal measurement (LZM) method (Bahnemann and Mellert 1997). A coefficient of variation (CV) of below 10% was considered to be acceptable, inasmuch as the biological variation between individual animals was much higher and can reach more than 30%, as already shown by Fabrikant as early as 1968 (Fabrikant 1968;Schmitz and Hof 2000).…”

Section: Estimation Of the Total Size Of The Roi To Be Analyzedmentioning

confidence: 99%

Statistical and Economical Efficiency in Assessment of Liver Regeneration Using Defined Sample Size and Selection in Combination With a Fully Automated Image Analysis System

Deng

Kleinert

Huang

et al. 2009

J Histochem Cytochem.

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Quantification of liver regeneration is frequently based on determining the 5-bromo-2-deoxyuridine labeling index (BrdU-LI). The quantitative result is influenced by preanalytical, analytical, and postanalytical variables such as the region of interest (ROI). We aimed to present our newly developed and validated automatic computer-based image analysis system (AnalySIS-Macro), and to standardize the selection and sample size of ROIs. Images from BrdU-labeled and immunohistochemically stained liver sections were analyzed conventionally and with the newly developed AnalySIS-Macro and used for validation of the system. Automatic quantification correlated well with the manual counting result (r=0.9976). Validation of our AnalySIS-Macro revealed its high sensitivity (>90%) and specificity. The BrdU-LI ranged from 11% to 57% within the same liver (32.96 +/- 11.94%), reflecting the highly variable spatial distribution of hepatocyte proliferation. At least 2000 hepatocytes (10 images at 200x magnification) per lobe were required as sample size for achieving a representative BrdU-LI. Furthermore, the number of pericentral areas should be equal to that of periportal areas. The combination of our AnalySIS-Macro with rules for the selection and size of ROIs represents an accurate, sensitive, specific, and efficient diagnostic tool for the determination of the BrdU-LI and the spatial distribution of proliferating hepatocytes.

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Lobule-dependent zonal measurement (LZM) method for the determination of cell proliferation in the liver

Cited by 17 publications

References 29 publications

Overlapping Transcriptional Programs Regulated by the Nuclear Receptors Peroxisome Proliferator-Activated Receptor α, Retinoid X Receptor, and Liver X Receptor in Mouse Liver

Overlapping Transcriptional Programs Regulated by the Nuclear Receptors Peroxisome Proliferator-Activated Receptor α, Retinoid X Receptor, and Liver X Receptor in Mouse Liver

Zonal Distribution of Cell Proliferation in the Liver of Untreated B6C3F1 and C57BL Mice

Statistical and Economical Efficiency in Assessment of Liver Regeneration Using Defined Sample Size and Selection in Combination With a Fully Automated Image Analysis System

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