Figure 1.
Scheme of enzymatic reactions involved in basic liver functions and metabolism.
(A) Function of key enzymes of nitrogen metabolism and their zonation in healthy liver hepatocytes. (B) Enzymatic reactions involved in glucose storage and release in hepatocytes. (C) Metabolic activation of CCl4 in pericentral hepatocytes.
Figure 2.
In situ hybridization for genes from nitrogen metabolism.
In situ hybridization of mouse liver sections with probes for selected genes involved in nitrogen metabolism at different time points post CCl4 injection. In each panel, genes were visualized by dual staining with yellow and violet dye precipitation. Gene names are indicated on the left in the respective color. Co-staining for both genes in the same area resulted in dark “brown” staining. Pictures were captured with 4x objective.
Figure 3.
In situ hybridization for genes from carbohydrate metabolism.
In situ hybridization of mouse liver sections with probes for selected genes of carbohydrate metabolism at different time points post CCl4 injection. Genes were visualized by dual staining with yellow and violet dye precipitation. Gene names are indicated on the left in the respective color. Co-staining for both genes in the same area resulted in dark “brown” staining. Pictures were captured with 4x objective.
Figure 4.
Higher resolution in situ hybridization images.
In situ hybridization of mouse liver sections from untreated animals and form days 1 to 3 after CCl4 injection, with higher magnification (20x objective). Genes analyzed are indicated at the left in the respective color for each row. Co-staining for both genes in the same area resulted in dark “brown” staining. Specific areas of the liver tissue (acini) are marked: central vein (cv), portal vein/area (pv).
Figure 5.
Quantitative analysis of over all gene expression levels.
Analysis of total gene expression levels by reverse transcription quantitative real time PCR using primer pairs recognizing the following genes: arginase (Arg1), glycogen synthase (Gys2), glucagon receptor (Gcgr), Gapdh, glutaminase (Gls2), glutamine synthase (Gs), glucose-6-phosphatase (G6pc), cytochrom p450 2E1 (Cyp2e1). Albumin (Alb) was used for normalization.
Figure 6.
Graphical summary of the area specific gene expression patterns.
Results from the ISH analysis are summarized into 4 conditions. Untreated liver (normal), and after 3h and 6h, 2 days (2d) and 3 days (3d) upon CCl4 treatment. Intensity of the color reflects the relative intensity of gene expression for each gene (intense/dark color: strong/high signal). The intensities only visualize the relative distribution of the respective mRNA along the axis from the periportal area to the pericentral vein (blue: higher periportal: green; higher pericentral). Open cells (circular light to dark coloring) are used to indicate non homogenous speckled patterns observed in the ISH images.