Quantitative analysis of cell allocation during liver development, using the spf(ash)-heterozygous female mouse.

Mosaicism of ornithine transcarbamylase (OTC) expression in hepatocytes was quantitatively analyzed during liver development of the spf(ash)-heterozygous female mouse. Because the mosaic patterns depend on cell migration and cell mixing, such analysis could give insights on the growth pattern or allocation pattern of hepatocytes during liver development. Complex mosaic patterns of OTC-positive and -negative hepatocytes were observed in sections of fetal and postnatal livers. Sizes of patches, which were aggregates of OTC-positive or -negative hepatocytes, increased during development. Patches were slender and comparatively simple in 15.5- and 17.5-day fetal and neonatal livers. Quantitative analysis of patch shapes demonstrated that undulation of patches was maximal at 7 postnatal days. Patches with nodular shapes also started to increase in number at this stage. Isolated patches in sections of fetal livers and postnatal livers three-dimensionally connected with one another. However, especially in fetal livers, in which OTC-positive patches were minor, due to the presence of abundant hemopoietic cells, isolated three-dimensional patches consisting of approximately 5 to 70 cells were often found. They were shaped like slender branching or zigzag-shaped cords, but no definite orientation such as portal-central was observed in them at any stage. These results suggest that hepatocytes contiguously allocate their daughter cells as zigzag-shaped or branching cords at younger stages. Some hepatocytes grow with nodular formation after 7 postnatal days. Migration and mixing of hepatocytes appear to be more extensive at fetal stages than in the adult liver. Immunohistochemical analysis of intercellular junction proteins (E-cadherin, connexins 26 and 32, occludin, and ZO-1) also revealed that their expression and distribution changed in hepatocytes during development, which may be correlated with the OTC mosaic patterns.