Takeo Mizuno

Differentiation of functional hepatocytes and biliary epithelial cells from immature hepatocytes of the fetal mouse in vitro

Differentiation of functional hepatocytes and biliary epithelial cells from immature hepatocytes was analysed in vitro. When fetal mouse liver fragments containing immature hepatocytes but no bile ducts were cultured organotypically, the immature hepatocytes differentiated into large hepatocytes. Some of these expressed bile duct markers such as cytokeratin and Dolichos biflorus agglutinin-binding sites, though only to a small extent, and typical intrahepatic bile duct cells failed to differentiate. Dexamethasone stimulated immature hepatocytes to differentiate into both mature hepatocyte and biliary epithelial cell lineages. Especially in the liver fragments cultured on Matrigel, dexamethasone stimulated the expression of bile duct markers (such as cytokeratin and binding sites for two types of lectin) in the immature hepatocytes. These results support the idea that immature hepatocytes can differentiate into both mature hepatocytes and biliary epithelial cells during normal development of the mouse liver, and suggest that glucocorticoids stimulate both these differentiation pathways. It also seems that basal laminar components may play a role in bile duct differentiation.

Cytokeratin Expression in the Stomach Epithelia of the Chicken Embryo is Regulated by Epithelial-Mesenchymal Interactions

Abstract Regulatory mechanisms of cytokeratin expression in the stomach (the proventriculus and the gizzard) epithelia of chicken embryos were examined by epithelial-mesenchymal recombination experiments. In the proventriculus, antigens detected by monoclonal antibodies (PKK1 and AE3) against cytokeratins invariably decreased in gland cells which synthesize pepsinogen, while luminal and duct epithelial cells were positive to these antibodies. When recombined and cultured with the proventricular mesenchyme, the esophageal, proventricular and gizzard epithelia formed proventriculus-type, pepsinogen-expressing glands and the expression patterns of PKK1 and AE3 antigens were identical to that in the normal proventriculus. Under the same experimental conditions, the small-intestinal and allantoic epithelia also formed complex glands but did not express pepsinogen, and PKK1 and AE3 antigens were actively expressed throughout the epithelia. These results indicate that the proventricular mesenchyme can regulate cytokeratin expression and that the expression of cytokeratins in epithelial cells is closely related to the physiological characteristics of cells such as pepsinogen production.

Epithelial Metaplasia Induced Amnionic Ectoderm by the Dermis of Chicken Embryos1

Amnionic ectoderm of 6.8‐day chicken embryos was associated with 6.8‐day dorsal dermis or 13–15‐day scale dermis and cultured on host chorio‐allantoic membrane for 8 days. The amnionic ectoderm, recombined and cultured with the dorsal dermis, developed feather filaments consisted of a feather root, a horny sheath, and barb ridges. With several feather keratin‐specific monoclonal antibodies (4E12 and 1F3), these structures in the induced feather filaments were shown to express feather‐specific keratin antigens.

Demonstration of sucrase immunoreactivity of the brush border induced by duodenal mesenchyme in chick stomach endoderm

SummaryWhen stomach endoderm of chick embryos was recombined and cultured with duodenal mesenchyme, the endoderm developed a brush border structure over a large area and also differentiated into mucous cells in a small area according to its own developmental fate. In the present investigation, we examined whether the induced brush border structure expressed sucrase antigen by immunoelectron microscopy using the antiserum raised against chicken sucrase. Sucrase immunoreactivity could be detected as ferritin particles in the region where the brush border was induced, whereas it was never detected on microvilli of endodermal cells which differentiated into the mucous cells. Thus, almost all of the endodermal cells could be identified as either small intestine-type cells possessing the sucrase antigen or stomach-type cells possessing mucous granules but not the sucrase antigen. The results indicate that stomach endodermal cells of chick embryos can differentiate not only morphologically but also functionally into typical intestinal epithelial cells under the inductive influence of the duodenal mesenchyme.

Heterotypic induction of stomach-like glands in the urogenital sinus endoderm under the influence of stomach mesenchyme of foetal rats

SummaryUrogenital sinus endoderm of 16.5-day rat foetuses was combined with stomach mesenchyme and the recombinants were either treated with testosterone and grown in vitro or cultured beneath the kidney capsule of adult male rats of the same strain. It was found that testosterone stimulated mitosis in the urogenital endoderm. In recombinants grown under the kidney capsule a stratified squamous epithelium and stomach-like glands were induced under the influence of the forestomach and glandular stomach mesenchymes. However, the induced glands expressed neither rat pepsinogen nor rat ventral prostatic antigen. They did not produce mRNA for the prostatic steroid-binding protein C1. Thus, stomach mesenchyme of rat foetuses induces organ-specific morphogenesis but not functional differentiation in the heterologous endoderm, indicating that cytodifferentiation does not always accompany morphogenesis.

Analysis of epithelial protein profiles of prostatic glands induced heterotypically in the bladder epithelium of the rat

When urinary bladder epithelia of rats were grown in association with fetal urogenital sinus mesenchyme, prostatic morphogenesis was induced. The epithelial proteins were examined by HPLC fractionation followed by sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis (PAGE). More than 500 bands of silver‐stained epithelial proteins were analyzed. The glandular epithelia induced from both adult and fetal bladder epithelia lost all of the 7 bladder‐specific bands (BE 1–7) in most recombinants and expressed a number of prostate‐specific bands. Among the 18 bands commonly found in all prostatic lobes, 13 (PE 4, 7–18) were constantly and 3 (PE 1–3) were sporadically detected, while the other 2 (PE 5 and 6) bands were not detected when the adult epithelium was used in recombination. Among the 7 prostatic lobe‐specific bands (vPE 14, dPE 1–3), most of them were detected when the fetal epithelium was used, while few of them when the adult epithelium was used. These results demonstrate that prostatic morphogenesis induced in the bladder epithelium was associated with most of biochemical features of prostate. In addition to the biochemical study, histological examination revealed that the prostatic differentiation was more complete in the fetal bladder epithelium than the adult one.