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N Shiojiri

Publications and source records attributed to N Shiojiri.

At least 19 recordsLinked to original sources
PubMed2001

Immunolocalization of hepatocyte growth factor and its receptor (c-Met) during mouse liver development.

Although hepatocyte growth factor (HGF) was discovered as a potent hepatotrophic factor responsible for liver regeneration and may involve some organ development in embryogenesis, it remains to be revealed what roles HGF plays in liver development. The present study was undertaken to determine which cells express HGF and its receptor c-Met and when c-Met is activated in mouse liver development by using immunoblotting and immunohistochemical techniques. HGF was detected in hepatocytes and non-parenchymal cells, including biliary epithelial cells, periportal connective tissue cells, megakaryocytes, endothelial cells, and sinusoidal cells, throughout liver development. Positive HGF immunostaining in hepatocytes increased during postnatal development, and reached the maximal level in the adult stage. c-Met protein was also expressed in hepatocytes throughout liver development, but maximal staining was obtained in 1- or 2-week-old livers. Phosphorylation of tyrosine residues in the c-Met beta chain also occurred in these stages. These results suggest that HGF signaling is implicated in hepatocyte growth during postnatal liver development, and its action could be in a paracrine mode; HGF produced by non-parenchymal cells such as sinusoidal cells acts on hepatocytes expressing c-Met receptors. Positive immunostaining in adult and postnatal hepatocytes may be derived from their blood clearance of HGF.

Animals↗
PubMed2001

Cell lineage analysis during liver development using the spf(ash)-heterozygous mouse.

Biliary epithelial cells differentiate from periportal hepatoblasts during fetal mouse liver development. It remains to be determined whether each hepatoblast is equivalent for differentiation into hepatocytes and biliary epithelial cells in normal liver development. To resolve this question, the mosaic pattern of ornithine transcarbamylase (OTC) expression was analyzed in the hepatoblast population of spf(ash) (sparse-fur with abnormal skin and hair)-heterozygous fetal mouse livers, in which random inactivation of either the X chromosome carrying the spf(ash) gene (causing OTC deficiency) or its wild-type gene occurs. Aggregates (patches) of OTC-positive hepatoblasts showed very complex patterns, and their shapes and size distributions were similar in sections from periportal regions and nonperiportal regions of the fetal liver in which bile duct differentiation by periportal hepatoblasts occurred. Average sizes of periportal patches were larger than those of nonperiportal patches because of the presence of more hemopoietic cells in the latter region. The OTC mosaicism in periportal bile duct progenitors and hepatoblast islands of other liver parenchyma was also similar. These results suggest that the growth patterns of hepatoblasts are similar in both periportal and nonperiportal regions. Isolated three-dimensional patches comprising hepatoblasts giving rise to only biliary epithelial cells or hepatoblasts giving rise to both hepatocytes and biliary epithelial cells were observed in periportal regions. In nonperiportal regions, patches consisting of hepatoblasts differentiating into hepatocytes were also seen. Thus, it is likely that there are three lineages for the developmental fates of hepatoblasts: hepatoblasts giving rise to only biliary epithelial cells, hepatoblasts giving rise to only hepatocytes, and hepatoblasts giving rise to both of them.

Animals↗
PubMed2000

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.

Aging↗
PubMed2000

Prevention of hepatic apoptosis and embryonic lethality in RelA/TNFR-1 double knockout mice.

Mice deficient in the nuclear factor kappaB (NF-kappaB)-transactivating gene RelA (p65) die at embryonic days 14-15 with massive liver apoptosis. In the adult liver, activation of the NF-kappaB heterodimer RelA/p50 can cause hepatocyte proliferation, apoptosis, or the induction of acute-phase response genes. We examined, during wild-type fetal liver development, the expression of the Rel family member proteins, as well as other proteins known to be important for NF-kappaB activation. We found these proteins and active NF-kappaB complexes in the developing liver from at least 2 days before the onset of lethality observed in RelA knockouts. This suggests that the timing of NF-kappaB activation is not related to the timing of lethality. We therefore hypothesized that, in the absence of RelA, embryos were sensitized to tumor necrosis factor (TNF) receptor 1 (TNFR-1)-mediated apoptosis. Thus, we generated mice that were deficient in both RelA and TNFR-1 to determine whether apoptotic signaling through TNFR-1 was responsible for the lethal phenotype. RelA/TNFR-1 double knockout mice survived embryonic development and were born with normal livers without evidence of increased hepatocyte apoptosis. These animals became runted shortly after birth and survived an average of 10 days, dying from acute hepatitis with an extensive hepatic infiltration of immature neutrophils. We conclude that neither RelA nor TNFR-1 is required for liver development and that RelA protects the embryonic liver from TNFR-1-mediated apoptotic signals. However, the absence of both TNFR-1 signaling and RelA activity in newborn mice makes these animals susceptible to endogenous hepatic infection.

Animals↗
PubMed2000

Immunohistochemical analysis of development of desmin-positive hepatic stellate cells in mouse liver.

Development of desmin-positive hepatic stellate cells was studied in mice using double immunofluorescent techniques and in vitro cultures with special attention given to their cell lineages. Several studies recently reported on the presence of cells that are immunologically reactive with both antidesmin and anticytokeratin antibodies in young fetal rat livers, and suggested the possibility that these cells give rise to hepatocytes and hepatic stellate cells. At early stages of mouse liver development, stellate cells with desmin-positive filaments were scattered in the liver parenchyma. However, the stellate cells definitely differed from hepatoblasts and hepatocytes in terms of their morphology and expression of desmin and hepatoblast and hepatocyte-specific E-cadherin in the liver. Fetal hepatoblasts and hepatocytes did not react with antidesmin antibodies, nor did desmin-positive stellate cells express E-cadherin in vivo and in vitro. Thus it is likely that desmin-positive stellate cells and hepatoblasts belong to different cell lineages. In the fetal liver, the desmin-positive stellate cells surrounded blood vessels, and extended their processes to haematopoietic cells and megakaryocytes. Many, but not all, hepatoblasts and hepatocytes were observed to be associated with the stellate cells. At fetal stages, cellular processes positive for desmin in the stellate cells were also thick compared with those in the adult liver, in which desmin-positive stellate cells lay in Disse's space and were closely associated with all hepatocytes. These developmental changes in the geography of desmin-positive cells in the liver parenchyma and their morphology may be associated with their maturation and interactions with other cell types.

Albumins↗
PubMed1999

Chemical analysis of melanin pigments in feather germs of Japanese quail Bh (black at hatch) mutants.

Bh (black at hatch) is a mutation of Japanese quails which causes darkening or lightening of the plumage in heterozygotes or homozygotes, respectively. We chemically analyzed melanin pigments in feather germs of Bh mutant embryos and in feathers of adult animals. Dark brown dorsal feathers of wild-type adult animals had white barrings, but heterozygous ones lacked clear barrings. The feathers of wild-type and heterozygote animals contained both eumelanins and pheomelanins, the latter being more pheomelanic. On the dorsal skin of 10-day old wild-type embryos, longitudinal stripes from black and yellow rows of feather germs developed; two or three longitudinal rows of black feather germs and then two or three rows of yellow feather germs next to the short central feather germs. Heterozygous embryos appeared black in plumage pigmentation, due to the presence of 'gray' feather germs in rows of dorsal feather germs that corresponded to yellow rows in wild-type embryos. Homozygous dorsal feather germs did not develop the black and yellow longitudinal stripes, but were brown. Chemical analysis showed that embryos of each genotype contained both eumelanins and pheomelanins in the feather germs; however, the eumelanin content in homozygous feather germs was very low. These results suggest that the Bh mutation causes pheomelanic changes in feathers of quails.

Animals↗
PubMed1998

Heterogeneous carbamoylphosphate synthetase I expression in testicular transplants of fetal mouse liver.

Expression of carbamoylphosphate synthetase I (CPSI; EC 6.3.4.16) was examined immunohistochemically in normal development of the mouse liver, and in testicular transplants of fetal liver fragments. CPSI started to be expressed in all hepatocytes around 15 days of gestation, and became heterogeneous (i.e. absent from pericentral hepatocytes) around 2 weeks after birth. Most hepatocytes in fetal liver fragments placed for 2 months under the testicular capsule expressed this enzyme except for the pericentral ones, most of which were positively stained with anti-glutamine synthetase (GS; EC 6.3.1.2) antiserum. This distribution resembled that in the adult liver. The steep change in CPSI immunostaining in liver lobules suggests that the microenvironment tightly connected to the central veins plays an important role in the suppression of CPSI expression in the pericentral hepatocytes. Some pericentral hepatocytes were also negative for both enzymes. Thus, control mechanisms of CPSI expression may be different from those of GS expression in pericentral hepatocytes.

Animals↗
PubMed1997

Development and differentiation of bile ducts in the mammalian liver.

The development and differentiation of bile ducts in the human and rodent liver are reviewed. The liver primordium develops as a ventral diverticulum in the anterior intestinal portal region, which consists of endodermal and mesodermal components. The endodermal cells differentiate into hepatocytes and all epithelial cells of the bile ducts in the adult liver. The gallbladder and extrahepatic bile ducts also start to develop from hepatic endodermal cells and hepatoblasts just after liver primordium formation. The gallbladder and cystic duct do not develop through hepatic development in the rat. Intrahepatic bile ducts are formed from periportal hepatoblasts forming the "ductal plate" and expressing alpha-fetoprotein, and albumin and bile duct-specific cytokeratin and develop independently of extrahepatic bile duct formation. The first sign of intrahepatic bile duct differentiation is the increased expression of bile duct-specific cytokeratin and large lumina formation in periportal hepatoblasts, and then deposition of basal laminar components occurs on the basal side. Their development takes place discontinuously along portal veins at the early stage of development, and they then become confluent through development. Periportal connective tissue, glucocorticoid hormones, and basal laminar components may play important roles in the differentiation of bile ducts.

Albumins↗
PubMed1997

Complementary expression of glutamine synthetase and carbamoylphosphate synthetase I in ornithine carbamoyltransferase-deficient mouse liver (spf-ash mouse).

Glutamine synthetase and carbamoylphosphate synthetase I expression was examined immunohistochemically in livers of spf-ash homozygous and hemizygous mice, in which one of the urea cycle enzymes (ornithine carbamoyltransferase) is deficient and hyperammonemic disorders are obvious. In the mutant adult mouse liver, only hepatocytes lining central veins expressed glutamine synthetase. In contrast, other hepatocytes expressed carbamoylphosphate synthetase I but not glutamine synthetase. This complementary expression pattern is similar to that seen in wild-type mouse liver. In the liver of mutant young mice, which showed severe retarded growth and abnormal hair and skin development, the developmental expression pattern of both enzymes was also similar to that of the corresponding wild-type liver. However, suppression of carbamoylphosphate synthetase I expression in the pericentral hepatocytes occurred later in the mutant than in wild-type liver. These results show that high plasma concentrations of ammonium ions, which are one of the substrates for both the enzymes, do not change their complementary expression. Instead they support the idea that factor(s) associated with central veins rather than humoral factors direct pericentral hepatocytes to express glutamine synthetase and to suppress carbamoylphosphate synthetase I expression.

Ammonia↗
PubMed1997

Bh (black at hatch) gene appears to be expressed in melanocytes of feather germs in the Japanese quail (Coturnix coturnix japonica).

The Bh (black at hatch) gene was examined to determine whether it is expressed in plumage melanocytes by analyzing pigmentation patterns of Bh melanocytes placed in the micro-environment of the feather germs of quail embryos with pink eyes. These host quails genetically lack a large part of plumage melanin. The Bh locus in these almost white quails is wild-type. When Bh neural crest cells were transplanted orthotopically into the host embryos, wild-type and Bh/+ melanocytes, which differentiated from the transplanted neural crest cells, formed plumage pigmentation patterns characteristic of each genotype in the micro-environment of the host feather germs. Brown plumage pigmentation, which was very similar to that of 10-day Bh/Bh embryos, was also observed in the feather germs of host embryos that received Bh neural crest cells, although the genotype of the donors could not be determined. These donors died before pigmentation of their feather germs occurred. The results demonstrate that pigmentation patterns of Bh melanocytes are not altered in the micro-environment of the host germs, suggesting that the Bh gene is autonomous in Bh melanocytes and is expressed in melanocytes of both Bh and the host feather germs, and that it causes the normal pigmentation pattern to be altered.

Animals↗
PubMed1997

Differentiation of biliary epithelial cells from the mouse hepatic endodermal cells cultured in vitro.

Differentiation of biliary epithelial cells from hepatic endodermal cells of the mouse embryo was examined with a special attention to the role of the connective tissue. When the whole liver primordium of the 9.5-day mouse embryo was cultured in vitro for 5 days, the endodermal cells differentiated into mature hepatocytes expressing carbamoylphosphate synthetase I (CPSI) and accumulating glycogen. Intrahepatic bile duct cells and connective tissue were poorly developed in this culture. However, when the hepatic endoderm was recombined with the 4-day embryonic chick lung mesenchyme and cultured in vitro, the endodermal cells differentiated into many ductal epithelial cells as well as mature hepatocytes with abundant connective tissue development. These results suggest that the ducts might be bile ducts, and that connective tissue is very important for bile duct development. In addition, this in vitro culture system might be useful for the study of mechanisms of bile duct differentiation and congenital biliary atresia.

Animals↗
PubMed1997

Mosaic pattern of ornithine transcarbamylase expression in spfash mouse liver.

Mosaicism of ornithine transcarbamylase expression was immunohistochemically examined in hepatocytes of spfash heterozygous female mouse livers. An immunohistochemical method using polyclonal antibodies against ornithine transcarbamylase visualized the mosaicism in serial paraffin sections. Very complicated mosaic patterns consisting of positive and negative hepatocytes were obtained in a section of adult liver, but a computer-aided three-dimensional analysis of serial sections demonstrated that patches with complicated shapes and various sizes, which are contiguous groups of positive or negative hepatocytes and are isolated in sections, connected very well with one another. No definite orientation such as portal-central was observed in the three-dimensional images of each patch. In balanced regions of the mosaicism, most individual plates along their straight portal-central lengths did not appear to be composed of only marker-positive or marker-negative hepatocytes. By contrast, in unbalanced regions of the mosaicism, individual plates along their total portal-central lengths often consisted entirely of a major type of hepatocytes. A patch appeared to be present in more than two lobules, but each patch did not constitute a complete lobule. Complicated mosaic patterns of patches were also seen in neonatal and postnatal livers. These results suggest that, although hepatocytes proliferate and migrate extensively during development, they might allocate their daughter cells contiguously and the orientation of their allocation might be random, leading to the formation of three-dimensionally large contiguous quasiclones of hepatocytes, the shapes of which are very complicated.

Animals↗
PubMed1996

Differentiation of the mouse hepatic primordium cultured in vitro.

The differentiation of hepatic endodermal cells is affected by endodermal-mesodermal interactions. To examine the control mechanisms of this differentiation, we cultured mouse liver primordium and tissue recombinants of the hepatic endoderm with homo- or heterologous mesenchyme in vitro. When the hepatic primordia at somite stages 15-23 were cultured in vitro for 5-10 days, the endodermal cells differentiated into large hepatocytes expressing alpha-fetoprotein (AFP), albumin and carbamoylphosphate synthetase I (CPSI) and storing glycogen. AFP continued to be expressed in hepatocytes through culture for 10 days. Albumin and CPSI expression started in hepatocytes at 1 and 2 days after culture, respectively. Dexamethasone stimulated hepatocyte differentiation (expression of CPSI and glycogen accumulation) and large lumen formation of hepatocytes, but it did not change the commencement of differentiation. When the hepatic endoderm was recombined with hepatic mesenchyme or 4-day embryonic chick lung mesenchyme, clotted in Matrigel, which is a basement-membrane-like substratum, and cultured for 5 days in vitro, it differentiated into large hepatocytes expressing albumin and CPSI and accumulating glycogen. Lung mesenchyme promoted duct formation more efficiently than the hepatic mesenchyme did. However, the hepatic endodermal cells failed to differentiate into large hepatocytes when cultured with 6-day embryonic chick metanephric mesenchyme or with 2.5-day chick somitic mesenchyme, or cultured alone in Matrigel, suggesting that the endodermal cells require the presence of splanchnic mesoderm for their differentiation in vitro. Addition of HGF (hepatocyte growth factor), aFGF (acidic fibroblast growth factor), or bFGF (basic fibroblast growth factor) also did not support the survival of hepatic endodermal cells or hepatocyte differentiation in culture without mesenchyme. Matrigel and those growth factors might not be a suitable substitute for the mesenchyme.

Albumins↗
PubMed1995

Bh (black at hatch) gene appears to cause whole-body hemorrhage in homozygous embryos of Japanese quail (Coturnix coturnix japonica).

Genotypes of early embryos obtained from mating Bh/+ quail were determined by chorio-allantoic membrane grafting of their thigh skin, and the development of the Bh homozygous embryos was examined histologically. The Bh homozygous embryos died at an early stage of development (after 7 to 10 days of incubation) with a very high incidence of whole-body hemorrhage (approximately 100%), but malformation of neural tubes was not always accompanied by Bh homozygosity. Degeneration of the liver was more frequently observed in homozygous embryos than in wild-type and heterozygous embryos. The results suggest that the Bh gene causes whole-body hemorrhage and subsequent death of homozygous embryos. In addition, our data indicate that Bh homozygotes would develop brown feathers on their skin if they were able to survive.

Allantois↗
PubMed1995

Heterogeneous hepatocellular expression of glutamine synthetase in developing mouse liver and in testicular transplants of fetal liver.

BACKGROUND: Glutamine synthetase is exclusively expressed in pericentral hepatocytes in mammalian liver, but its regulation mechanism is still largely unknown. EXPERIMENTAL DESIGN: Heterogeneous expression of glutamine synthetase was examined in detail during mouse liver development by immunohistochemistry and by in situ hybridization. Heterogeneous expression of this enzyme was also analyzed in immature liver fragments transplanted to an ectopic site where no portal blood flow exists. RESULTS: At 18.5 days of gestation, a random, spotty distribution of low levels of glutamine synthetase mRNA was observed all over the liver parenchyma, but the enzyme protein was not detectable immunohistochemically in the liver at any fetal stage. Glutamine synthetase and its mRNA began to be heterogeneously expressed in pericentral hepatocytes 2 to 3 days after birth, when glycogen accumulation in the liver parenchyma was rather homogeneous. In the early postnatal development, a mosaic distribution of positive and negative hepatocytes with respect to glutamine synthetase protein and mRNA was noted around the central veins. Subsequently, mRNA distribution gradually became continuous, although some hepatocytes still lacked protein, indicating partial regulation at the translational level. When fetal liver fragments that had not yet heterogeneously expressed glutamine synthetase were cultured under the testis capsule of male mice, only pericentral hepatocytes expressed this enzyme after 2 months. However, the distribution of glutamine synthetase protein- and mRNA-positive hepatocytes around the central veins was patchy rather than continuous, as in perinatal livers. CONCLUSIONS: These results support the importance of local interactions of hepatocytes with intrahepatic cell populations and/or structural elements. Furthermore, they demonstrate that the capacity for the positional expression of glutamine synthetase is already established at a fetal age before expression of glutamine synthetase can be detected.

Animals↗
PubMed1994

Transient expression of bile-duct-specific cytokeratin in fetal mouse hepatocytes.

The differentiation of hepatocytes and biliary epithelial cells has been histochemically analyzed with anti-calf cytokeratin antiserum in the fetal mouse liver. Almost all young fetal hepatocytes transiently express bile-duct-specific cytokeratin; subsequently, the strong staining of the cytokeratin is confined to progenitor cells of intrahepatic biliary epithelial cells around portal veins. These results suggest that all fetal hepatocytes are bi-potent in terms of the differentiation of mature hepatocytes and intrahepatic bile-duct cells, and that the microenvironment around portal veins plays an important role in bile-duct differentiation. Large periportal hepatocytes continue to stain weakly for cytokeratin until 2 weeks after birth, although the number of positive hepatocytes decreases with development. The differentiation of bile ducts from periportal hepatocytes may continue for 2 weeks after birth.

Albumins↗
PubMed1993

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.

Animals↗