Gen Niimi

Embryonic erythropoiesis in human yolk sac: two different compartments for two different processes.

The wall of 12 yolk sacs (YSs) from 17‐ to 50‐day‐old human embryos was examined by light, scanning, and transmission electron microscopy to identify the ontogeny of embryonic erythropoiesis. Initial formation of blood island with the generation of erythroid and endothelial cells was seen in the mesenchymal layer in embryos aged 17 days. A network of blood vessels containing abundant erythroblasts was identified in the YS walls of embryos aged ∼24 days. At this age, erythroblasts were also identified within the embryo body. Primitive erythroblasts were the only cells present within the embryo and its YS until the end of week 5. These cells first appeared in the mesenchymal vascular plexus of the YS wall, and were then observed in the liver and other tissues of the embryo. At embryonic week 5, two compartments were identified in the YS wall; a mesodermal one in which blood vessels were formed, and an endodermal compartment in which erythrocytes were present within the endodermal vesicles. Erythrocytes were small non‐nucleated cells similar to adult erythrocytes. Transmission electron microscopic observation focused on the endodermal vesicles confirmed the presence of definitive erythrocytes only at such extra vascular location. At this age, there were no definitive erythrocytes detected within the embryo. Erythrocytes started to be identified in embryonic blood vessels from week 7 onward. These findings provide information not previously described about YS erythropoiesis during early human development. Microsc. Res. Tech., 2008.

A light and electron microscopic study of the mouse visceral yolk sac endodermal cells in the middle and late embryonic periods, showing the possibility of definitive erythropoiesis

Hematological studies have revealed the importance of the visceral yolk sac (VYS) in the primitive erythropoiesis of mouse embryos at an early stage before day 12. We examined the possibility of the occurrence of extra-embryonic erythropoiesis at a stage later than embryonic day 12 by light and electron microscopic analyses. Surprisingly, a novel structure in the form of erythrocyte-like globules was observed in the VYS endodermal cells. They were consistently present in the VYS endodermal cells from embryonic day 12 until day 18 (birth is day 19), by immunocytochemical and enzyme histochemical analyses. They were immuno-positive for mouse erythrocyte antibody and also positive for the benzidine reaction showing the presence of hemoglobin. The erythrocyte-like globules were shown to be the erythrocytes present in the cytoplasm. These results indicated that erythropoiesis in the VYS endodermal cells continues from the early embryonic stage, as primitive erythropoiesis, until the late stage.

Two different pathways for the transport of primitive and definitive blood cells from the yolk sac to the embryo in humans

During the early human embryonic period nutrients and blood cells are temporarily provided by the extraembryonic yolk sac (YS). The YS before week six is involved not only in primitive but also in definitive erythropoiesis. While the destiny of primitive erythroid cells that fill the blood vessels of the YS is well known, the final destination of erythrocytes present in the endodermal vesicular system is unknown. In the present study we have investigated, step by step, the destiny of the erythrocytes present in the endodermal vesicles during the embryonic period.

Modification of the trypsin cleavage site of rotavirus VP4 to a furin-sensitive form does not enhance replication efficiency

The infectivity of rotavirus (RV) is dependent on an activation process triggered by the proteolytic cleavage of its spike protein VP4. This activation cleavage is performed by exogenous trypsin in the lumen of the intestines in vivo. Here, we report the generation and characterization of a recombinant RV expressing cDNA-derived VP4 with a modified cleavage site (arginine at position 247) recognized by endogenous furin as well as exogenous trypsin. Unexpectedly, the mutant virus (KU//rVP4-R247Furin) was incapable of plaque formation without an exogenous protease, although the mutant VP4s on virions were efficiently cleaved by endogenous furin. Furthermore, KU//rVP4-R247Furin showed impaired infectivity in MA104 and CV-1 cells even in the presence of trypsin compared with the parental virus carrying authentic VP4 (KU//rVP4). Although the total titre of KU//rVP4-R247Furin was comparable to that of KU//rVP4, the extracellular titre of KU//rVP4-R247Furin was markedly lower than its cell-associated titre in comparison with that of KU//rVP4. In contrast, the two viruses showed similar growth in a furin-defective LoVo cell line. These results suggest that intracellular cleavage of VP4 by furin may be disadvantageous for RV infectivity, possibly due to an inefficient virus release process.