Hubrecht Institute Centennial - From embryos to stem cells.

During the last 100 years, the Hubrecht Institute evolved from a small laboratory aimed at providing research material to the scientific community to a modern, fully equipped research institute with state-of-the-art infrastructure, performing research at the highest standard. The past 100 years have been eventful for the Hubrecht Institute with many glorious moments, but also threats to be shut down on several occasions. Here, we will briefly review the rich history of the Hubrecht Institute.

Phosphatidylinositol transfer protein in murine embryonal carcinoma cells during retinoic acid-induced differentiation.

Phosphatidylinositol transfer protein (PI-TP) was studied in P19 embryonal carcinoma (EC) cells at different stages of retinoic acid (RA) induced differentiation. Western blot analysis indicated an increased expression of PI-TP (35 kDa) during differentiation. Western blots of isoelectric focusing gels showed that the 35 kDa band consisted of the PI-carrying form of PI-TP (pl 5.5) and of a novel, more acidic form of PI-TP (pl 5.4), levels of both of which increased during differentiation. These increased levels were not reflected in the in vitro PI-transfer activity of the cytosolic fraction nor in the mRNA levels as analyzed by northern blotting. By using indirect immunofluorescence it was shown that PI-TP is localized in the cytoplasm and associated with perinuclear Golgi structures and that this distribution is slightly affected during RA-induced differentiation. Immunoprecipitation of PI-TP from [32 P]Pi labeled cells demonstrated that the level of phosphorylation of PI-TP is high in undifferentiated P19 EC cells and low after 5 days of RA-induced differentiation. These results strongly suggest that changes in the levels of PI-TP are intimately connected with changes in the growth characteristics of P19 EC cells during RA-induced differentiation. It remains to be established to what extent this connection is governed by the recent finding that PI-TP is an essential cytosolic factor in stimulating phospholipase C activity.

Dye-coupling between blastomeres in early embryos ofPatella vulgata (mollusca, gastropoda): Its relevance for cell determination.

During the early development of the molluscPatella, the dorsoventral axis is established after the fifth cleavage due to direct interaction between the animal micromeres and one of the vegetal macromeres. This vegetal macromere is thereby induced to become the mesentoblast mother cell (3D). In this study we have examined intercellular communication in earlyPatella embryos by monitoring the transfer of the fluorescent dye, Lucifer Yellow, upon iontophoretic injection into blastomeres between the second and sixth cleavage. Up to the fifth cleavage dye transfer is detectable neither inin toto embryos nor in serial sections. Shortly after the fifth cleavage dye-coupling between blastomeres becomes apparent. This occurs approximately 40 min before the interaction between animal micromeres and the future mesentoblast mother cell. Inspection of serially sectioned embryos after dye-iontophoresis in either animal micromeres or in the central macromere 3D showed the absence of direct dye-transfer between these cells at the stage of interaction. The reduced rate of dye-transfer from the 3D macromere to its dorsal neighbour 2d2 suggests a bilateral symmetrical transfer pattern, the axis of which corresponds with the dorsoventral axis at the sixth cleavage. Cell deletion experiments demonstrated that the establishment of dye-coupling between the vegetal macromeres occurs independently of the interaction between animal and vegetal blastomeres.