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1.
The ascidian egg contains muscle and endoderm determinants that play critical roles in the specification of muscle and endoderm cells, respectively. Endoderm cells of the ascidian embryo express alkaline phosphatase (AP) as a tissue-specific enzyme. We obtained egg fragments from the unfertilized eggs of Ciona savignyi by means of centrifugal force. The largest fragment (red fragments) contained the egg nucleus while other small fragments (black, clear and brown fragments) were anucleate. When inseminated, only red fragments developed into partial embryos, which showed only epidermis cell differentiation and, very rarely, AP activity. When red fragments were fused with other fragments, only black fragments promoted AP expression, suggesting that endoderm determinants were concentrated in the black fragments. A lower dose (1500 J/m2 ) of ultraviolet (UV) light did not eliminate the AP-promoting ability of black fragments, while this dose significantly repressed the ability to promote the expression of the muscle-marker. A higher dose (4500 J/m2 ) of UV light markedly reduced the AP-promoting activity of black fragments. These results suggest that factors for endodermal AP development are inactivated by UV irradiation, but are more resistant than muscle determinants. 相似文献
2.
Mehrdad Salamat Werner Götz Anette Horster Bernd Janotte Rainer Herken 《Cell and tissue research》1993,272(2):375-381
In the present investigation, we examined the role of trophoblast and parietal endoderm cells in the synthesis of carbohydrate-containing components of Reichert's membrane. To eliminate the function of Reichert's membrane as a filter between maternal and embryonal tissues we carried out our examination under in vitro conditions. Parietal yolk sac from mouse embryos on day 9 post coitum (p.c.) were cultivated for 0 to 5 days. Because tannic acid enables a complex formation between carbohydrates and osmium we chose the fixation with this acid for the ultrastructural study. Electron microscopy showed that for assembly of Reichert's membrane, trophoblast cells produce and then release components that were detected as tannic acid-positive granules both in the Reichert's membrane and in the vacuoles of the trophoblast cells. To localize specific carbohydrates we used postembedding-gold-lectin histochemistry on LR-GoldR-embedded tissues. Strong binding sites for the lectins WGA (Triticum vulgare), RCA I (Ricinus communis) and Con A (Canavalia ensiformis) were observed in Reichert's membrane and trophoblast cells but not in the parietal endoderm cells. The LTA (Lotus tetragonolobus)-binding pattern was positive in the membrane and its adjacent cells but that of the LFA (Limax flavus) was negative in the parietal endoderm cells and very weak in Reichert's membrane and trophoblast cells. Our results demonstrate that trophoblast cells are involved in the construction of Reichert's membrane through the production and release of specific glycoconjugates. 相似文献
3.
Angie Rizzino Victor Terranova David Rohrbach Craig Crowley Heather Rizzino 《Journal of cellular biochemistry》1980,13(2):243-253
In this paper we have examined the growth and differentiation of the embryonal carcinoma cell line, F9, in the defined medium EM-3 at low density. We show that the growth of F9 and their differentiated cells (F9-diff) in EM-3 is strongly density dependent. At low cell densities the growth of both cell types is severely limited and most of the cells do not survive. Although this poses a problem for working with F9 and F9-diff in EM-3, it provides a convenient assay for identifying molecules that support their growth at low density. Using this assay, we have determined that laminin, a newly isolated glycoprotein of basement membranes, significantly improves the growth and short-term survival of both F9 and F9-diff. However, addition of laminin to EM-3 is insufficient to promote the clonal growth of these cell types. Our findings also indicate that laminin promotes the attachment of F9 and F9-diff in defined media. On the basis of our results, we propose an attachment function for laminin during the early stages of mammalian development. 相似文献
4.
C. P. Plescia · C. E. Rogler · L. E. Rogler 《Differentiation; research in biological diversity》2001,68(4-5):254-269
5.
6.
Definitive endoderm differentiation is crucial for generating respiratory and
gastrointestinal organs including pancreas and liver. However, whether epigenetic
regulation contributes to this process is unknown. Here, we show that the H3K27me3
demethylases KDM6A and KDM6B play an important role in endoderm differentiation from human
ESCs. Knockdown of KDM6A or KDM6B impairs endoderm differentiation, which can be rescued
by sequential treatment with WNT agonist and antagonist. KDM6A and KDM6B contribute to the
activation of WNT3 and DKK1 at different differentiation stages when WNT3 and DKK1 are
required for mesendoderm and definitive endoderm differentiation, respectively. Our study
not only uncovers an important role of the H3K27me3 demethylases in definitive endoderm
differentiation, but also reveals that they achieve this through modulating the WNT
signaling pathway. 相似文献
7.
Nestor Saiz Minjung Kang Nadine Schrode Xinghua Lou Anna-Katerina Hadjantonakis 《Journal of visualized experiments : JoVE》2016,(108)
This protocol presents a method to perform quantitative, single-cell in situ analyses of protein expression to study lineage specificationin mouse preimplantation embryos. The procedures necessary for embryo collection, immunofluorescence, imaging on a confocal microscope, and image segmentation and analysis are described. This method allows quantitation of the expression of multiple nuclear markers and the spatial (XYZ) coordinates of all cells in the embryo. It takes advantage of MINS, an image segmentation software tool specifically developed for the analysis of confocal images of preimplantation embryos and embryonic stem cell (ESC) colonies. MINS carries out unsupervised nuclear segmentation across the X, Y and Z dimensions, and produces information on cell position in three-dimensional space, as well as nuclear fluorescence levels for all channels with minimal user input. While this protocol has been optimized for the analysis of images of preimplantation stage mouse embryos, it can easily be adapted to the analysis of any other samples exhibiting a good signal-to-noise ratio and where high nuclear density poses a hurdle to image segmentation (e.g., expression analysis of embryonic stem cell (ESC) colonies, differentiating cells in culture, embryos of other species or stages, etc.). 相似文献
8.
Nicolas Fossat Tania Radziewic Vanessa Jones Karin Tourle Patrick P. L. Tam 《Genesis (New York, N.Y. : 2000)》2016,54(3):115-122
Rbm47 encodes a RNA binding protein that is necessary for Cytidine to Uridine RNA editing. Rbm47gt/gt mutant mice that harbor inactivated Rbm47 display poor viability. Here it was determined that the loss of Rbm47gt/gt offspring is due to embryonic lethality at mid‐gestation. It was further showed that growth of the surviving Rbm47gt/gt mutants is impaired. Rbm47 is expressed in both the visceral endoderm and the definitive endoderm. Using the utility of the switchable FlEx gene‐trap cassette and the activity of Cre and FLP recombinases to generate mice that conditionally inactivate and restore Rbm47 function in tissue‐specific manner, it was demonstrated that Rbm47 function is required in the embryo proper, and not the visceral endoderm, for viability and growth. genesis 54:115–122, 2016. © 2016 Wiley Periodicals, Inc. 相似文献
9.
Studies on the genetic basis of rostral-caudal specification, neural induction, and head development require knowledge of the relevant gene expression patterns. Gaps in our understanding of gene expression have led us to examine the detailed spatiotemporal expression patterns of 19 genes implicated in early development, to learn more about their potential role in specifying and patterning early developmental processes leading to head formation. Here, we report the expression patterns of these markers in blastula- and gastrula-stage chick embryos, using whole-mount in situ hybridisation. Nodal, Fgf8, Bmp7, Chordin, Lim1, Hnf3beta, Otx2, Goosecoid, Cerberus, Hex, Dickkopf1, and Crescent are all already expressed by the time the egg is laid. When the primitive streak has reached its full length, a later group of genes, including Ganf, Six3, Bmp2, Bmp4, Noggin, Follistatin, and Qin (BF1), begins to be expressed. We reassess current models of early rostral patterning based on the analysis of these dynamic spatiotemporal expression patterns. 相似文献
10.
Cytokeratins 8 and 19 in the mouse placental development 总被引:9,自引:0,他引:9
Tamai Y Ishikawa T Bösl MR Mori M Nozaki M Baribault H Oshima RG Taketo MM 《The Journal of cell biology》2000,151(3):563-572
To investigate the expression and biological roles of cytokeratin 19 (K19) in development and in adult tissues, we inactivated the mouse K19 gene (Krt1-19) by inserting a bacterial beta-galactosidase gene (lacZ) by homologous recombination in embryonic stem cells, and established germ line mutant mice. Both heterozygous and homozygous mutant mice were viable, fertile, and appeared normal. By 7.5-8.0 days post coitum (dpc), heterozygous mutant embryos expressed lacZ in the notochordal plate and hindgut diverticulum, reflecting the fact that the notochord and the gut endoderm are derived from the axial mesoderm-originated cells. In the adult mutant, lacZ was expressed mainly in epithelial tissues. To investigate the possible functional cooperation and synergy between K19 and K8, we then constructed compound homozygous mutants, whose embryos died approximately 10 dpc. The lethality resulted from defects in the placenta where both K19 and K8 are normally expressed. As early as 9. 5 dpc, the compound mutant placenta had an excessive number of giant trophoblasts, but lacked proper labyrinthine trophoblast or spongiotrophoblast development, which apparently caused flooding of the maternal blood into the embryonic placenta. These results indicate that K19 and K8 cooperate in ensuring the normal development of placental tissues. 相似文献