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11.
The initial period of mammalian embryonic development is primarily devoted to cell commitment to the pluripotent lineage, as well as to the formation of extraembryonic tissues essential for embryo survival in utero. This phase of development is also characterized by extensive morphological transitions. Cells within the preimplantation embryo exhibit extraordinary cell plasticity and adaptation in response to experimental manipulation, highlighting the use of a regulative developmental strategy rather than a predetermined one resulting from the non-uniform distribution of maternal information in the cytoplasm. Consequently, early mammalian development represents a useful model to study how the three primary cell lineages; the epiblast, primitive endoderm (also referred to as the hypoblast) and trophoblast, emerge from a totipotent single cell, the zygote. In this review, we will discuss how the isolation and genetic manipulation of murine stem cells representing each of these three lineages has contributed to our understanding of the molecular basis of early developmental events. 相似文献
12.
eXtraembryonic ENdoderm (XEN) stem cells produce factors that activate heart formation 总被引:1,自引:0,他引:1
Background
Initial specification of cardiomyocytes in the mouse results from interactions between the extraembryonic anterior visceral endoderm (AVE) and the nascent mesoderm. However the mechanism by which AVE activates cardiogenesis is not well understood, and the identity of specific cardiogenic factors in the endoderm remains elusive. Most mammalian studies of the cardiogenic potential of the endoderm have relied on the use of cell lines that are similar to the heart-inducing AVE. These include the embryonal-carcinoma-derived cell lines, END2 and PYS2. The recent development of protocols to isolate eXtraembryonic ENdoderm (XEN) stem cells, representing the extraembryonic endoderm lineage, from blastocyst stage mouse embryos offers new tools for the genetic dissection of cardiogenesis.Methodology/Principal Findings
Here, we demonstrate that XEN cell-conditioned media (CM) enhances cardiogenesis during Embryoid Body (EB) differentiation of mouse embryonic stem (ES) cells in a manner comparable to PYS2-CM and END2-CM. Addition of CM from each of these three cell lines enhanced the percentage of EBs that formed beating areas, but ultimately, only XEN-CM and PYS2-CM increased the total number of cardiomyocytes that formed. Furthermore, our observations revealed that both contact-independent and contact-dependent factors are required to mediate the full cardiogenic potential of the endoderm. Finally, we used gene array comparison to identify factors in these cell lines that could mediate their cardiogenic potential.Conclusions/Significance
These studies represent the first step in the use of XEN cells as a molecular genetic tool to study cardiomyocyte differentiation. Not only are XEN cells functionally similar to the heart-inducing AVE, but also can be used for the genetic dissection of the cardiogenic potential of AVE, since they can be isolated from both wild type and mutant blastocysts. These studies further demonstrate the importance of both contact-dependent and contact-independent factors in cardiogenesis and identify potential heart-inducing proteins in the endoderm. 相似文献13.
Properties of Leaf NAD-Malic Enzyme from the Inducible Crassulacean Acid Metabolism Species Mesembryanthemum crystallinum 总被引:1,自引:0,他引:1
NAD-malic enzyme (NAD-ME) functions to decarboxylate malatein the light in leaves of certain species displaying Crassulaceanacid metabolism (CAM). The properties of NAD-ME in desaltedextracts from the inducible CAM species, Mesembryanthemum crystallinumwere examined. The shapes of the malate saturation curve andthe activity versus pH curve at 10 mM malate were dependenton the presence of the activator CoA. The malate saturationcurve was sigmoidal in the absence of an activator and hyperbolicin the presence of CoA. The pH optimum with 10mM malate andMn2+ as cofactor was as low as 6.5 without an activator, andincreased to 7.2 in the presence of CoA. Fumarate activationwas synergistic with CoA above pH 7.2. The enzyme displayedhysteretic behavior under suboptimal assay conditions. Rapid extraction and desalting of the enzyme (<1.5 mim) followedimmediately by assay did not reveal any difference in the propertiesof the enzyme on a day/night basis. It is proposed that diurnalregulation of the enzyme in vivo is mediated by pH and malatelevel without a change in the oligomeric form of the enzyme.The molecular weight of the enzyme was approximately 350,000at pH 6.5 or 7.8. The enzyme obtained from M. crystallinum inthe C3 mode was very similar to the CAM enzyme except that itdisplayed a lower Vmax.
3 Current address: MSU-DOE Plant Research Lab, Michigan StateUniversity, E. Lansing, Michigan, U.S.A. 48824. (Received October 2, 1984; Accepted December 20, 1984) 相似文献
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15.
Kemar Brown Stephanie Legros Jér?me Artus Michael Xavier Doss Raya Khanin Anna-Katerina Hadjantonakis Ann Foley 《PloS one》2010,5(8)
Prior to gastrulation in the mouse, all endodermal cells arise from the primitive
endoderm of the blastocyst stage embryo. Primitive endoderm and its derivatives
are generally referred to as extra-embryonic endoderm (ExEn) because the
majority of these cells contribute to extra-embryonic lineages encompassing the
visceral endoderm (VE) and the parietal endoderm (PE). During gastrulation, the
definitive endoderm (DE) forms by ingression of cells from the epiblast. The DE
comprises most of the cells of the gut and its accessory organs. Despite their
different origins and fates, there is a surprising amount of overlap in marker
expression between the ExEn and DE, making it difficult to distinguish between
these cell types by marker analysis. This is significant for two main reasons.
First, because endodermal organs, such as the liver and pancreas, play important
physiological roles in adult animals, much experimental effort has been directed
in recent years toward the establishment of protocols for the efficient
derivation of endodermal cell types in vitro. Conversely,
factors secreted by the VE play pivotal roles that cannot be attributed to the
DE in early axis formation, heart formation and the patterning of the anterior
nervous system. Thus, efforts in both of these areas have been hampered by a
lack of markers that clearly distinguish between ExEn and DE. To further
understand the ExEn we have undertaken a comparative analysis of three ExEn-like
cell lines (END2, PYS2 and XEN). PYS2 cells are derived from embryonal
carcinomas (EC) of 129 strain mice and have been characterized as parietal
endoderm-like [1], END2 cells are derived from P19 ECs and
described as visceral endoderm-like, while XEN cells are derived from blastocyst
stage embryos and are described as primitive endoderm-like. Our analysis
suggests that none of these cell lines represent a bona fide
single in vivo lineage. Both PYS2 and XEN cells represent mixed
populations expressing markers for several ExEn lineages. Conversely END2 cells,
which were previously characterized as VE-like, fail to express many markers
that are widely expressed in the VE, but instead express markers for only a
subset of the VE, the anterior visceral endoderm. In addition END2 cells also
express markers for the PE. We extended these observations with microarray
analysis which was used to probe and refine previously published data sets of
genes proposed to distinguish between DE and VE. Finally, genome-wide pathway
analysis revealed that SMAD-independent TGFbeta signaling through a TAK1/p38/JNK
or TAK1/NLK pathway may represent one mode of intracellular signaling shared by
all three of these lines, and suggests that factors downstream of these pathways
may mediate some functions of the ExEn. These studies represent the first step
in the development of XEN cells as a powerful molecular genetic tool to study
the endodermal signals that mediate the important developmental functions of the
extra-embryonic endoderm. Our data refine our current knowledge of markers that
distinguish various subtypes of endoderm. In addition, pathway analysis suggests
that the ExEn may mediate some of its functions through a non-classical MAP
Kinase signaling pathway downstream of TAK1. 相似文献
16.
Plusa B Piliszek A Frankenberg S Artus J Hadjantonakis AK 《Development (Cambridge, England)》2008,135(18):3081-3091
The first two lineages to differentiate from a pluripotent cell population during mammalian development are the extraembryonic trophectoderm (TE) and the primitive endoderm (PrE). Whereas the mechanisms of TE specification have been extensively studied, segregation of PrE and the pluripotent epiblast (EPI) has received comparatively little attention. A current model of PrE specification suggests PrE precursors exhibit an apparently random distribution within the inner cell mass of the early blastocyst and then segregate to their final position lining the cavity by the late blastocyst. We have identified platelet-derived growth factor receptor alpha (Pdgfralpha) as an early-expressed protein that is also a marker of the later PrE lineage. By combining live imaging of embryos expressing a histone H2B-GFP fusion protein reporter under the control of Pdgfra regulatory elements with the analysis of lineage-specific markers, we investigated the events leading to PrE and EPI lineage segregation in the mouse, and correlated our findings using an embryo staging system based on total cell number. Before blastocyst formation, lineage-specific factors are expressed in an overlapping manner. Subsequently, a gradual progression towards a mutually exclusive expression of PrE- and EPI-specific markers occurs. Finally, cell sorting is achieved by a variety of cell behaviours and by selective apoptosis. 相似文献
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18.
Nancy N. Artus Shauna C. Somerville Christopher R. Somerville George H. Lorimer 《植物科学评论》1986,4(2):121-147
Since the discovery of RuBP oxygenase activity more than a decade ago, our understanding of the sequence of metabolic events associated with photorespiratory activity in C3 plants has matured considerably. A coherent model of photorespiratory metabolism has been substantiated by a wide variety of experimental approaches and most photorespiratory phenomena can be satisfactorily explained. By contrast, the issues pertaining to the regulation of photorespiratory activity by genetic or chemical means have not been resolved. In addition, the wealth of biochemical, physiological, and genetic information available about photorespiratory metabolism offers as yet unexploited opportunities to investigate the complex cellular processes which organize and coordinate a series of reactions in three distinct metabolic compartments in leaf cells. 相似文献
19.
BMP4 signaling directs primitive endoderm-derived XEN cells to an extraembryonic visceral endoderm identity 总被引:1,自引:0,他引:1
Artus J Douvaras P Piliszek A Isern J Baron MH Hadjantonakis AK 《Developmental biology》2012,361(2):245-262
The visceral endoderm (VE) is an epithelial tissue in the early postimplantation mouse embryo that encapsulates the pluripotent epiblast distally and the extraembryonic ectoderm proximally. In addition to facilitating nutrient exchange before the establishment of a circulation, the VE is critical for patterning the epiblast. Since VE is derived from the primitive endoderm (PrE) of the blastocyst, and PrE-derived eXtraembryonic ENdoderm (XEN) cells can be propagated in vitro, XEN cells should provide an important tool for identifying factors that direct VE differentiation. In this study, we demonstrated that BMP4 signaling induces the formation of a polarized epithelium in XEN cells. This morphological transition was reversible, and was associated with the acquisition of a molecular signature comparable to extraembryonic (ex) VE. Resembling exVE which will form the endoderm of the visceral yolk sac, BMP4-treated XEN cells regulated hematopoiesis by stimulating the expansion of primitive erythroid progenitors. We also observed that LIF exerted an antagonistic effect on BMP4-induced XEN cell differentiation, thereby impacting the extrinsic conditions used for the isolation and maintenance of XEN cells in an undifferentiated state. Taken together, our data suggest that XEN cells can be differentiated towards an exVE identity upon BMP4 stimulation and therefore represent a valuable tool for investigating PrE lineage differentiation. 相似文献
20.
L. Sibille S. Guillemard M.-C. Eberle-Pouzeratte S. Thezenas C. Espitalier-Riviere P. Faurous P.-J. Lamy M. Rossi J.-C. Artus 《Médecine Nucléaire》2010,34(2):78-87
Aim18F-FDG PET/CT by combining both metabolic and anatomical informations has proven to be an effective modality for detecting many types of cancer. Some differentiated forms of cancer like differentiated thyroid carcinoma (DTC) are less FDG avid and thus less easily detectable. Nevertheless 18F-FDG PET/CT has been proved useful in DTC especially in case of suspected recurrent disease with negative whole-body radioiodine scintigraphy (131I WBS) and elevated thyroglobulin (Tg) or thyroglobulin autoantibodies (AbTg) levels. Impact on clinical management after 18F-FDG PET/CT examinations has been analyzed in patients with suspected recurrent DTC in this retrospective study.MethodologyFifty-five 18F-FDG PET/CT were performed in 45 patients with suspected recurrent or residual disease either because of elevated Tg/AbTg levels (n = 45) or uncertain conventional imaging (n = 10) including 131I WBS, cervical echography and CT scan if necessary. 18F-FDG PET/CT results were compared with histopatology and/or clinical follow-up with evaluation of impact on clinical management.ResultsTwenty-nine exams were positive (53 %). There were 20 true-positive (TP) (14 locoregional relapses and six with distant metastases) and nine false-positive (FP) (all cervical). SUVmax median values of hypermetabolic foci were significantly higher in TP (5.1) than in FP (2.8). Overall, 20 (36 %) 18F-FDG PET/CT directly affected clinical management resulting in 13 (65 %) new surgical operations. Sensitivity, specificity, predictive positive value, predictive negative value and accuracy of 18F-FDG PET/CT were estimated for the whole group (respectively 83 %, 71 %, 69 %, 85 % and 76 %) and for two subgroups depending on Tg level (less or more than 1.2 ng/mL).Discussion and conclusion18F-FDG PET/CT is a powerful and useful tool in patients with suspected DTC recurrence or residual disease and should be systematically performed when basal Tg level is above 1.2 ng/mL. Thanks to given anatomical informations it can guide surgical re-operation. 相似文献