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PubMed1995

Strain-dependent epithelial defects in mice lacking the EGF receptor.

Mice and cells lacking the epidermal growth factor receptor (EGFR) were generated to examine its physiological role in vivo. Mutant fetuses are retarded in growth and die at mid-gestation in a 129/Sv genetic background, whereas in a 129/Sv x C57BL/6 cross some survive until birth and even to postnatal day 20 in a 129/Sv x C57BL/6 x MF1 background. Death in utero probably results from a defect in the spongiotrophoblast layer of the placenta. Newborn mutant mice have open eyes, rudimentary whiskers, immature lungs, and defects in the epidermis, correlating with the expression pattern of the EGFR as monitored by beta-galactosidase activity. These defects are probably cell-autonomous because chimeric mice generated with EGFR-/- embryonic stem cells contribute small amounts of mutant cells to some organs. These results indicate that the EGFR regulates epithelial proliferation and differentiation and that the genetic background influences the resulting phenotype.

Animals↗
PubMed1989

Directing cell division during development.

Several evolutionarily conserved proteins constitute a universal mitotic trigger that is precisely controlled during the orderly cell divisions of embryogenesis. As development progresses, the mechanisms controlling this trigger change. Early divisions are executed by maternally synthesized gene products, and in Xenopus they are timed by the accumulation and periodic degradation of cyclin, a trigger component. Later, the zygotic genome assumes control, and in Drosophila, zygotic transcription is required for production of another trigger protein, the product of string. After this transition to zygotic control, pulses of string transcription define the timing of highly patterned embryonic cell divisions and cyclin accumulation is not rate limiting.

Animals↗
PubMed1993

Induction of embryonic major histocompatibility complex antigen expression by gamma-IFN.

Preimplantation mouse embryos were incubated in vitro with mouse recombinant gamma-interferon (IFN). The effect of the gamma-IFN on major histocompatibility complex (MHC) class I antigen expression was tested using an ELISA procedure. It was found that there is a doubling of Db antigens and a tripling of Qa-2 antigens on C57BL/6 mouse embryos cultured from the 8-cell stage for 24 h in the presence of 10(5) units/ml gamma-IFN. The effect of gamma-IFN on the rate of preimplantation embryonic development was tested by culturing 2-cell embryos for 48 h and 8-cell embryos for 24 h in the presence of varying concentrations of gamma-IFN up to 10(6) units/ml. Two methods were used to assess the cell number per embryo after the culture period: incorporation of [3H]thymidine into DNA, and direct counting of nuclei in fixed and stained embryos. Both methods showed that treatment with gamma-IFN increases the rate of development of preimplantation mouse embryos. Since rate of preimplantation embryonic development is genetically controlled by the Ped gene, it is suggested that gamma-IFN has a direct effect on the Ped gene phenotype of preimplantation mouse embryos.

Animals↗
PubMed1999

Stu-7/air-2 is a C. elegans aurora homologue essential for chromosome segregation during embryonic and post-embryonic development.

We have isolated a new sterile uncoordinated C. elegans mutant, stu-7, which is defective in post-embryonic cell divisions in a regionally-specific fashion. The anterior of the worm is relatively unaffected whereas the mid-body and/or posterior are markedly thin, often resulting in worms having a central 'waist'. We have cloned stu-7 and found that it encodes a member of the recently expanding aurora sub-family of serine/threonine kinases. Elimination of maternal as well as zygotic stu-7 expression reveals that stu-7 is essential for mitosis from the first embryonic cell cycle onwards and is required for chromosome segregation though not for centrosome separation or for setting up a bipolar spindle. Multicopy expression of stu-7 also causes mitotic defects, suggesting that the level of this protein must be tightly controlled in order to maintain genetic stability during development.

Amino Acid Sequence↗
PubMed1981

Asymmetric division of lymphoid cells of the thymus of embryonic guinea-pigs. A cell kinetic study.

Cell division of thymus lymphoid cells from 30- to 60-day-old embryonic guinea-pigs, as well as young adults was investigated on cell smears stained with Giemsa. Asymmetrically dividing cells were found in the developmental stage of thymocytes. At the telophase of such cells, the cytoplasma of one of two immature daughter cells was apparently larger in amount than that of the counterpart cell. When 40-day embryonic lymphoid cells were examined, most of the asymmetrically dividing cells at telophase belonged to the larger cell population (mean diameter 15.7 micrometer). The mean diameters of the larger and smaller immature daughter cells of these dividing cells were estimated as 12.0 and 9.2 micrometer, respectively. Ultrastructural study also revealed a lymphoid cell which was under-going in situ apparent symmetric division at telophase. One of two immature daughter cells alone possessed a large cytoplasmic protrusion with few organella. The frequence of asymmetric division calculated by a rough estimation was around 10% of the total cell division between days 30 and 40 of embryonic development, and decreased thereafter. During this developmental period, thymic lymphoid cells were shown to progressively transform into smaller cells. A scheme regarding the mode of thymocyte proliferation during the embryonic period is proposed. and the biological meaning of the present phenomenon is discussed.

Animals↗
PubMed2000

Estrogen receptor-mediated effects of a xenoestrogen, bisphenol A, on preimplantation mouse embryos.

The effects of bisphenol A, a xenoestrogen widely used in industry and dentistry, were studied in early preimplantation mouse embryos. Two-cell mouse embryos were cultured with 100 pM to 100 microM bisphenol A with or without 100 nM tamoxifen and evaluated at 24-h intervals for their development to eight-cell and blastocyst stages. At 72 h, blastocysts were cultured for another 48 h without bisphenol A, and surface areas of trophoblast spread were measured. At 24 h, more embryos exposed to 3 nM bisphenol A than to controls had reached the eight-cell stage. At 48 h, more embryos exposed to 1 nM and 3 nM bisphenol A than to controls had become blastocysts. At 100 microM, bisphenol A decreased frequency of development to blastocysts. Tamoxifen counteracted both stimulatory and inhibitory effects of bisphenol A on blastocyst formation. Although bisphenol A did not alter blastocyst morphology or cell number, early exposure to 100 microM bisphenol A increased subsequent trophoblast areas. These findings suggest that bisphenol A may not only effect early embryonic development via estrogen receptors even at low, environmentally relevant doses, but also exert some late effects on subsequent development of these embryos.

Animals↗
PubMed1984

Effects of alpha-momorcharin on preimplantation development in the mouse.

When alpha- momorcharin was injected intraperitoneally (0.2 mg/25 g body weight) into pregnant mice on Days 1-3 of pregnancy, over 50% of the mice failed to support an implantation. In-vitro study of the effects of the protein on preimplantation embryos showed that the protein did not significantly disturb embryonic development from the 2-cell to compacting morula stage except when high concentrations (greater than or equal to 0.5 microgram/ml) of protein were present. In many embryos, compaction of blastomeres was incomplete and subsequent blastocyst formation was impaired. Other protein-treated embryos that formed compacted morulae and early blastocysts later showed decompaction and degenerated. The protein-treated embryos generally had fewer numbers of cells because cell division beyond the morula stage was impaired. The poor development of morulae may be the cause of inhibition of early pregnancy in the mouse by alpha- momorcharin .

Animals↗
PubMed1996

Effects of cell cycle specific exposure to 3H-thymidine or 3H-arginine on development and cell proliferation of mouse embryos.

One-cell mouse embryos were exposed to either 3H-thymidine (100 or 200 kBq/ml) or 3H-arginine (2.5 to 50 kBq/ml) for 2 h either in G1, S or G2 phase. 3H-Arginine affected embryonic development and cell proliferation in an activity-dependent way irrespective of the cell cycle stage exposed, whereas 3H-thymidine was effective only at higher activities and only after exposure during S phase.

Animals↗
PubMed1999

p27(Kip1) links cell proliferation to morphogenesis in the developing organ of Corti.

Strict control of cellular proliferation is required to shape the complex structures of the developing embryo. The organ of Corti, the auditory neuroepithelium of the inner ear in mammals, consists of two types of terminally differentiated mechanosensory hair cells and at least four types of supporting cells arrayed precisely along the length of the spiral cochlea. In mice, the progenitors of greater than 80% of both hair cells and supporting cells undergo their terminal division between embryonic day 13 (E13) and E14. As in humans, these cells persist in a non-proliferative state throughout the adult life of the animal. Here we report that the correct timing of cell cycle withdrawal in the developing organ of Corti requires p27(Kip1), a cyclin-dependent kinase inhibitor that functions as an inhibitor of cell cycle progression. p27(Kip1) expression is induced in the primordial organ of Corti between E12 and E14, correlating with the cessation of cell division of the progenitors of the hair cells and supporting cells. In wild-type animals, p27(Kip1) expression is downregulated during subsequent hair cell differentiation, but it persists at high levels in differentiated supporting cells of the mature organ of Corti. In mice with a targeted deletion of the p27(Kip1) gene, proliferation of the sensory cell progenitors continues after E14, leading to the appearance of supernumerary hair cells and supporting cells. In the absence of p27(Kip1), mitotically active cells are still observed in the organ of Corti of postnatal day 6 animals, suggesting that the persistence of p27(Kip1) expression in mature supporting cells may contribute to the maintenance of quiescence in this tissue and, possibly, to its inability to regenerate. Homozygous mutant mice are severely hearing impaired. Thus, p27(Kip1) provides a link between developmental control of cell proliferation and the morphological development of the inner ear.

Animals↗
PubMed2006

Streptozotocin-induced partial beta cell depletion in nude mice without hyperglycaemia induces pancreatic morphogenesis in transplanted embryonic stem cells.

AIMS/HYPOTHESIS: It appears that the adult pancreas has limited regenerative ability following beta cell destruction by streptozotocin (STZ). However, it is not clear if this limitation is due to an inability to respond to, rather than an absence of, regenerative stimuli. In this study we aimed to uncouple the regenerative signal from the regenerative response by using an exogenous stem cell source to detect regenerative stimuli produced by the STZ-injured pancreas at physiological blood glucose levels. METHOD: Adult nude mice received 150 mg/kg STZ and 1x10(6) J1 mouse embryonic stem (ES) cells by i.p. injection. Permanent beta cell depletion of 50% was estimated from the ratio of beta:alpha cells in pancreata from STZ-treated mice compared with control animals after 24 days. RESULTS: Transplanted ES cells homed to the STZ-injured pancreas and formed tumours. Immunocytochemical analysis of pancreas-associated ES tumours revealed foci containing insulin/PDX-1 double-positive and glucagon-positive/PDX-1-negative cell clusters associated with PDX-1-positive columnar lumenal epithelium and extensive alpha-amylase-positive pancreatic acini comprising approximately 0.1% of ES tumour volume. CONCLUSIONS/INTERPRETATION: These data indicate that (1) the adult pancreas produces a milieu of regenerative stimuli following beta cell destruction, and (2) this is not dependent on hyperglycaemic conditions; (3) these regenerative stimuli appear to recapitulate the signalling pathways of embryonic development, since both exocrine and endocrine lineages are produced from PDX-1-positive precursor epithelium. This model will be useful for characterising the regenerative mechanisms in the adult pancreas.

Animals↗
PubMed2001

Spatial and temporal emergence of high proliferative potential hematopoietic precursors during murine embryogenesis.

During mouse embryogenesis, two waves of hematopoietic progenitors originate in the yolk sac. The first wave consists of primitive erythroid progenitors that arise at embryonic day 7.0 (E7.0), whereas the second wave consists of definitive erythroid progenitors that arise at E8.25. To determine whether these unilineage hematopoietic progenitors arise from multipotential precursors, we investigated the kinetics of high proliferative potential colony-forming cells (HPP-CFC), multipotent precursors that give rise to macroscopic colonies when cultured in vitro. No HPP-CFC were found at presomite stages (E6.5-E7.5). Rather, HPP-CFC were detected first at early somite stages (E8.25), exclusively in the yolk sac. HPP-CFC were found subsequently in the bloodstream at higher levels than the remainder of the embryo proper. However, the yolk sac remains the predominant site of HPP-CFC expansion (>100-fold) until the liver begins to serve as the major hematopoietic organ at E11.5. On secondary replating, embryonic HPP-CFC give rise to definitive erythroid and macrophage (but not primitive erythroid) progenitors. Our findings support the hypothesis that definitive but not primitive hematopoietic progenitors originate from yolk sac-derived HPP-CFC during late gastrulation.

Animals↗
PubMed1992

[Function, molecular structure and gene expression of fibroblast growth factor (FGF/HBGF)].

The structure and function of members of the fibroblast growth factor (FGF) gene family and their receptors are reviewed. All the member of this gene family bind heparin, and therefore, are also called the as heparin-binding growth factor (HBGF). In this review, the structural features of FGF/HBGF are summarized first, and general features of the structure and function of their receptors are then described briefly. After biological effects of FGF/HBGF on adult-type tissues and cultured cells are reviewed, effects of FGF on the mesodermal induction in amphibian (Xenopus laevis) embryonic system are reviewed in more detail. Emphasis is given on the experiments with Xenopus animal cap assay system and also on the injection into Xenopus fertilized eggs of mRNA of dominant defect mutant of FGF receptor, which leads to the formation of embryos with abnormal axial mesoderm.

Animals↗
PubMed1999

The angiotensin type 2 receptor: variations on an enigmatic theme.

Since its discovery and molecular characterization, the angiotensin AT2.receptor has been enigmatic with respect to signalling pathways and function. Evidence now emerges that angiotensin II exerts actions through the AT2 receptor which are directly opposed to those mediated by the AT1 receptor. This can be exemplified e.g. by mutually antagonizing effects on cell growth. Upregulated by the endogenous agonist itself, as well as by several growth- and differentiating factors in development and tissue injury, the AT2 receptor appears to act as a modulator of complex biological programmes involved in embryonic development, cell differentiation, tissue protection and regeneration, as well as in programmed cell death. Research on the AT2 receptor has thus unveiled hitherto unknown functions of the renin-angiotensin system extending far beyond the classical role of this old hormonal system in cardiovascular control.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1997

Mouse gastrulation: the formation of a mammalian body plan.

The process of gastrulation is a pivotal step in the formation of the vertebrate body plan. The primary function of gastrulation is the correct placement of precursor tissues for subsequent morphogenesis. There is now mounting evidence that the body plan is established through inductive interactions between germ layer tissues and by the global patterning activity emanating from embryonic organizers. An increasing number of mouse mutants have been described that have gastrulation defects, providing important insights into the molecular mechanisms that regulate this complex process. In this review, we explore the mouse embryo before and during gastrulation, highlighting its similarities with other vertebrate embryos and its unique characteristics.

Animals↗
PubMed1995

[New parameters for prediction of pathological stage in clinical stage I non-seminomatous testicular tumors].

Traditional histopathological risk factors have failed to predict pathological stage accurately in clinical stage I nonseminomatous testicular germ cell tumours. Histopathology, flow cytometry, cytophotometry, and immunohistochemical staining techniques were used in an effort to define high- and low-risk groups for occult metastasis in a consecutive series of 105 patients who underwent retroperitoneal lymph node dissection. After multiple logistic regression analysis, the proliferative S + G2M cell cycle fraction of the aneuploid tumour stemline was the most highly predictive parameter of pathological stage (P = 0.0004). Using a cut-off of 41%, pathological stage II patients were predicted with a sensitivity of 71%. There were 61 patients with S + G2M values below 41%, and 43 of them had pathological stage I disease (negative predictive value 87%). A low volume of embryonal carcinoma was predominant in low-risk patients, and MIB-1 immunohistochemical staining identified a subgroup of 23% of patients with pathological stage I disease and at extremely low risk of metastatic disease. Assessment of tumour cell proliferation does not allow accurate classification of high-risk patients at a level that is adequate for clinical application. Patients who are at low risk of metastasis, however, can be identified by flow cytometry, immunohistochemical proliferation markers and volume of embryonal carcinoma with 90% certainty. These parameters deserve further study, since identification of a subgroup of patients at extremely low risk of metastasis could potentially reduce the overall morbidity in the management of clinical stage I nonseminomatous testis cancer.

Animals↗
PubMed1989

Contrasting patterns of myc and N-myc expression during gastrulation of the mouse embryo.

myc and N-myc are related genes whose similar protein products may be used for different purposes in vertebrate organisms. We have explored this possibility by using hybridization in situ to examine the expression of myc and N-myc during gastrulation of mouse embryos. Throughout gastrulation, myc RNA was most abundant in extraembryonic cells; by contrast, N-myc RNA was found at highest levels in the expanding primitive streak and other portions of the embryonic mesoderm. Differentiation of mesoderm to epithelioid cells was accompanied by diminished expression of N-myc. Expression of myc was not an inevitable correlate of cellular proliferation. Instead, the gene appeared to be regulated in concert with changes that affect a diversity of cellular properties, including proliferation, invasiveness, and differentiation.

Animals↗
PubMed1999

[Receptors and development of endothelial and hematopoietic cells].

During vertebrate embryonic development, the endothelial and hemopoietic systems are the first system to be specified. In this review, we will summarize recent findings about the molecular mechanisms responsible for the successive steps of the development of these systems: the differentiation of mesodermal cells to endothelial and hemopoietic cells, their proliferation and their interactions to form the vascular system.

Animals↗
PubMed1998

Mutations in genes encoding extracellular matrix proteins suppress the emb-5 gastrulation defect in Caenorhabditis elegans.

The second division of the gut precursor E cells is lethally accelerated during Caenorhabditis elegans gastrulation by mutations in the emb-5 gene, which encodes a presumed nuclear protein. We have isolated suppressor mutations of the temperature-sensitive allele emb-5(hc61), screened for them among dpy and other mutations routinely used as genetic markers, and identified eight emb-5 suppressor genes. Of these eight suppressor genes, at least four encode extracellular matrix proteins, i.e., three collagens and one proteoglycan. The suppression of the emb-5 gastrulation defect seemed to require the maternal expression of the suppressors. Phenotypically, the suppressors by themselves slowed down early embryonic cell divisions and corrected the abnormal cell-division sequence of emb-5 mutant embryos. We propose an indirect stress-response mechanism to be the main cause of the suppression because: (1) none of these suppressors is specific, either to particular temperature-sensitive emb-5 alleles or to the emb-5 gene; (2) suppressible alleles of genes, reported here or elsewhere, are temperature sensitive or weak; (3) the suppression is not strong but marginal; (4) the suppression itself shows some degree of temperature dependency; and (5) none of the extracellular matrix proteins identified here is known to be expressed in oocytes or early embryos, despite the present observation that the suppression is maternal.

Alleles↗