pinecone/core-2020-05-10-deduplication huggingface dataset:
Muscle-Specific Cell Ablation Conditional upon Cre-Mediated DNA Recombination in Transgenic Mice Leads to Massive Spinal and Cranial Motoneuron Loss

AbstractWe describe here a binary transgenic system based on Cre-mediated DNA recombination for genetic cell ablation in mice that enabled us to obtain skeletal muscle-deficient embryos by mating two phenotypically normal transgenic lines. In those embryos, skeletal muscles are eliminated as a consequence of the expression of the gene encoding the diphtheria toxin A fragment. Cell ablation occurs gradually beginning approximately on embryonic day (E) 12.5, and by E18.5 almost all skeletal muscle is absent. Analysis of the consequences of muscle cell ablation revealed that almost all spinal motoneurons are lost by E18.5, providing strong evidence that survival of spinal motoneurons during embryogenesis is dependent on signals from their target tissue, skeletal muscle, and that trophic signals produced by nonmuscle sources are sufficient to support survival of no more than 10% of embryonic spinal motoneurons in the absence of muscle-derived signals. There was also substantial loss of cranial (hypoglossal and facial) motoneurons in the muscle-deficient embryos, thus indicating that cranial motoneuron survival is also dependent on trophic signals produced by their target tissue. Although spinal motoneurons are a major target of spinal interneurons, the loss of motoneurons did not affect interneuron survival. Muscle-deficient embryos had a cleft palate and abnormalities of the lower jaw, raising the possibility that they might serve as a mouse model for the human disorder, Robin sequence. The data reported here demonstrate the utility of a binary transgenic system for obtaining mouse embryos in which a specific cell population has been ablated, so that its role in embryonic development can be studied
Bone Morphogenetic Proteins Induce Apoptosis and Growth Factor Dependence of Cultured Sympathoadrenal Progenitor Cells

AbstractNeuron numbers in developing vertebrate organisms are regulated by the availability of growth factors which promote their survival. However, neuron survival may also be regulated by growth factors which promote rather than prevent cell death. This study examined the effects of bone morphogenetic proteins (BMPs) in inducing apoptosis of MAH cells, an immortalized sympathoadrenal progenitor cell line. Treatment of MAH cells with BMP2 or BMP4 killed the cells in a dose-dependent manner. By contrast, treatment with BMP7 or TGFβ1 failed to affect survival, suggesting that induction of apoptosis is specific to the dpp subgroup of BMPs. Survival after treatment with BMP2 or BMP4 required addition of fibroblast growth factor (FGF) and nerve growth factor (NGF), indicating that BMP treatment made the neurons dependent upon an exogenous factor for survival. Several experimental observations suggested an apoptotic mechanism for BMP-induced death. After BMP2 treatment, the cells progressively shrank and became pyknotic. Further, there was prominent endonucleosomic cleavage of DNA (laddering) as well as TUNEL staining. Moreover, BMP-induced death was inhibited by the caspase inhibitor z-VAD and was partially prevented by the endonuclease inhibitor aurintricarboxylic acid. These observations suggest that neuron numbers may be regulated by factors which promote death and that exposure to such factors may be a signal for the development of dependence upon other growth factors for survival
Cell Death during Development of Testis and Cerebellum in the Mutant MouseWeaver

AbstractThe murine mutationweaverconfers early death during development on cells in testes, cerebellum, and midbrain. The results reported here support the hypothesis that the action ofweaveris intrinsic to testes and independent of Sertoli cells: germ cells are the only testicular cell type seen to die in weaver homozygotes, while Sertoli cell-dependent development of the blood testis barrier is normal. This report includes characterization of patterns of germ cell death and cerebellar granule cell death in homozygous weavers with respect to that seen during normal development byin situend-labeling of DNA and high-magnification light microscopy. Comparison of the spatial distribution of dying cells in the weaver's cerebellum with that of dividing cells revealed disarray in the external germinal zone. The results show that cells vulnerable toweaverdie by apoptotic and nonapoptotic mechanisms and indicate thatweaver-induced cell death is not the consequence of extended naturally occurring developmental cell death, although their timing overlaps. Thus, although the death of cells in each region is likely to be caused by the same mutation, a base pair substitution in the G protein-coupled inwardly rectifying potassium channel 2 gene, the cell death program activated differs depending on cell type
Local Inhibitory Action of BMPs and Their Relationships with Activators in Feather Formation: Implications for Periodic Patterning

AbstractThe formation of periodic patterns is fundamental in biology. Theoretical models describing these phenomena have been proposed for feather patterning; however, no molecular candidates have been identified. Here we show that the feather tract is initiated by a continuous stripe ofShh, Fgf-4,andPtcexpression in the epithelium, which then segregates into discrete feather primordia that are more stronglyShhandFgf-4positive. The primordia also becomeBmp-2andBmp-4positive. Bead-mediated delivery of BMPs inhibits local feather formation in contrast with the activators, SHH and FGF-4, which induce feather formation. Both FGF-4 and SHH induce local expression ofBmp-4,while BMP-4 suppresses local expression of both. FGF-4 also inducesShh.Based on these findings, we propose a model that involves (1) homogeneously distributed global activators that define the field, (2) a position-dependent activator of competence that propagates across the field, and (3) local activators and inhibitors triggered in sites of individual primordia that act in a reaction–diffusion mechanism. A computer simulation model for feather pattern formation is also presented
Expression of a Constitutively Active Type I BMP Receptor Using a Retroviral Vector Promotes the Development of Adrenergic Cells in Neural Crest Cultures

AbstractPrevious work has demonstrated that the bone morphogenetic proteins (BMP)-2, BMP-4, and BMP-7 can promote the development of tyrosine hydroxylase (TH)-positive and catecholamine-positive cells in quail trunk neural crest cultures. In the present work, we showed that mRNA for the type I bone morphogenetic protein receptor IA (BMPR-IA) was present in neural crest cells grown in the absence or presence of BMP-4. We have used a replication-competent avian retrovirus to express a constitutively active form of BMPR-IA in neural crest cells in culture. Cultures grown in the absence of BMP-4 and infected with retrovirus containing a construct encoding this activated BMPR-IA developed five times more TH-immunoreactive and catecholamine-positive cells than uninfected control cultures or cultures infected with virus bearing the wild-type BMPR-IA cDNA. The number of TH-positive cells which developed was dependent on the concentration of virus bearing the activated receptor cDNA used in the experiments. Most TH-positive cells which developed also contained viral p19 protein. Total cell number was not affected by infection with the virus containing the activated receptor construct. The effect of the activated receptor was phenotype-specific since infection with the virus bearing the activated receptor cDNA did not alter the number or morphology of microtubule-associated protein (MAP)2-immunoreactive cells, which are distinct from the TH-positive cell population. These findings are consistent with the observation that MAP2-positive cells are not affected by the presence of BMP-4. Taken together, these results suggest that activity of BMPR-IA is an important element in promoting the development of the adrenergic phenotype in neural crest cultures
The Hybrid Histidine KinasedhkBRegulates Spore Germination inDictyostelium discoideum

AbstractSpore germination is a defined developmental process that marks a critical point in the life cycle ofDictyostelium discoideum.Upon germination the environmental conditions must be conducive to cell growth to ensure survival of emerged amoebae. However, the signal transduction pathways controlling the various aspects of spore germination in large part remain to be elucidated. We have used degenerate PCR to identifydhkB,a two-component histidine kinase, fromD. discoideum. DhkBis predicted to be a transmembrane hybrid sensor kinase. ThedhkB-null cells develop with normal timing to give what seem to be mature fruiting bodies by 22 to 24 h. However, over the next several hours, the ellipsoidal and encapsulated spores proceed to swell and germinatein situwithin the sorus and thus do not respond to the normal inhibitors of germination present within the sorus. The emerged amoebae dehydrate due to the high osmolarity within the sorus, and by 72 h 4% or less of the amoebae remain as spores, while most cells are now nonviable. Precocious germination is suppressed by ectopic activation of or expression of cAMP-dependent protein kinase A. Additionally, at 24 h the intracellular concentration of cAMP ofdhkB−spores is 40% that ofdhkB+spores. The results indicate that DHKB regulates spore germination, and a functional DHKB sensor kinase is required for the maintenance of spore dormancy. DHKB probably acts by maintaining an active PKA that in turn is inhibitory to germination
The Cell Cycle Program in Germ Cells of theDrosophilaEmbryo

AbstractThe germ cells of metazoans follow a program of proliferation that is distinct from proliferation programs of somatic cells. Despite their developmental importance, the cell proliferation program in the metazoan primordial germ cells is not well characterized and the regulatory controls are not understood. InDrosophila melanogaster,germ cell precursors (called pole cells) proliferate early in embryogenesis and then enter a prolonged quiescence. We found that polar nuclear divisions are asynchronous and lag behind somatic nuclear divisions during syncytial cycles 9 and 10. Thus, the polar division program deviates from the somatic division program when pole nuclei and somatic nuclei still share a common cytoplasm, earlier than previously thought. The lag in polar nuclear divisions is independent ofgrapes,which is required for lengthening somatic cell cycles 10–13. Mapping of the last S phase in pole cells and measurement of their DNA content indicate that pole cells become quiescent in G2 phase of the cell cycle. We were able to drive quiescent pole cells into mitosis by induction of either an activator of Cdc2 (Cdc25stringphosphatase) or a mutant form of Cdc2 that cannot be inhibited by phosphorylation. In contrast, induction of wild-type Cdc2 with a mitotic cyclin did not induce mitosis in pole cells. We propose that inhibition of Cdc2 by phosphorylation contributes to G2 arrest in pole cells during embryogenesis. Furthermore, pole cells enter G1 following induced mitoses, indicating that entry into both mitosis and S phase is blocked in quiescent pole cells. These studies represent the first molecular characterization of proliferation in embryonic germ cells of Drosophil
Multiple Digit Formation inXenopusLimb Bud Recombinants

AbstractWe prepared recombinant limb buds ofXenopustadpoles by grafting a mesenchyme mass of the hindlimb bud. TheXenopusrecombinant limb buds with dissociated and reaggregated mesenchyme developed more than 30 digits with cartilage segmentation, while those with undissociated mesenchyme developed a limb with normal cartilage pattern. Before the formation of multiple digits, a patchy expression pattern offgf-8,an AER marker, was observed in the distal region of recombinant limb buds.shh,a ZPA (zone of polarizing activity) marker, was expressed broadly in the distal region of recombinants. Recombinant limb buds with the reaggregated mesenchyme of anterior halves formed anterior digits with claws, and those with the mesenchyme of posterior halves formed posterior digits without claws. The temporal and spatial changes in the potency of multiple digit formation are discussed with reference to the regenerative capacity ofXenopuslimb buds
Matrix Metalloproteinase Inhibitors Disrupt Spicule Formation by Primary Mesenchyme Cells in the Sea Urchin Embryo

AbstractThe primary mesenchyme cells of the sea urchin embryo construct an elaborate calcareous endoskeletal spicule beginning at gastrulation. This process begins by ingression of prospective primary mesenchyme cells into the blastocoel, after which they migrate and then fuse to form a syncytium. Skeleton deposition occurs in spaces enclosed by the cytoplasmic cables between the cell bodies. Experiments are described which probe the role of proteases in these early events of spicule formation and their role in the continued elaboration of the spicule during later stages of embryogenesis. We find that several inhibitors of metalloproteinases inhibit the continuation of spiculogenesis, an effect first reported by Roeet al.(Exp. Cell Res.181, 542–550, 1989). A detailed study of one of these inhibitors, BB-94, shows that fusion of primary mesenchyme cells still occurs in the presence of the inhibitor and the formation of the first calcite granule is not impeded. Continued elaboration of the spicule, however, is completely stopped; addition of the inhibitor during the active elongation of the spicule stops further elongation immediately. Removal of the inhibitor allows resumption of spicule growth. The inhibition is accompanied by almost complete cessation of massive Ca ion transport via the primary mesenchyme cells to the spicule. The inhibitor does not prevent the continued synthesis of several spicule matrix proteins. Electron microscopic examination of inhibited primary mesenchyme cells shows an accumulation of characteristic vesicles in the cytoplasm. Gel zymography demonstrates that although most proteases in homogenates of primary mesenchyme cells are not sensitive to the inhibitorin vitro,a protease of low abundance detectable in the medium of cultured primary mesenchyme cells is inhibited by BB-94. We propose that the inhibitor is interfering with the delivery of precipitated calcium carbonate and matrix proteins to the site(s) of spicule growth
Specific Arrest of Cardiogenesis in Cultured Embryonic Stem Cells Lacking Cripto-1

AbstractThe molecular events of cardiac lineage specification and differentiation are largely unknown. Here we describe the involvement of a growth factor with an EGF-like domain, Cripto-1 (Cr-1), in cardiac differentiation. During embryonic development, Cr-1 is expressed in the mouse blastocyst, primitive streak, and later is restricted to the developing heart. To investigate the role of Cr-1, we have generated Cr-1-negative embryonic stem (ES) cell lines by homologous recombination. The resulting double “knockout” ES cells have selectively lost the ability to form beating cardiac myocytes, a process that can be rescued by reintroducing Cr-1 gene back into the Cr(−/−) cells. Furthermore, the lack of functional Cr-1 is correlated with absence of expression of cardiac-specific myosin light and heavy chain genes during differentiation. Differentiation into other cell types including skeletal muscle is not disrupted. These results suggest that Cr-1 is essential for contractile cardiomyocyte formationin vitro
Notch4 and Wnt-1 Proteins Function to Regulate Branching Morphogenesis of Mammary Epithelial Cells in an Opposing Fashion

AbstractElongation and branching of epithelial ducts is a crucial event during the development of the mammary gland. Branching morphogenesis of the mouse mammary epithelial TAC-2 cell line was used as an assay to examine the role of Wnt, HGF, TGF-β, and the Notch receptors in branching morphogenesis. Wnt-1 was found to induce the elongation and branching of epithelial tubules, like HGF and TGF-β2, and to strongly cooperate with either HGF or TGF-β2 in this activity. Wnt-1 displayed morphogenetic activity in TAC-2 cells as it induced branching even under conditions that normally promote cyst formation. The Notch4(int-3) mammary oncoprotein, an activated form of the Notch4 receptor, inhibited the branching morphogenesis normally induced by HGF and TGF-β2. The minimal domain within the Notch4(int-3) protein required to inhibit morphogenesis consists of the CBF-1 interaction domain and the cdc10 repeat domain. Coexpression of Wnt-1 and Notch4(int-3) demonstrates that Wnt-1 can overcome the Notch-mediated inhibition of branching morphogenesis. These data suggest that Wnt and Notch signaling may play opposite roles in mammary gland development, a finding consistent with the convergence of the wingless and Notch signaling pathways found inDrosophila
Growth Promoting Activity of Oocytes on Granulosa Cells Is Decreased upon Meiotic Maturation

AbstractAn increasing body of evidence indicates that the oocyte plays an active role in the control of ovarian follicle development in mammals. In the present study, we have examined the role of oocytes in regulating granulosa cell proliferation. Rat and bovine oocytes cocultured with rat granulosa cells stimulated granulosa cell DNA synthesis and DNA content in the cultures. FSH or cAMP further amplified this effect. Poor-quality oocytes showed a marked decrease in their stimulatory effect. Stimulation of DNA synthesis by bovine oocytes seems to be cell-type specific, since Swiss 3T3 fibroblasts and CCL-64 mink lung epithelial cells were not responsive, while primary cultures of rat and bovine granulosa cells and the bovine granulosa cell line BGC-1 showed significant responses. Oocyte-conditioned medium produced only a slight stimulation of rat granulosa cell DNA synthesis. However, the effect of oocyte coculture was dependent on the total incubation volume, suggesting that the growth promoting activity was mediated by a soluble factor. The stimulation elicited by bovine oocytes was evident even in the presence of maximally effective doses of transforming growth factor-β or tumor necrosis factor-α, indicating that neither of these growth factors was responsible for this effect.In vitromaturation of bovine oocytes was associated with a marked decrease in the stimulatory activity. This decrease was partially prevented when maturation was blocked by addition of cycloheximide. Comparison of the developmental pattern of the secretion of the growth promoting activity with that of the cumulus expansion-enabling factor indicated that both activities can be dissociated. Our data suggest the existence of a very labile factor produced by the oocyte before completion of the first meiotic division that promotes granulosa cell proliferation
Ectoderm Cell–ECM Interaction Is Essential for Sea Urchin Embryo Skeletogenesis

AbstractParacentrotus lividussea urchin nectin (Pl-nectin) is an extracellular matrix (ECM) protein of the sea urchin embryo on the apical surface of the ectoderm and has been shown to be an adhesive substrate for embryonic cells. A monoclonal antibody (McAb) toPl-nectin was generated that inhibits the adhesion of blastula cells toPl-nectin-coated substrates in anin vitrofunctional assay. To examine for possiblein vivofunctions ofPl-nectin, Fab fragments (Fabs) ofPl-nectin McAb were added to early blastulae. Ingression of primary mesenchyme cells was not affected by Fabs. As control embryos reached the pluteus stage, treated embryos showed a severe inhibition of skeletal elongation and patterning. When the Fabs were injected directly into the blastocoel, even at higher concentration than was applied externally, skeletogenesis was normal. Therefore, the effect of the antibody on spiculogenesis was indirect. The treatment was partially reversible as embryos eventually seemed to recover and elongate spicules, although with an incorrect patterning. Migration of pigment cells was also affected by the Fabs, since they did not disperse throughout the ectoderm but remained clustered in ectopic areas. In contrast, the development of endoderm structures was not affected. Our results indicate that in the sea urchin embryo the appropriate contact of ectodermal cells with outer ECM components is essential for the correct morphogenesis of inner mesodermal structures
Requirement for Brn-3b in Early Differentiation of Postmitotic Retinal Ganglion Cell Precursors

AbstractThe Brn-3 subfamily of POU domain transcription factors consists of Brn-3a, -3b, and -3c, which are important regulators for sensorineural development. Despite the expression of all three factors in retinal ganglion cells, earlier studies have shown thatBrn-3bis the only one among the threeBrn-3genes that is essential for development of ∼70% of ganglion cells in the murine retina. I report here that Brn-3b displays a spatiotemporal expression pattern characteristic of the dynamic profile of ganglion cell genesis during murine retinal development. Moreover, it is initially turned on in postmitotic ganglion cell precursors 2 days before the onset of Brn-3a and -3c expression in differentiated ganglion cells. During the entire period of retinal ganglion cell genesis, the postmitotic ganglion cell precursors that would normally become Brn-3b+cells fail to properly differentiate inBrn-3b−/−mice, as evidenced by a twofold reduction in the optic nerve size and diminished expression of several ganglion cell markers. The undifferentiated ganglion cell precursors appear to be degenerated by apoptosis within the ganglion cell layer during the perinatal and early postnatal period. I propose that retinal ganglion cells develop following two separate differentiation pathways—Brn-3b dependent and Brn-3b independent. In the Brn-3b-dependent mechanism, Brn-3b may be required to initiate a particular differentiation program for a large set of postmitotic ganglion precursors to properly differentiate into the 70%, Brn-3b-dependent retinal ganglion cells
Components of the Signaling Pathway Linking the 1-Methyladenine Receptor to MPF Activation and Maturation in Starfish Oocytes

AbstractStarfish oocytes are arrested at the G2/M-phase border of meiosis I. Exposure to their natural mitogen, 1-methyladenine (1-MA), leads to the activation of MPF and MAP kinase, resumption of the meiotic cell cycle, and fertilization competency. The 1-MA receptor has not yet been identified, but it is known to be linked functionally to a pertussis toxin-sensitive G-protein. Gβγ appears to be the major effector of the 1-MA receptor, since injection of Gβγ, but not activated Gαi, leads to the activation of MPF, entry into meiosis, and oocyte maturation. The components that connect Gβγ to MPF and MAP kinase activation in oocytes are unknown. In mammalian cells, a novel phosphatidylinositol 3-kinase, PI-3 kinase-γ, links Gβγ to the MAP kinase activation pathway. Here we show that PI-3 kinase is required for starfish oocyte maturation. LY294002 and wortmannin, inhibitors of PI-3 kinase, block MPF and MAP kinase activation and entry into meiosis. Inhibition by LY294002 is reversible and limited to the hormone-dependent period. Neither inhibitor, however, blocks the earliest hormone-induced event, formation of actin spikes at the cell membrane. By contrast, pertussis toxin blocks both actin spiking and later events, arguing that PI-3 kinase functions downstream of Gβγ. Finally, we show that unlike the well-studied case inXenopusoocytes, where MAP kinase is an essential component of the MPF activation pathway, MAP kinase is not required for either MPF activation or subsequent oocyte maturation in starfish. Instead, its major role appears to be suppression of DNA synthesis in unfertilized, haploid eggs