bluuebunny/crossref_metadata_2025_split huggingface dataset:
Control of segmental asymmetry in Drosophila embryos
Authors: Manoukian Armen S. , Krause Henry M. 
Published: April 2021

ABSTRACT
During Drosophila development, an important aspect of body patterning is the division of the embryo into repeating morphological units referred to as parasegments. The parasegmental domains are first defined at the blastoderm stage by alternating stripes of transcripts encoded by the pair-rule genes fushi tarazu (ftz) and even-skipped (eve) and later by stripes encoded by the segment polarity genes engrailed (en) and wingless. Here, we show that the runt gene (run) is required to generate asymmetries within these parasegmental domains. Using a heat-shock-inducible run transgene, we found that ectopic run expression leads to rapid repression of eve stripes and a somewhat delayed expansion of ftz stripes. Unexpectedly, we also found that ectopic run was a rapid and potent repressor of odd-numbered en stripes. Two remarkably different segmental phenotypes were generated as a consequence of these effects. In solving the mechanisms underlying these phenotypes, we discovered that the positioning of en stripes is largely determined by the actions of negative regulators. Our data indicate that run is required to limit the domains of en expression in the odd-numbered parasegments, while the odd-skipped gene is required to limit the domains of en expression in the even-numbered parasegments. Activation of en at the anterior margins of both sets of parasegments requires the repression of run and odd by the product of the eve gene. The spatial restriction of gene expression via negative and double negative pathways such as these is likely to be a common theme during development.
Induction of cardiac muscle differentiation in isolated animal pole explants of Xenopus laevis embryos
Authors: Logan Malcolm , Mohun Tim 
Published: April 2021

ABSTRACT
We have isolated a cDNA fragment encoding a portion of the myosin heavy chain α-isoform (XMHCα) in the amphibian, Xenopus laevis. The XMHCα transcript is highly enriched in adult heart RNA and is expressed exclusively in embryonic heart tissue. It therefore provides a tissue-specific marker for cardiac muscle differentiation during early embryogenesis. Using an RNAase protection assay, we can detect the onset of cardiac muscle differentiation in an anterior, ventral region of tailbud embryos, many hours before the appearance of a beating heart. Whole-mount in situ RNA hybridisation indicates that expression of the XMHCα gene is restricted to the developing heart pri-mordium. XMHCα gene expression can also be induced in isolated animal pole explants of blastulae by treatment with the growth factor, activin A. Induction is dose-dependent, requiring high doses of the growth factor compared with that required for myotomal (skeletal) muscle differentiation. In contrast, no XMHCα tran-scripts are detected in explants incubated with basic FGF, despite the induction of myotomal muscle differ-entiation. Activin-induced explants show a similar temporal pattern of XMHCα gene expression to that found in normal embryogenesis. Furthermore, cells expressing this gene appear clustered in one or two foci within fused explant aggregates, which often show regular, spontaneous contractions after several days in culture. These results show that terminal differentiation of cardiac muscle can occur in growth factor-induced explants and may be distinguished from skeletal muscle differentiation by the dose and nature of the inducing factor.
Neurogenesis in the insect brain: cellular identification and molecular characterization of brain neuroblasts in the grasshopper embryo
Authors: Zacharias David , Leslie J. , Williams D. , Meier Thomas , Reichert Heinrich 
Published: April 2021

ABSTRACT
Brain neuroblasts in the embryonic grasshopper were studied by toluidine blue staining, BrdU incorporation, and immunocytochemistry in whole-mounts as well as by reconstruction of stained serial sections. Large dividing neuroblasts are observed by the 25% stage. During early neurogenesis these neuroblasts generate their progeny through mechanisms similar to those that occur in the segmental ganglia; each neuroblast divides asymmetrically to produce a chain of ganglion mother cells, and each ganglion mother cell divides symmetrically to produce a pair of neurons. Approximately 130 mitotically active, large neuroblasts are found in each brain hemisphere at the 30–45% stages. Through morphogenetic movements that occur between the 30–35% stages these neuroblasts become located in positions which are predictive of the major brain regions that they give rise to. Many of the brain neuroblasts can be identified as individuals based on their stereotyped position in the neurogenic array. Immunocytochemical experiments with antibodies against, engrailed, fasciclin I and TERM-1 show that brain neuroblasts can also be characterized by their expression of cell-specific molecular labels. These studies indicate that many features of the complex mature insect brain derive from a surprisingly simple and stereotyped set of neuronal precursor cells. Thus, many of the concepts and methods that have been used to study neurogenesis in the simpler segmental ganglia may also be applicable to the insect brain.
Different neurotrophins are expressed and act in a developmental sequence to promote the survival of embryonic sensory neurons
Authors: Buchman Vladimir L. , Davies Alun M. 
Published: April 2021

ABSTRACT
To investigate if different neurotrophins regulate the survival of neurons at successive developmental stages, we studied the effect of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neu-rotrophin-3 (NT-3) on the survival of mouse trigeminal neurons at closely staged intervals in development. We show that during the earliest stages of target field innervation trigeminal neurons display a transitory survival response to BDNF and NT-3. This response is lost as the neurons become NGF-dependent shortly before neuronal death begins in the trigeminal ganglion. BDNF and NT-3 mRNAs are expressed in the peripheral trigeminal target field before the arrival of the earliest axons and the onset of NGF mRNA expression. The levels of BDNF and NT-3 mRNAs peak during the early stages of target field innervation and decline shortly after the loss of neuronal responsiveness to BDNF and NT-3. Our study provides the first clear evidence that different target-derived neurotrophins can act sequentially to promote the survival of developing neurons.
Regulative capacity of the cranial neural tube to form neural crest
Authors: Scherson Talma , Serbedzija George , Fraser Scott , Bronner-Fraser Marianne 
Published: April 2021

ABSTRACT
In avian embryos, cranial neural crest cells emigrate from the dorsal midline of the neural tube shortly after neural tube closure. Previous lineage analyses suggest that the neural crest is not a pre-segregated population of cells within the neural tube; instead, a single progenitor in the dorsal neural tube can contribute to neurons in both the central and the peripheral nervous systems (Bronner-Fraser and Fraser, 1989Neuron 3, 755–766). To explore the relationship between the ‘premigratory’ neural crest cells and the balance of the cells in the neural tube in the midbrain and hindbrain region, we have challenged the fate of these populations by ablating the neural crest either alone or in combination with the adjoining ventral portions of the neural tube. Focal injections of the vital dye, DiI, into the neural tissue bordering the ablated region demonstrate that cells at the same axial level, in the lateral and ventral neural tube, regulate to reconstitute a population of neural crest cells. These cells emigrate from the neural tube, migrate along normal pathways according to their axial level of origin and appear to give rise to a normal range of derivatives. This regulation following ablation suggests that neural tube cells normally destined to form CNS derivatives can adjust their prospective fates to form PNS and other neural crest derivatives until 4.5–6 hours after the time of normal onset of emigration from the neural tube.
Loss of Hox-A1 (Hox-1.6) function results in the reorganization of the murine hindbrain
Authors: Carpenter Ellen M. , Goddard Judy M. , Chisaka Osamu , Manley Nancy R. , Capecchi Mario R. 
Published: April 2021

ABSTRACT
Targeted disruption of the murine hox-A1 gene results in severe defects in the formation of the hindbrain and associated cranial ganglia and nerves. Carbocyanine dye injections were used to trace afferent and efferent projections to and from the hindbrain in hox-A1 /hox-A1 mutant mice. Defects were observed in the position of efferent neurons in the hindbrain and in their projection patterns. In situ hybridization was used to analyze the transcription pattern of genes expressed within specific rhombomeres. Krox-20, int-2 (fgf-3), and hox-B1 all display aberrant patterns of expression in hox-A1 mutant embryos. The observed morphological and molecular defects suggest that there are changes in the formation of the hindbrain extending from rhombomere 3 through rhombomere 8 including the absence of rhom-bomere 5. Also, motor neurons identified by their axon projection patterns which would normally be present in the missing rhombomere appear to be respecified to or migrate into adjacent rhombomeres, suggesting a role for hox-A1 in the specification of cell identity and/or cell migration in the hindbrain.
Expression of Synaptotagmin in Drosophila reveals transport and localization of synaptic vesicles to the synapse
Authors: Littleton J. Troy , Bellen Hugo J. , Perin Mark S. 
Published: April 2021

ABSTRACT
Synaptotagmin is a synaptic vesicle-specific integral membrane protein that has been suggested to play a key role in synaptic vesicle docking and fusion. By monitoring Synaptotagmin’s cellular and subcellular distribution during development, it is possible to study synaptic vesicle localization and transport, and synapse formation. We have initiated the study of Synaptotag-min’s expression during Drosophila neurogenesis in order to follow synaptic vesicle movement prior to and during synapse formation, as well as to localize synaptic sites in Drosophila. In situ hybridizations to whole-mount embryos show that synaptotagmin (syt) message is present in the cell bodies of all peripheral nervous system neurons and many, if not all, central nervous system neurons during neurite outgrowth and synapse formation, and in mature neurons. Immunocytochemical staining with antisera specific to Synaptotagmin indicates that the protein is present at all stages of the Drosophila life cycle following germ band retraction. In embryos, Synaptotagmin is only transiently localized to the cell body of neurons and is transported rapidly along axons during axonogenesis. After synapse formation, Synaptotagmin accumulates in a punctate pattern at all identifiable synaptic contact sites, suggesting a general role for Synaptotagmin in synapse function. In embryos and larvae, the most intense staining is found along two broad longitudinal tracts on the dorsal side of the ventral nerve cord and the brain, and at neuromuscular junctions in the periphery. In the adult head, Synaptotagmin localizes to discrete regions of the neurophil where synapses are predicted to occur. These data indicate that synaptic vesicles are present in axons before synapse formation, and become restricted to synaptic contact sites after synapses are formed. Since a similar expression pattern of Synaptotagmin has been reported in mammals, we propose that the function of Synaptotagmin and the mechanisms governing localization of the synaptic vesicle before and after synapse formation are conserved in invertebrate and vertebrate species. The ability to mark synapses in Drosophila should facilitate the study of synapse formation and function, providing a new tool to dissect the molecular mechanisms under-lying these processes.
Stem cell factor and leukemia inhibitory factor promote primordial germ cell survival by suppressing programmed cell death (apoptosis)
Authors: Pesce Maurizio , Farrace Maria Grazia , Piacentini Mauro , Dolci Susanna , Felici Massimo De 
Published: April 2021

ABSTRACT
Proliferating primordial germ cells (PGCs) isolated from mouse embryos soon after their arrival in the genital ridges would only survive in vitro at temperature of less than 30°C (De Felici, M. and McLaren, A. (1983). Exp. Cell. Res. 144, 417 –427; Wabik-Sliz, B. and McLaren, A. (1984). Exp. Cell. Res. 154, 530 –536) or when co-cultured on cell feeder layers (Donovan, P. J., Stott, D., Godin, I., Heasman, J. and Wylie, C. C. (1986). Cell 44, 831–838; De Felici, M. and Dolci, S. (1991). Dev. Biol. 147, 281– 284).
In the present paper we report that mouse PGC death in vitro occurs with all the hallmarks of programmed cell death or apoptosis. We found that after 4–5 hours in culture many PGCs isolated from 12.5 dpc fetal gonads assumed a nuclear morphology and produced membrane bound fragments (apoptotic bodies) typical of apoptotic cells. In addition, PGCs in culture accumulated high level of tissue transglutaminase (tTGase; an enzyme that is induced and activated during apoptosis) and showed extensive degradation of DNA to oligonucleosomal fragments, which is characteristic of apoptosis. The physiological relevance of this mechanism of PGC death is supported by the finding that some PGCs undergoing apoptosis, as revealed by the high level of tTGase expression, were detected in the embryo. Most importantly, we show that the addition of stem cell factor (SCF) or leukemia inhibitory factor (LIF) to the culture medium, two cytokines known to favour PGC survival and/or proliferation in vitro, markedly reduced the occurrence of apoptosis in PGCs during the first hours in culture. These last results suggest a novel mechanism by which these two cytokines may affect the in vitro as well possibly in vivo development of mammalian PGCs.
Selective accumulation of MyoD and Myogenin mRNAs in fast and slow adult skeletal muscle is controlled by innervation and hormones
Authors: Hughes Simon M. , Taylor Jane M. , Tapscott Stephen J. , Gurley Cathy M. , Carter William J. , Peterson Charlotte A. 
Published: April 2021

ABSTRACT
Each of the myogenic helix-loop-helix transcription factors (MyoD, Myogenin, Myf-5, and MRF4) is capable of activating muscle-specific gene expression, yet distinct functions have not been ascribed to the individual proteins. We report here that MyoD and Myogenin mRNAs selectively accumulate in hindlimb muscles of the adult rat that differ in contractile properties: MyoD is prevalent in fast twitch and Myogenin in slow twitch muscles. The distribution of MyoD and Myogenin transcripts also differ within a single muscle and correlate with the proportions of fast glycolytic and slow oxidative muscle fibres, respectively. Furthermore, the expression of a transgene consisting of a muscle-specific cis-regulatory region from the myoD gene controlling lacZ was primarily associated with the fast glycolytic fibres. Alteration of the fast/slow fibre type distribution by thyroid hormone treatment or by cross-reinnervation resulted in a corresponding alteration in the MyoD/Myogenin mRNA expression pattern. These findings show that the expression of specific myogenic helix-loop-helix regulators is under the control of innervation and humoral factors and may mediate differential control of contractile protein gene expression in adult muscle.
Embryonic wing and leg motoneurons have intrinsically different survival properties
Authors: Mettling Clément , Camu William , Henderson Christopher E. 
Published: April 2021

ABSTRACT
Although spinal motoneurons in the chick embryo are born in a rostro-caudal gradient, the timing of their naturally occurring cell death varies in the opposite sense: brachial motoneurons (MNs) die later than lumbar MNs. We used in vitro methods to determine whether this difference results from factors intrinsic or extrinsic to the MNs. Embryonic MNs were purified from E5 lumbar and brachial spinal cord by a method that enriches for MNs with heightened trophic requirements; they were cultured in serum-free medium. In the absence of muscle extract, death of both populations was rapid; similar results are obtained in ovo after limb ablation. In the presence of muscle extracts, however, lumbar MNs always died more rapidly than brachial MNs, as in the normal embryo. We tested the ability of wing and leg bud extracts to provide trophic factors for MNs in culture. MNs from a given level responded equally well to wing or leg extracts. However, dose-response studies showed that near-maximal survival of brachial MNs was obtained with lower concentrations of limb extract than those required for optimal survival of lumbar MNs. These results suggest that the delayed apoptosis in the brachial MN population is not a result of differing peripheral trophic support, but is intrinsically programmed, at least partially through a higher sensitivity to trophic factors.
The murine cripto gene: expression during mesoderm induction and early heart morphogenesis
Authors: Dono Rosanna , Scalera Lorenzo , Pacifico Francesco , Acampora Dario , Persico M. Graziella , Simeone Antonio 
Published: April 2021

ABSTRACT
The murine cripto gene encodes a 171-aminoacid epidermal growth factor-related protein, with 93% similarity to its human counterpart in the ‘EGF-like’ domain. The murine cripto mRNA contains two B1 repeats in its 3 non-coding region and a 163-nucleotide homology to the human mRNA.
The mouse cripto gene is expressed at low level in specific organs of the adult animal such as spleen, heart, lung and brain. In situ hybridization analysis during murine embryogenesis (day 6.2 to day 10.5) reveals a very restricted expression pattern. cripto transcripts are first detected in a few epiblastic cells at day 6.5. During gastrulation, the transcripts are expressed in the forming mesoderm and later during development cripto gene expression is restricted to the truncus arteriosus of the developing heart. This expression pattern suggests a role for cripto gene in the determination of the epiblastic cells that subsequently give rise to the mesoderm.
Postembryonic proliferation of neuroendocrine cells expressing adipokinetic hormone peptides in the corpora cardiaca of the locust
Authors: Kirschenbaum Sheila R. , O’Shea Michael 
Published: April 2021

ABSTRACT
Neuroendocrine glands that synthesize and secrete peptide hormones regulate the levels of these peptide messengers during development. In this article we describe a mechanism for regulating neuropeptide levels in the corpora cardiaca of the locust Schistocerca gregaria, a neuroendocrine gland structurally analogous to the vertebrate adenohypophysis. A set of five colocalized peptide hormones of the adipokinetic hormone family is synthesized in intrinsic neurosecretory cells in the corpora cardiaca. During postembryonic development there are progressive changes in the absolute and relative levels of these five peptide hormones. We show that the ability of the gland to increase peptide synthesis is due to a 100-fold increase in the number of cells which make up the gland. The gland grows by the addition of new cells derived from symmetrical division of undifferentiated precursor cells within the corpora cardiaca. We show, using double-label immunocytochemistry, that cells born in the glandular lobe mature into cells that express adipokinetic hormone peptides. The pattern of cell birth and peptide expression can account for the dramatic increase in postembryonic peptide levels.
Mutations affecting the cytoskeletal organization of syncytial Drosophila embryos
Authors: Sullivan William , Fogarty Patrick , Theurkauf William 
Published: April 2021

ABSTRACT
Cytoplasmic organization, nuclear migration, and nuclear division in the early syncytial Drosophila embryo are all modulated by the cytoskeleton. In an attempt to identify genes involved in cytoskeletal functions, we have examined a collection of maternal-effect lethal mutations induced by single P-element transposition for those that cause defects in nuclear movement, organization, or morphology during the syncytial embryonic divisions. We describe three mutations, grapes, scrambled, and nuclear-fallout, which define three previously uncharacterized genes. Females homozygous for these mutations produce embryos that exhibit extensive mitotic division errors only after the nuclei migrate to the surface. Analysis of the microfilament and microtubule organization in embryos derived from these newly identified mutations reveal disruptions in the cortical cytoskeleton. Each of the three mutations disrupts the actin-based pseudocleavage furrows and the cellularization furrows in a distinct fashion. In addition to identifying new genes involved in cytoskeletal organization, these mutations provide insights into cytoskeletal function during early Drosophila embryogenesis.
Staging of gastrulating mouse embryos by morphological landmarks in the dissecting microscope
Authors: Downs Karen M. , Davies Tim 
Published: April 2021

ABSTRACT
We describe and illustrate a set of morphological landmarks for classifying mouse embryos by gross morphology from before gastrulation to the beginning of organogenesis. These landmarks are visible at the resolution of the dissecting microscope in embryos that are intact except for reflexion of Reichert’s membrane. Adoption of these criteria should facilitate interpretation of both the expression patterns of genes and the consequences of experimental manipulation of embryos during early postimplantation development.
expanded: a gene involved in the control of cell proliferation in imaginal discs
Authors: Boedigheimer Michael , Laughon Allen 
Published: April 2021

ABSTRACT
The expanded gene was first identified by a spontaneous mutation that causes broad wings. We have identified an enhancer-trap insertion within expanded and used it to generate additional mutations, including one null allele. expanded is an essential gene, necessary for proper growth control of imaginal discs and, when mutant, causes either hyperplasia or degeneration depending on the disc. Wing overgrowth in expanded hypermorphs is limited to specific regions along the anterior-posterior and dorsal-ventral axis. expanded encodes a novel 1429 amino acid protein that is localized to the apical surface of disc cells and contains three potential SH3-binding sites. Together, these observations suggest that the Expanded protein engages in protein-protein interactions regulating cell proliferation in discs.
A novel H+ permeability dominating intracellular pH in the early mouse embryo
Authors: Baltz Jay M. , Biggers John D. , Lechene Claude 
Published: April 2021

ABSTRACT
Most cell types are relatively impermeant to H+ and are able to regulate their intracellular pH by means of plasma membrane proteins, which transport H+ or bicarbonate across the membrane in response to perturbations of intracellular pH. Mouse preimplantation embryos at the 2-cell stage, however, do not appear to possess specific pH-regulatory mechanisms for relieving acidosis. They are, instead, highly permeable to H+, so that the intracellular pH in the acid and neutral range is determined by the electrochemical equilibrium of H+ across the plasma membrane. When intracellular pH is perturbed, the rate of the ensuing H+ flux across the plasma membrane is determined by the H+ electro-chemical gradient: its dependence on external K+ con-centration indicates probable dependence on membrane potential and the rate depends on the H+ concentration gradient across the membrane. The large permeability at the 2-cell stage is absent or greatly diminished in the trophectoderm of blastocysts, but still present in the inner cell mass. Thus, the permeability to H+ appears to be developmentally regulated.