bluuebunny/crossref_metadata_2025 huggingface dataset:
The extracellular matrix of lip wounds in fetal, neonatal and adult mice
Authors: Whitby David J. , Ferguson Mark W. J.
Published: April 2021

Wound healing in the fetus occurs rapidly, by a regenerative process and without an inflammatory response, resulting in complete restitution of normal tissue function. By contrast, in the adult, wounds heal with scar formation, which may impair function and inhibit further growth. The cellular mechanisms underlying these differing forms of wound healing are unknown but the extracellular matrix (ECM), through its effects on cell function, may play a key role. We have studied the ECM in upper lip wounds of adult, neonatal and fetal mice at days 14, 16 and 18 of gestation. The spatial and temporal distribution of collagen types I, in, IV, V and VI, fibronectin, tenascin, laminin, chondroitin and heparan sulphates were examined immunohisto-chemically. Results from the fetal groups were essentially similar whilst there were distinct differences between fetus, neonate and adult. Fibronectin was present at the surface of the wound in all groups at 1 h post-wounding. Tenascin was also present at the wound surface but the time at which it was first present differed between fetus (1 h), neonate (12 h) and adult (24 h). The time of first appearance paralleled the rate of wound healing which was most rapid in the fetus and slowest in the adult. Tenascin inhibits the cell adhesion effect of fibronectin and during development the appearance of tenascin correlates with the initiation of cell migration. During wound healing the appearance of tenascin preceded cell migration and the rapid closure of fetal wounds may be due to the early appearance of tenascin in the wound. Collagen types I, HI, IV, V and VI were present in all three wound groups but the timing and pattern of collagen deposition differed, with restoration of the normal collagen pattern in the fetus and a scar pattern in the adult. This confirms that lack of scarring in fetal wounds is due to the organisation of collagen within the wound and not simply lack of collagen formation. The distribution of chondroitin sulphate differed between normal fetal and adult tissues and between fetal and adult wounds. Its presence in the fetal wound may alter collagen fibril formation. No inflammatory response was seen in the fetal wounds. The differences in the ECM of fetal and adult wounds suggests that it may be possible to alter the adult wound so that it heals by a fetal-like process without scar formation, loss of tissue function or restriction of growth.
Localized and inducible expression of Xenopus-posterior (Xpo), a novel gene active in early frog embryos, encoding a protein with a ‘CCHC’ finger domain
Authors: Sato Sheryl M. , Sargent Thomas D.
Published: April 2021

Xenopus-posterior (Xpo) is a gene that is activated at or shortly after the midblastula transition (MBT). The RNA accumulates to a relatively low level, which remains constant until gastrulation, then rapidly and transiently increases in posterior ectoderm and mesoderm. A single copy of a putative finger motif, of the ‘CCHC’ type, is located near the carboxyl terminus. One or two copies of similar sequence motifs are found in the nucleocapsid protein of retroviruses where they are involved in protein-RNA interactions, and in cellular nucleic acid binding protein (CNBP), a protein that binds to the sterol regulatory element. Xpo expression is induced in ectodermal expiants by treatment with basic fibroblast growth factor (bFGF) and with polypeptide growth factors found in medium conditioned by the Xenopus XTC cell line (XTC-CM). Taken together, these properties suggest a possible role for Xpo in the organization of the anteroposterior axis during development.
Gastrulation in Drosophila: the formation of the ventral furrow and posterior midgut invaginations
Authors: Sweeton Dari , Parks Suki , Costa Michael , Wieschaus Eric
Published: April 2021

The ventral furrow and posterior midgut invaginations bring mesodermal and endodermal precursor cells into the interior of the Drosophila embryo during gastrulation. Both invaginations proceed through a similar sequence of rapid cell shape changes, which include apical flattening, constriction of the apical diameter, cell elongation and subsequent shortening. Based on the time course of apical constriction in the ventral furrow and posterior midgut, we identify two phases in this process: first, a slow stochastic phase in which some individual cells begin to constrict and, second, a rapid phase in which the remaining unconstricted cells constrict. Mutations in the concertina or folded gastrulation genes appear to block the transition to the second phase in both the ventral furrow and the posterior midgut invaginations.
The L5 epitope: an early marker for neural induction in the chick embryo and its involvement in inductive interactions
Authors: Roberts Catherine , Platt Nicholas , Streit Andrea , Schachner Melitta , Stern Claudio D.
Published: April 2021

The pattern of expression of the carbohydrate epitope L5 was studied during early development of the chick neuroepithelium. Immunoreactivity first appears during gastrulation, at mid-primitive streak stage, and persists until at least 3.5 days of development. The epitope is expressed on all the components of the developing nervous system, both central and peripheral. In immunoblots, the antibody recognises a major component of about Mr 500000 and several more minor components of lower molecular mass.

If a Hensen’s node from a donor embryo is transplanted into the area opaca of a host embryo, L5 immunoreactivity appears in the epiblast surrounding the graft. If hybridoma cells secreting the antibody are grafted together with Hensen’s node into a host chick embryo, the induction of a supernumerary nervous system is inhibited. We suggest that the L5 epitope is an early and general marker for neural induction and that it may be involved directly in inductive interactions.
Pleiotropic effect of okadaic acid on maturing mouse oocytes
Authors: Alexandre H. , Van Cauwenberge A. , Tsukitani Y. , Mulnard J.
Published: April 2021

Okadaic acid (OA), a potent inhibitor of types 1 and 2A protein phosphatases, was shown recently to induce chromatin condensation and germinal vesicle breakdown (GVBD) in mouse oocytes arrested at the dictyate stage by dibutyryl cAMP (dbcAMP), isobutyl methylxanthine (IBMX) and 12,13-phorbol dibutyrate (PDBu). We confirm these results using IBMX and another phorbol diester, 12-O-tetradecanoylphorbol-13-acetate (TPA) and show that OA also bypasses the inhibitory effect of 6-dimethylaminopurine (6-DMAP). It has been concluded that protein phosphatases 1 and/or 2A (PP1, 2A), involved in the negative control of MPF activation, are thus operating downstream from both the protein kinase A and protein kinase C catalysed phosphorylation steps that prevent the breakdown of GV. Similar enzymatic activities are also able to counteract the general inhibition of protein phosphorylation.

However, PP1 and/or PP2A are positively involved in the activation of pericentriolar material (PCM) into microtubule organizing centres (MTOCs). This explains the inhibitory effect of OA on spindle assembly.

Finally, OA interferes with the integrity and/or function of actomyosin filaments. This results in a dramatic ruffling of the plasma membrane leading to the internalization of large vacuoles, the inhibition of chromosome centrifugal displacement and, consequently, the prevention of polar body extrusion.
Patterns of E74A RNA and protein expression at the onset of metamorphosis in Drosophila
Authors: Boyd Lynn , O’Toole Erin , Thummel Carl S.
Published: April 2021

Metamorphosis in Drosophila is triggered by a pulse of the steroid hormone ecdysone at the end of larval development. Ecdysone initiates a genetic hierarchy that can be visualized as a series of puffs in the larval salivary gland polytene chromosomes. The E74 gene is responsible for the early ecdysone-inducible puff at position 74EF and encodes two related DNA-binding proteins which appear to play a regulatory role in the hierarchy. Here we describe the spatial and temporal patterns of E74A RNA and protein expression at the onset of metamorphosis. We use in situ hybridization, antibody stains, and western and northern blot analyses to follow E74A expression from its initial appearance as nascent transcripts on the polytene chromosomes, to spliced mRNA, to post-translationally modified nuclear E74A protein. E74A is expressed in a wide variety of late-third instar tissues, suggesting that it plays a broad pleiotropic role in response to the hormone. In early prepupae, when the overall levels of E74A mRNA are decreasing, relatively high levels of E74A RNA persist in the gut, peripodial membranes of the imaginai discs, and proliferation centers of the brain. The spatial distribution of nuclear E74A protein correlates with the RNA distribution with the single exception that no E74A protein can be detected in the proliferation centers of the brain. There is also a temporal discrepancy between E74A mRNA and protein accumulation. The peak of E74A protein induced by the late larval ecdysone pulse follows the peak of E74A mRNA by ∼2h. This delay is not seen in 10 h prepupae, when the next pulse of ecdysone induces the simultaneous expression of E74A mRNA and protein. We discuss possible mechanisms for post-transcriptional regulation of E74A expression and suggest that the unusually long and complex 5′ leader in the E74A mRNA may regulate its translation.
‘Promoter trapping’ in Caenorhabditis elegans
Authors: Hope I. A.
Published: April 2021

A screen of gene expression patterns has been developed for the nematode Caenorhabditis elegans. Promoterreporter gene fusions were constructed in vitro by ligating C.elegans genomic DNA fragments upstream of a lacZ gene. Patterns of β-galactosidase expression were examined by histochemical staining of C. elegans lines transformed with the constructs, β-galactosidase expression depended on translational fusion, so constructs were assayed in large pools to expedite detection of the low proportion that were active. Expression in a variety of cell types and temporal patterns was observed with different construct pools. The most striking expression patterns were obtained when the β-galactosidase activity was localized to subcellular structures by the C. elegans portion of the fusion protein. The active constructs of three selected pools were identified subsequently by an efficient combinatorial procedure. The genomic locations of the DNA fragments from the active constructs were determined and appear to define previously uncharacterized genetic loci.
Fate maps of the first quartet micromeres in the gastropod llyanassa obsoleta
Authors: Render Joann
Published: April 2021

Cell fate specification in the gastropod mollusc Ilyanassa obsoleta involves both cell autonomous and inductive mechanisms, which depend on determinants localized first in the polar lobe and then in the D quadrant of the embryo. A complete cell lineage is lacking for this embryo and is essential for a critical interpretation of previous experimental results and an analysis of the mechanisms at the molecular level.

Lineages of the first quartet micromeres were followed using Lucifer Yellow dextran as a tracer. The tracer was injected into individual first quartet micromeres using iontophoresis and patterns of fluorescence were analyzed in the larva after 8 days of development. Fluorescence was limited to head structures, including eyes, tentacles and velum. Structures on the left side were derived from 1a and 1d micromeres; la gave rise to the left eye, including the lens. Right side structures were derived from the 1c micromere and 1b contributed to the apical plate between the eyes and symmetrically to both sides of the velum. First quartet lineage data are compared with results from previous cell ablation experiments and with lineage data from other species.
Production of antisense RNA leads to effective and specific inhibition of gene expression in C. elegans muscle
Authors: Fire Andrew , Albertson Donna , Harrison Susan White , Moerman Donald G.
Published: April 2021

We have used an antisense strategy to effectively disrupt the expression of two genes encoding myofilament proteins present in C. elegans body wall muscles. DNA segments from the unc-22 and unc-54 genes have been placed in reverse orientation in vectors designed to produce RNA in body wall muscles. When the resulting plasmids are injected into oocytes, progeny with defects in muscle function are produced. These animals have phenotypes consistent with reduction and/or elimination of function of the gene to which antisense RNA has been produced: twitching and disorganization of muscle filaments for the unc-22 antisense constructs and lack of muscle tone, slow movement, and egg laying defects for the unc-54 antisense constructs. A fraction of the affected animals transmit the defective-muscle trait to subsequent generations. In these cases the transforming DNA is present at high copy number and cosegregates with the observed muscle defects. We have examined several of the unc-22 antisense plasmid transformed lines to determine the mechanistic basis for the observed phenotypes. The RNA product of the endogenous unc-22 locus is present at normal levels and this RNA is properly spliced in the region homologous to the antisense RNA. No evidence for modification of this RNA by deamination of adenosine to inosine was found. In affected animals the level of protein product from the endogenous unc-22 locus is greatly reduced. Antisense RNA produced from the transforming DNA was detected and was much more abundant than ‘sense’ RNA from the endogenous locus. These data suggest that the observed phenotypes result from interference with a late step in gene expression, such as transport into the cytoplasm or translation.
Binding of a factor to an enhancer element responsible for the tissue specific expression of the chicken αA-crystallin gene
Authors: Matsuo Isao , Kitamura Masahi , Okazaki Kenji , Yasuda Kunio
Published: April 2021

We have characterized a regulatory region of the chicken αA-crystallin gene using transfection assays, which revealed that a 84 base pair element (–162 to –79) in the 5’ flanking sequence is necessary and sufficient for lens-specific expression. A multimer of this element functions as lens-specific enhancer and synergistically activates transcription from chicken αA-crystallin or β-actin basal promoters fused to the CAT gene. In vivo competition experiments demonstrated that DNA sequences containing the 84 bp element reduced αA-crystallin–CAT fusion gene expression. A nuclear factor present exclusively in lens cells binds to the 84 bp element in the region between positions –165 and –140. Southwestern blot analysis showed that 61000 Mr (61×103Mr) lens nuclear protein exhibited DNA-binding activity specific to the 84 bp element. Our data suggested that the 61×103Mr nuclear protein, and the 84 bp element that it interacts with, may be involved in regulating the αA-crystallin gene expression in vivo.
Generation of pigmented stripes in albino mice by retroviral marking of neural crest melanoblasts
Authors: Huszar Dennis , Sharpe Arlene , Hashmi Sofie , Bouchard Brigitte , Houghton Alan , Jaenisch Rudolf
Published: April 2021

The pigment cells of the skin are derived from melanoblasts which originate in the neural crest. The dorsoventral migration of melanoblasts has been visualized in pigment stripes seen in aggregation chimeras, and the width of these bands has suggested that the entire pigmentation of the coat is derived from a small number of founder cells. We have generated mosaic mice by marking single melanoblasts in utero to gain information on the clonal history of pigment-forming cells. A retroviral vector carrying the human tyrosinase gene was constructed and microinjected into neurulating albino mouse embryos. Albino mice are devoid of pigmentation due to deficiency of tyrosinase. Thus, transduction of the wild-type gene into the otherwise normal melanoblasts should rescue the mutant phenotype, giving rise to patches of pigmentation, which correspond to the area colonized by the mitotic progeny of a marked clone. Mosaic animals derived from the injected embryos indeed showed pigmented bands with a width strikingly similar to the ‘standard’ stripes seen in aggregation chimeras. These results are consistent with the notion that the unit width bands seen in aggregation chimeras represent the clonal progeny of a single melanoblast and verify Mintz’s (1967) conclusion that a few founder melanoblasts give rise to coat pigmentation. The pigment cells of the eye are of dual origin: the melanocytes in choroid and outer layer of the iris are derived from the neural crest and those in the pigment layer of the retina from the neuroepithelium of the optic cup. Marked clones in both lineages were observed in the eyes of many mosaic animals.
The nature of the mesoderm-inducing signal in Xenopus: a transfilter induction study
Authors: Slack J. M. W.
Published: April 2021

A transfilter apparatus is described, which is suitable for neutralization experiments on embryonic induction, and it is used to investigate the sensitivity of the Xenopus mesoderm-inducing signal to various inhibitors. The vegetal (inducing) tissue is placed on one side of a membrane sandwich and the animal (responding) tissue on the other side. The sandwich consists of a nylon gauze in between two Nucleopore filters and enables inhibitors in the solution to have effective access to the gap between the tissues. Control experiments show a high proportion of positive inductions of a ventral character.

Using this apparatus, it is shown that the protein follistatin, which effectively inhibits activin A and B in vitro, has little or no effect on the natural signal. Likewise, antibodies to basic fibroblast growth factor, which inhibit in vitro, do not inhibit the natural signal. The two inhibitors together have a slight effect. It is concluded that neither activin nor bFGF are major components of the signal emitted by the vegetal cells of the Xenopus blastula and transmitted across the liquid gap, although they might have some other role to play in the process.

Two agents of lower specificity do inhibit the transfilter induction: heparin and suramin. Suramin will also inhibit induction in animal-vegetal combinations with no intervening membranes while heparin does not. This suggests that the heparin inhibition can only occur when there is a liquid gap between the tissues, presumably because it can neutralize the signal in solution but cannot penetrate the explants themselves. The endogenous mesoderm-inducing factor(s) should therefore be sensitive to heparin in vitro.
The Mus musculus domesticus Tdy allele acts later than the Mus musculus musculus Tdy allele: a basis for XY sex-reversal in C57BL/6-YPOS mice
Authors: Palmer Stephen J. , Burgoyne Paul S.
Published: April 2021

Consomic C57BL/6 males, carrying either the Mus musculus musculus-derived C57BL/6 Y chromosome or the Mus musculus domesticus-derived Poschiavinus Y chromosome, were outcrossed to females of the inbred strains C3H/Bi and CXBH/By and to females of the random bred strain MF1/Ola. In a study at 12.5 days post coitum, gonads of XYCS7 and XYPOS fetuses were assessed for the presence of testicular cords. It was found that XYPOS fetuses had a later onset of testicular development than XY57 fetuses. Limb development, which was monitored as a measure of overall development, was unaffected by the strain of Y present. These data were supported by a longitudinal study in which the increased growth rate of the testes relative to undifferentiated gonads, was also shown to be delayed in XYPOS fetuses. The extent of the delay was estimated to be approximately 14 h. It is concluded that this delay in the onset of testicular differentiation must be caused by differences between the two Y-chromosome types, most probably allelic differences in the testis determinant Tdy.
Developmental analysis of the eye lens obsolescence (Elo) gene in the mouse: cell proliferation and Elo gene expression in the aggregation chimera
Authors: Yoshiki Atsushi , Hanazono Makoto , Oda Sen-Ichi , Wakasugi Noboru , Sakakura Teruyo , Kusakabe Moriaki
Published: April 2021

This study investigates the primary effect of the eye lens obsolescence (Elo) gene of the mouse. Morphological features of the Elo lens were defined as follows: (1) deficient elongation of lens fiber cells, (2) morphological abnormality of nuclei of lens fiber cells, (3) lack of eosinophilic granules in the central fiber cells and (4) rupture of lens capsule in the posterior region. We have immunohistologically examined, by means of an in vivo BrdU incorporation system, whether or not the Elo gene regulates cell proliferation during lens development. The lens fiber cells were morphologically abnormal in day 13 embryonic Elo lens. However, there were no significant differences in morphology or cell proliferation between normal and Elo lens epithelium until day 14 of gestation. After day 15, the total cell number in the Elo lens epithelium was significantly less than that in the normal, but the total numbers of S-phase cells in the two genotypes were not significantly different. The ratio of the total S-phase cell number to the total number of lens epithelial cells may be affected by the developmental stage, but not directly by the genotype. The genotype, however, may be having a direct influence at later ages because malformation of Elo lens fiber cells must cause reduction of the total number of lens epithelial cells in older embryos. Although, at 30 days old, Elo lens cells were externally extruded through the ruptured capsule into the vitreous cavity, BrdU-labelled lens epithelial cells were detectable. To investigate whether the Elo lens phenotype is determined by its own genotype or by its cellular environment, we produced aggregation chimeras between C3H-Elo/+(C/C) and BALB/c (c/c). Most lenses of BALB/c dominant chimeras were oval in shape without the ruptured lens capsule. However, they were opaque in the center and slightly smaller in size than normal. The lenses of C3H-E/o/ + dominant chimeras were morphologically similar to the Elo lens. Although normal nuclei were regularly arranged in the anterior region, Elo-type nuclei were located in the posterior region. Immunohistological staining by using anti-C3H strain-specific antibody demonstrated that the lens fiber cells with abnormal nuclei were derived only from C3H-Elo/+, not from BALB/c. These observations suggest that the primary effect of the Elo gene in the developing lens may be specific to the fiber cell differentiation rather than to the cell proliferation. Moreover, the Elo gene may be autonomously expressed in the differentiating lens fiber cells, and intracellularly inhibit fiber cell elongation.
Evidence that hypophysis and hypothalamus constitute a single entity from the primary stage of histogenesis
Authors: Kawamura Kosuke , Kikuyama Sakae
Published: April 2021

To identify the origin of the primordial pituitary cells in early stages of amphibian embryos before the formation of Rathke’s pouch or its equivalent, transplantation experiments were carried out on Bufo. Wild-type embryos at the open neurula stage were employed as donors and albino embryos at the corresponding stage as recipients. Since brain and pituitary cells of wild-type animalscontain a great number of melanin granules, whereas albino cells are devoid of them, the melanin granules enabled the fate of transplants to be traced.

The anterior part of the neural ridge (ANR) was found to be the almost exclusive source of the secretory cells in the epithelial pituitary gland (pars distalis, pars intermedia and pars tuberalis). Transplantation of either the central part of the anterior neural plate (NP) or the ANR revealed that the pituitary primordium and the anlage of the posterior hypothalamus (infundibulum) are closely apposed at the open neurula stage. As morphogenetic movement proceeds, each of the two entities takes a separate migration route. The former part (NP) takes a ventrocaudal route to become the diencephalic floor, and the latter part (ANR) takes a rostroventral route to acquire morphological contact with the anterodorsal wall of the foregut before differentiating into the epithelial hypophysis. At the end of the tail-bud stage, they meet again beneath the diencephalic floor and establish a hypothalamo-hypophyseal connection. Some of the ANR cells are incorporated into the preoptic region of the anterior hypothalamus. None of the NP cells develop into epithelial pituitary cells.

We conclude that the epithelial pituitary gland is not stomodeal, but placodal, in origin and that the hypothalamo-hypophyseal complex may be regarded as a single entity from an ontogenetic viewpoint.
Developmentally regulated expression ofα6 integrin in avian embryos
Authors: Bronner-Fraser Marianne , Artinger Michael , Muschler Iohn , Horwitz Alan F.
Published: April 2021

The distribution pattern of the avian α6 integrin subunit was examined during early stages of development. The results show that this subunit is prevalent in cells of the developing nervous system and muscle. α6 is first observed on neuroepithelial cells of the cranial neural plate and trunk neural tube. With time, immunoreactivity becomes prominent near the lumen and ventrolateral portions of the neural tube, co-distributing with neurons and axons, particularly notable on commissural neurons. The α6 expression pattern is dynamic in the nenral tube, with immunoreactivity peaking by embryonic day 6 (stage 30) and decreasing thereafter. The ventral roots and retina exhibit high levels of immnno-reactivity throughout development. In the peripheral nervous system, α6 immunoreactivity first appears on a subpopulation of sympathoadrenal cells around the dorsal aorta and later in the dorsal root ganglia shortly after gangliogenesis. Immunoreactivity appears on pro-spective myotomal cells as the somites delaminate into the dermomyotome and sclerotome, remaining prominent on myoblasts and differentiated muscle at all stages. The mesonephros also has intense immunoreactivity. In the periphery, α6 immunoreactive regions often in proximity to laminin, which is thought to be the ligand of α6β1 integrin.
Developmental expression of the α receptor for platelet-derived growth factor, which is deleted in the embryonic lethal Patch mutation
Authors: Orr-Urtreger Avi , Bedford Mark T. , Do Myoung-Sool , Eisenbach Lea , Lonai Peter
Published: April 2021

The α receptor of PDGF (Pdgfra) is expressed in primitive endoderm and mesoderm derivatives through-out embryogenesis. In the early primitive streak stage the gene is transcribed in the visceral and parietal endoderm. Later it is expressed in the presomitic mesoderm, yolk sac and amnion. During somitogenesis its transcription localizes to the heart and the somites. Subsequently, it is transcribed in the dermatome, the sclerotome, the developing limb and in various mesenchymal tissues of visceral organs. Its wild-type expression pattern correlates well with the phenotype of homozygous mutant Patch (Ph) embryos, where the Pdgfra gene is deleted. The Ph phenotype is first detectable at the primitive streak stage with convoluted and hypertrophic visceral yolk sac, deformed neural plate and disorganized or missing mesoderm. Most Ph/Ph embryos die before the 11th day of gestation. Those that survive till early organogenesis are very small, have hypertrophic yolk sacs, small and undifferentiated somites, convoluted neural tubes, large heart and pericardium, rudimentary limb buds and branchial arches. Our observations together suggest that the α PDGF receptor may be required for the normal development of visceral endoderm and mesoderm derivatives.
A unique mutation in the Enhancer of split gene complex affects the fates of the mystery cells in the developing Drosophila eye
Authors: Fischer-Vize Janice A. , Vize Peter D. , Rubin Gerald M.
Published: April 2021

An unusual recessive allele of the Drosophila groucho gene, which encodes a transducinlike protein, affects the fates of specific cells in the eye disc groucho is one of several transcription units in the Enhancer of split complex. Most groucho mutations are zygotic lethal due to the proliferation of embryonic neural cells at the expense of epidermal cells. In contrast, flies homozygous for the mutant allele described here, groBFP2, are viable but have abnormal eyes. The Drosophila compound eye is composed of several hundred identical facets, or ommatidia, each of which contains eight photoreceptor cells, R1-R8. In groBFP2 mutant retinas, most of the facets contain eight normally determined photoreceptor cells and one or two additional R-cells of the R3/4 subtype. The extra photoreceptors appear to arise from the mystery cells, which are part of the precluster that initiates the ommatidium, but do not normally become neurons. groBFP2 behaves as a partial loss-of-function mutant. Analysis of ommatidia mosaic for wild-type and groBFP2 mutant cells suggests that the focus of action of the groBFP2 mutation is outside of the photoreceptor cells. These results imply that one function of groucho is in a pathway whereby neuralization of the mystery cells is inhibited by other non-neural cells in the eye disc. In addition, determination of R3/4 photoreceptors usually requires contact with R2 and R5. Specification of the mystery cells as ectopic R3/4 subtype photoreceptors in groBFP2 mutant eye discs implies that induction by R2 or R5 is not absolutely necessary for R3/4 cell determination.
Respecification of vertebral identities by retinoic acid
Authors: Kessel Michael
Published: April 2021

In higher vertebrates, the formation of the body axis proceeds in a craniocaudal direction during gastrulation. Cell biological evidence suggests that mesoderm formation and specification of axial positions occur simultaneously. Exposure of gastrulating embryos to retinoic acid induces changes in axial patterns, e.g. anterior and posterior homeotic transformations of vertebrae. These morphological changes are accompanied by changes in the nonidentical, overlapping expression domains of Hox genes.

In this report the influence of retinoic acid, administered at the end of and after gastrulation, on vertebral patterns is described. Anterior transformations and truncations affecting the caudal part of the vertebral column characterize animals exposed on day 8 and 9. 4 hours after retinoic acid administration on day 8 + 5 hours, Hox-1.8, Hox-1.9, and Hox-4.5 transcripts were not detected in their usual posterior expression domains, whereas transcripts of the anterior Hox-1.5 gene remained unaffected. 4 days after RA exposure on day 8 + 5 hours, Hox-1.8 expression was shifted posteriorly by an effectively low dose of RA, which induced the formation of supernumerary ribs. Hox-1.8 expression was limited to posterior, disorganized mesenchyme, bulging out neural tube, some intestinal loops and the hindlimb in truncated embryos exposed to a high dose of RA. A causal relation between the delayed activation of posterior Hox genes and anterior transformations or agenesis of vertebrae is discussed.

On day 10.5 posterior transformations begin to occur in the cervical region, while later exposures again affect more caudal structures. The distribution of the transformations along the vertebral column indicates an influence of RA on migrating sclerotome cells before they are finally fixed in the cartilagenous vertebrae. The findings show that the mesodermal segments originally specified during gastrulation can be respecified in their second migratory phase, with effects spreading for a second time in a craniocaudal direction. The transformations are discussed with regard to a molecular specification of axial levels by Hox codes, defined as combinations of expressed Hox genes.
ventral ectoderm of Xenopus forms neural tissue, including hindbrain, in response to activin
Authors: Bolce1 Margarete. , Hemmati-Brivanlou Ali , Kushner Pinky D. , Harland Richard M.
Published: April 2021

The peptide growth factor Activin A has been shown to induce complete axial structures in explanted blastula animal caps. However, it is not understood how much this response to activin depends upon early signals that prepattern the ectoderm. We have therefore asked what tissues can be induced in blastula animal caps by activin in the absence of early dorsal signals. Using whole mount in situ hybridization, we compare the expression of three neural markers, N-CAM, En-2 and Krox-20 in activin-treated ectoderm from control and ventralized embryos. In response to activin, both normal and ventralized animal caps frequently form neural tissue (and express N-CAM) and express the hindbrain marker Krox-20. However, the more anterior marker, En-2, is expressed in only a small fraction of normal animal caps and rarely in ventralized animal caps; the frequency of expression does not increase with higher doses of activin. In all cases En-2 and Krox-20 are expressed in coherent patches or stripes in the induced caps. Although mesoderm is induced in both control and ventralized animal caps, notochord is found in response to activin at moderate frequency in control caps, but rarely in ventralized animal caps. These results support the idea that in the absence of other signals, activin treatment elicits hindbrain but not notochord or anterior neural tissue; and thus, the anterior and dorsal extent of tissues formed in response to activin depends on a prior prepatterning or previous inductions.
A fate map of the Arabidopsis embryonic shoot apical meristem
Authors: Irish Vivian F. , Sussex Ian M.
Published: April 2021

We have mapped the fate of cells in the Arabidopsis embryonic shoot apical meristem by irradiating seed and scoring the resulting clonally derived sectors. 176 white, yellow, pale green or variegated sectors were identified and scored for their position and extent in the resulting plants. Most sectors were confined to a fraction of a leaf, and only occasionally extended into the inflorescence. Sectors that extended into the inflorescence were larger, and usually encompassed about a third to a half of the inflorescence circumference. We also find that axillary buds in Arabidopsis are clonally related to the subtending leaf. Sections through the dry seed embryo indicate that the embryonic shoot apical meristem contains approximately 110 cells in the three meristematic layers prior to the formation of the first two leaf primordia. The histological analysis of cell number in the shoot apical meristem, in combination with the sector analysis have been used to construct a map of the probable fate of cells in the embryonic shoot apical meristem.
Contrasting patterns of c-myc and N-myc expression in proliferating, quiescent, and differentiating cells of the embryonic chicken lens
Authors: Harris Laura L. , Talian John C. , Zelenka Peggy S.
Published: April 2021

The present study uses the polymerase chain reaction and in situ hybridization to examine c-myc and N-myc mRNA in the embryonic chicken lens at 6,10,14 and 19 days of development and compares the pattern of expression obtained with the developmental pattern of cell proliferation and differentiation. In the central epithelium, c-myc mRNA levels were proportional to the percentage of proliferating cells throughout development. N-myc mRNA expression in this region was relatively low and showed no correlation with cell proliferation. The ratio of N-myc to c-myc mRNA increased markedly with the onset of epithelial cell elongation and terminal fiber cell differentiation, although both c-myc and N-myc mRNAs continued to be expressed in postmitotic, elongating cells of the equatorial epithelium and in terminally differentiating lens fiber cells. Thus, increased expression of N-myc, a gene whose protein product may compete with c-myc protein for dimerization partners, accompanies the dissociation of c-myc expression and cell proliferation during terminal differentiation of lens fiber cells.
Molecular organization and embryonic expression of the hedgehog gene involved in cell-cell communication in segmental patterning of Drosophila
Authors: Mohler Jym , Vani Kodela
Published: April 2021

hedgehog is a segment polarity gene necessary to maintain the proper organization of each segment of the Drosophila embryo. We have identified the physical location of a number of rearrangement breakpoints associated with hedgehog mutations. The corresponding hh RNA is expressed in a series of segmental stripes starting at cellular blastoderm in the posterior portion of each segment. This RNA is localized predominantly within nuclei until stage 10, when the localization becomes primarily cytoplasmic. Expression of hh RNA in the posterior compartment is independent of most other segment polarity genes, including en, until the late extended germ-band stage (stage 11). Sequence analysis of the hedgehog locus suggests the protein product is a transmembrane protein, which may, therefore, be directly involved in cell-cell communication.
3′ non-translated sequences in Drosophila cyclin B transcripts direct posterior pole accumulation late in oogenesis and peri-nuclear association in syncytial embryos
Authors: Dalby Brian , Glover David M.
Published: April 2021

We have characterised forms of the Drosophila cyclin B transcript that differ as a result of a splicing event which removes a nucleotide segment from the 3 untranslated region. In oogenesis, both cyclin A RNA and a shorter form of the cyclin B transcript are seen in the cells of the germarium that are undergoing mitosis. The shorter cyclin B transcript alone is then detectable in the presumptive oocyte until stages 7-8 of oogenesis. Both cyclin A RNA and a longer form of the cyclin B RNA are then synthesised in the nurse cells during stages 9-11, to be deposited in the oocyte during stages 11-12. These transcripts become evenly distributed throughout the oocyte cytoplasm but, in addition, those of cyclin B become concentrated at the posterior pole. Examination of the distributions of RNAs transcribed from chimeric cyclin genes indicates that sequences in the 3 untranslated region of the larger cyclin B RNA are required both for it to become concentrated at the posterior pole and to direct those transcripts in the body of the syncytial embryo to their peri-nuclear localisation. These sequences are disrupted by the splicing event which generates smaller cyclin B transcripts.
Expression of en and wg in the embryonic head and brain of Drosophila indicates a refolded band of seven segment remnants
Authors: Schmidt-Ott Urs , Technau Gerhard M.
Published: April 2021

Based on the expression pattern of the segment polarity genes engrailed and wingless during the embryonic development of the larval head, we found evidence that the head of Drosophila consists of remnants of seven segments (4 pregnathal and 3 gnathal) all of which contribute cells to neuromeres in the central nervous system. Until completion of germ band retraction, the four pregnathal segment remnants and their corresponding neuromeres become arranged in an S-shape. We discuss published evidence for seven head segments and morphogenetic movements during head formation in various insects (and crustaceans).
Expression of tenascin mRNA in mesoderm during Xenopus lævis embryogenesis: the potential role of mesoderm patterning in tenascin regionalization
Authors: Umbhauer Muriel , Riou Jean-François , Spring Jürg , Smith James C. , Boucaut Jean-Claude
Published: April 2021

In Xenopus embryos, the extracellular matrix (ECM) protein tenascin (TN) is expressed dorsally in a very restricted pattern. We have studied the spatial and temporal expression of TN mRNA in tailbud-stage embryos by RNAase protection and in situ hybridization using a cDNA probe for Xenopus TN obtained by PCR amplification. We report that TN transcripts are principally expressed in cells dispersed around the neural tube and notochord as well as in myotome and sclerotome cells. No TN mRNA could be detected in lateral plate mesoderm, but expression was detectable beneath tail fin epidermis. In a second series of experiments, we studied the expression of TN mRNA and protein in combinations between animal and vegetal stage-6 blastomeres and in stage-8 blastula animal caps treated with activin A or basic fibroblastic growth factor (b-FGF). Isolated animal cap tissue cultured alone differentiates into epidermis, which expresses neither TN protein nor TN mRNA. TN expression is, however, elicited in response to isolated dorsal vegetal blastomeres and in response to high concentrations of activin, both of which treatments lead to formation of muscle and/or notochord. Low concentrations of activin, and ventral vegetal blastomeres, treatments that induce mesoderm of ventral character, are poor inducers of TN. However, b-FGF, which also induces ventral mesoderm, elicits strong expression. These results indicate that TN regionalization is a complex process, dependent both on the pattern of differentiation of mesodermal tissues and on the agent with which they are induced. The data further show that “ventral mesoderm” induced by low concentrations of activin is distinct from that induced by b-FGF, and imply that activin induces ventral mesoderm of the trunk while b-FGF induces posterior mesoderm of the tailbud.
A dominant mutation in the maize homeobox gene, Knotted-1, causes its ectopic expression in leaf cells with altered fates
Authors: Smith Laurie G. , Greene Ben , Veit Bruce , Hake Sarah
Published: April 2021

Dominant mutations of the Knotted-1 (Kn1) homeobox gene of maize alter the differentiation and growth of cells associated with leaf veins. By analyzing Kn1 transcripts and KN1 protein, we show that the gene is not expressed at high levels during the development of wildtype leaves. Instead, Kn1 is expressed in apical meristems of vegetative and floral shoots, and is downregulated as leaves and floral organs are initiated. Kn1 is also expressed in relatively undifferentiated cells within developing vascular bundles, as well as ground tissue, in immature, unelongated axes of wild-type vegetative and floral shoots. In Kn1-N2 mutant plants, quantitative, but not qualitative differences are apparent in Kn1 transcripts and KN1 protein, consistent with previous observations that dominant Kn1 mutations map to noncoding regions of the gene. Kn1 is expressed ectopically in vascular bundles within developing mutant leaves in a pattern that correlates with the phenotypic alterations produced by the Kn1-N2 mutation. Thus, Kn1 apparently alters the fates of leaf cells in which it is ectopically expressed from an early stage of leaf development. Based on these observations, we hypothesize that Kn1 functions in its wild-type context as a regulator of cell determination.
Cloning and developmental expression of Sna, a murine homologue of the Drosophila snail gene
Authors: Nieto M. Angela , Bennett Michael F. , Sargent Michael G. , Wilkinson David G.
Published: April 2021

The genetic analysis of dorsoventral patterning in Drosophila has identified a zinc-finger gene, snail, that is required for mesoderm formation. The cloning and nuclease protection analysis of a Xenopus homologue of this gene has suggested a possible role in the mesoderm of vertebrates. Here, we describe the cloning of a murine homologue of snail, Sna, and in situ hybridisation studies of its developmental expression. Sequence analysis reveals substantial conservation of the second to fifth zinc fingers, but not of the first zinc finger in the Sna gene. Expression occurs in the ectoplacental cone, parietal endoderm, embryonic and extraembryonic meso-derm, in neural crest and in condensing precartilage. Based on the timing and spatial restriction of expression in embryonic mesoderm, we suggest that Sna might be required for the early development of this tissue, as is the case for its Drosophila counterpart. In addition, we propose that Sna might have an analogous role in the development of neural crest. The expression in condensing precartilage indicates that this gene also has a later function in chondrogenesis.
Transforming growth factor-β control of cell-substratum adhesion during avian neural crest cell emigration in vitro
Authors: Delannet Muriel , Duband Jean-Loup
Published: April 2021

It has been proposed that, in higher vertebrates, the onset of neural crest cell migration from the neural tube involves spatially and temporally coordinated changes in cellular adhesiveness that are under the control of external signals released in the extracellular milieu by neighboring tissues. In the present study, we have analyzed the dynamics of changes in cell-substratum adhesiveness during crest cell emigration and searched for regulatory cues using an in vitro model system. This model is based on the fact that, in vivo, crest cell dispersion occurs gradually along a rostrocaudal wave, allowing us to explant portions of the neural axis, termed migratory and premigratory levels, that differ in the time in culture at which neural crest cells initiate migration and in the locomotory behavior of the cells. We found that neural crest cell emigration is not triggered by the main extracellular matrix molecules present in the migratory pathways, as none of these molecules could abolish the intrinsic difference in the timing of emigration between the different axial levels. Using an in vitro adhesion assay, we found that presumptive neural crest cells from premigratory level explants gradually acquired the ability to respond to extracellular matrix material with time in culture, suggesting that acquisition of appropriate, functional integrin receptors was a necessary step for migration. Finally, we showed that members of the transforming growth factor-β family reduced in a dose-dependent manner the delay of neural crest cell emigration from premigratory level explants and were able to increase significantly the substratum-adhesion properties of crest cells. Our results suggest that acquisition of substratum adhesion by presumptive neural crest cells is a key event during their dispersion from the neural tube in vitro, and that members of the transforming growth factor-β family may act as potent inducers of crest cell emigration, possibly by increasing the substratum adhesion of the cells.
Synapse-specific expression of acetylcholine receptor genes and their products at original synaptic sites in rat soleus muscle fibres regenerating in the absence of innervation
Authors: Brenner H. R. , Herczeg A. , Slater C. R.
Published: April 2021

To test the hypothesis that synaptic basal lamina can induce synapse-specific expression of acetylcholine receptor (AChR) genes, we examined the levels mRNA for the and -subunits of the AChR in regenerating rat soleus muscles up to 17 days of regeneration. Following destruction of all muscle fibres and their nuclei by exposure to venom of the Australian tiger snake, new fibres regenerated within the original basal lamina sheaths. Northern blots showed that original mRNA was lost during degeneration. Early in regeneration, both and -subunit mRNAs were present throughout the muscle fibres but in situ hybridization showed them to be concentrated primarily at original synaptic sites, even when the nerve was absent during regeneration. A sim-ilar concentration was seen in denervated regenerating muscles kept active by electrical stimulation and in muscles frozen 41–44 hours after venom injection to destroy all cells in the synaptic region of the muscle. Acetylcholine-gated ion channels with properties similar to those at normal neuromuscular junctions were concentrated at original synaptic sites on denervated stimulated muscles. Taken together, these findings provide strong evidence that factors that induce the synapse-specific expression of AChR genes are stably bound to synaptic basal lamina.