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At least 631 records · Page 35Linked to original sources

PubMed1990

Early tissue interactions leading to embryonic lens formation in Xenopus laevis.

Our previous research has demonstrated that lens induction in Xenopus laevis requires inductive interactions prior to contact with the optic vesicle, which classically had been thought to be the major lens inductor. The importance of these early interactions has been verified by demonstrating that lens ectoderm is specified by the time it comes into contact with the optic vesicle. It has been argued that the tissues which underlie the presumptive lens ectoderm during gastrulation and neurulation, dorsolateral endoderm and mesoderm, are the primary early inductors. We show here, however, that these tissues alone cannot elicit lens formation in Xenopus ectoderm. Evidence is presented that presumptive anterior neural plate tissue (which includes the early eye rudiment) is an essential early lens inductor in Xenopus. The presence of dorsolateral mesoderm appears to enhance this response. These findings support a model in which an essential inductive signal passes through the plane of ectoderm during gastrula and early neurula stages from presumptive anterior neural tissue to the presumptive lens ectoderm. Since there is evidence for such interactions within a tissue layer in mesodermal and neural induction as well, this may be a general feature of the initial stages of determination of many tissues.

Animals↗

PubMed1990

Interspecific hybridization between an anural and urodele ascidian: differential expression of urodele features suggests multiple mechanisms control anural development.

Anural development in the ascidian Molgula occulta was examined using tissue-specific markers and interspecific hybridization. Unlike most ascidians, which develop into a swimming tadpole larva (urodele development), M. occulta eggs develop into a tailless slug-like larva (anural development) which metamorphoses into an adult. M. occulta embryos show conventional early cleavage patterns, gastrulation, and neurulation, but then diverge from the urodele developmental mode during larval morphogenesis. M. occulta larvae do not contain a pigmented sensory cell in their brain or form a tail with differentiated notochord and muscle cells. As shown by in situ hybridization with cloned probes and analysis of in vitro translation products, M. occulta embryos do not accumulate high levels of alpha actin or myosin heavy chain mRNA. In contrast, acetylcholinesterase is expressed in muscle lineage cells, indicating that various muscle cell features are differentially suppressed. M. occulta embryos also lack tyrosinase activity, suggesting that suppression of brain pigment cell differentiation occurs at an early step in development. M. occulta eggs fertilized with sperm from Molgula oculata (a closely related urodele species) develop into hybrid larvae exhibiting some of the missing urodele features. Some hybrid embryos develop tyrosinase activity and differentiate a brain pigment cell and a short row of notochord cells, and form a short tail. These urodele features appeared together or separately in different hybrid embryos suggesting that they develop by independent mechanisms. In contrast, alpha actin and myosin heavy chain mRNA accumulation was not enhanced in hybrid embryos. These results suggest that multiple mechanisms control anural development.

Animals↗

PubMed1991

Isolation of cDNAs partially encoding four Xenopus Wnt-1/int-1-related proteins and characterization of their transient expression during embryonic development.

To begin to study the functions of the Wnt-1/int-1 gene family during vertebrate development, we have isolated four Xenopus laevis cDNAs encoding the partial sequence of proteins homologous to Wnt-1/int-1. Xwnt-3, Xwnt-4, Xwnt-5A, and Xwnt-8 demonstrate between 35 and 50% amino acid identity with X. laevis Wnt-1/int-1 and most cysteine residues are conserved. Xwnt-4 and Xwnt-3 transcripts are detected only during the neurula through tadpole stages of development. Expression of Xwnt-8 is observable during gastrulation, declines during neurulation, and is undetectable by the tadpole stage of development. Xwnt-5A transcripts are most prevalent in RNA from oocytes and tadpoles, although low level expression is detected at all stages examined. The temporal changes in expression of these transcripts imply a unique role for each Xwnt during embryogenesis.

Amino Acid Sequence↗

PubMed1991

Modulation of Na,K-ATPase expression during early development of Xenopus laevis.

In amphibian and mammalian systems, regulation of Na+ transport via the Na,K-ATPase plays an important role in distinct developmental processes such as blastocoele formation and neurulation. In this study, we have followed the Na,K-ATPase activity, the biosynthesis, and the cellular accumulation of catalytic alpha-subunits after fertilization of Xenopus laevis eggs up to neurula formation. Our data show that Na,K-ATPase activity increases significantly between stages 4 and 6 and again between stages 13 and 24. The four-fold rise in Na,K-ATPase activity during blastocoele formation is not mediated by an increased cellular pool of alpha-subunits. On the other hand, a five-fold increase of the biosynthesis rate around midblastula precedes a progressive accumulation up to neurula stage mainly of alpha 1-subunits and to a lesser extent of a second alpha-immunoreactive species. In contrast, newly synthesized glycoproteinic beta 1-subunits of Na,K-ATPase cannot be detected up to late neurula. These data indicate that (1) upregulation of Na,K-ATPase activity during blastocoele and neurula formation are mediated by different regulation mechanisms and (2) alpha- and possibly beta-isoforms are expressed in a developmentally regulated fashion during early Xenopus development.

Animals↗

PubMed1988

A third striated muscle actin gene is expressed during early development in the amphibian Xenopus laevis.

During early embryonic development in the frog Xenopus laevis, several muscle-specific actin genes encoding distinct actin protein isoforms are activated in cells of the embryonic muscle. In addition to the cardiac (or alpha 1) and skeletal (or alpha 2) actin genes, a third muscle-specific actin gene is expressed in the same embryonic tissue. We have determined the complete nucleotide sequence of this third gene and examined its expression in embryonic and adult tissues. During embryogenesis, this femoral (alpha 3) actin gene is activated several hours later than its cardiac and skeletal counterparts and its transcripts are first detected after neurulation. The gene encodes a skeletal-type actin protein and is expressed exclusively in skeletal muscle in the adult frog. Two copies of this gene have been isolated from the tetraploid species Xenopus laevis, differing by only a few nucleotides in their protein-coding sequence. The related, diploid species, Xenopus tropicalis, possesses a single copy of the alpha 3 gene and its transcript is similarly conserved in nucleotide sequence. However, the X. tropicalis gene is expressed exclusively in embryonic stages of development. Comparison of the X. laevis and X. tropicalis alpha 3 gene promoters reveals extensive sequence homology, including several copies of a repeated motif that is common to other vertebrate striated-muscle actin gene promoters.

Actins↗

PubMed1983

Calcium, microfilaments and morphogenesis.

Morphogenesis, the generation of tissue form, is important not only in the embryogenesis of a new individual, but also because a change in morphogenesis may be involved in the establishment of differences between individuals during evolution. Morphogenetic movements are effected in part by coordinated changes in the shapes of individual cells and over the past decade the cellular organelles responsible for cell shape have been identified as microfilaments and microtubules. In non-embryonic systems the contraction of microfilaments is controlled by the level of intracellular free calcium, and so calcium is implicated as an intermediate control mechanism in morphogenisis. Through techniques which perturb the calcium balance of cells, or which measure calcium ion concentration directly, evidence is accumulating that calcium is involved in morphogenetic movements such as gastrulation and neurulation, and related phenomena such as wound healing. Thus fundamental questions about the control of morphogenesis in embryogenesis and evolution may now be couched in more precise terms of the control of intracellular calcium ion balance.

Animals↗

PubMed1988

CO2 production of the chick embryo during the first day of post-laying development.

The carbon dioxide production of the chick embryo cultured in vitro has been determined during the first 24 h of post-laying development using a non-invasive conductometric microtechnique. The mean CO2 production of the whole blastoderm (1) increased from 16 nmol/h at laying to 231 nmol/h at early neurulation, (2) became dependent on exogenous glucose and (3) was closely linked to mechanical tension generated in the blastoderm (loosening from vitelline membrane resulted in a decrease of 56%). In our experimental conditions, no significant influence of carbonic anhydrase on the CO2 production has been detected. The value of the respiratory exchange ratio varied from about 3 at pregastrular stages to 1 at neurula stage and CO2 was produced transiently in presence of antimycin A. Such results indicate that the source of CO2 is not exclusively mitochondrial and that the relative proportions of mitochondrial and non-mitochondrial CO2 productions might vary significantly throughout the early development. Our findings confirm that the metabolism of the chick embryo becomes more and more oxidative from laying onwards and suggest that the modifications of metabolism observed during the studied period of development could be associated with functional differentiation.

2,4-Dinitrophenol↗

PubMed1984

The cholinergic system of the primitive streak chick embryo.

The presence of neurotransmitters at stages of embryonic development prior to neurulation has been demonstrated in several systems. Although the functions of these molecules at early stages of embryogenesis have not been ascertained, it is possible that they are involved in aspects of cell migration, regulation of the synthesis of macromolecules, intercellular communication, and in the transmission of positional information during gastrulation. As an initial approach to the resolution of questions concerning the function of transmitters during early development, we have begun a study of the cholinergic system in the primitive streak chick embryo (Hamburger-Hamilton stages 3 + to 5). We have found that the chick embryo: (1) can use exogenously applied choline for the synthesis of acetylcholine; (2) possesses a true acetylcholinesterase, which is predominantly in the form of the 4-6s monomer; and (3) can take up exogenous choline through a sodium-dependent, high-affinity choline transport system. To date we do not have any evidence for the presence of nicotinic or muscarinic receptors at the primitive streak stage.

Acetylcholine↗

PubMed1985

Peanut lectin receptors in the early amphibian embryo: regional markers for the study of embryonic induction.

The regional and temporal specificity of peanut agglutinin binding was determined for early amphibian embryos. With the onset of neurulation, a receptor appears on the epidermis, but remains absent from the neural plate. A second type of receptor, largely masked by sialic acid, appears throughout the extracellular matrix. In the axolotl, the epidermal receptor is epimucin and the matrix receptor is fibronectin plus other components. Both receptors are autonomously expressed, on schedule, by appropriate explants of gastrula tissue. Expression of the epidermal receptor is suppressed after exposure to a neural inducing signal. This shows that the epidermal PNA receptor is a reliable marker of epidermal character and that neural induction affects the program of macromolecular synthesis within hours of the graft.

Ambystoma↗

PubMed1987

Transient activation of oocyte 5S RNA genes in Xenopus embryos by raising the level of the trans-acting factor TFIIIA.

The concentration of the trans-acting factor, TFIIIA, required for the activation of 5S RNA genes in Xenopus can be elevated in developing embryos by injecting a synthetic full-length mRNA into fertilized eggs. 5S RNA genes are activated by the increased factor concentration at the mid-blastula transition through mid-gastrulation. The activated oocyte 5S RNA genes are then inactivated, leaving the TFIIIA-enhanced embryos with the same profile of differential 5S RNA gene activity as control embryos, i.e., synthesizing mainly somatic 5S RNA. Inactivation of the oocyte 5S RNA genes is complete by neurulation and can occur in the absence of DNA replication. We propose that this loss of gene activity is due at least in part to destabilization of transcription complexes that are associated with oocyte 5S RNA genes.

Animals↗

PubMed1995

The identification of two novel ligands of the FGF receptor by a yeast screening method and their activity in Xenopus development.

We have developed a functional screen in yeast to identify ligands for receptor tyrosine kinases. Using this method, we cloned two Xenopus genes that activate the fibroblast growth factor (FGF) receptor. These encode novel secreted proteins, designated FRL1 and FRL2, distantly related to the epidermal growth factor and angiogenin/ribonuclease families, respectively. Both genes activate the FGF receptor in Xenopus oocytes as well as in yeast. Overexpression induces mesoderm and neural-specific genes in Xenopus explants; induction is blocked by a dominant negative inhibitor of the FGF receptor. FRL1 is broadly expressed during gastrulation and neurulation, while FRL2 is expressed principally in the axial mesoderm and brain at later stages. Our results indicate that despite their lack of similarity with FGF, FRL1 and FRL2 are ligands for the FGF receptor that play distinct roles in development.

Amino Acid Sequence↗

PubMed1992

A computerized 2-dimensional vibrating probe for mapping extracellular current patterns.

We describe a computer-assisted 2-dimensional vibrating probe system for mapping endogenous electric current patterns in biological preparations. This system overcomes some of the main limitations of the original 1-dimensional vibrating probe design and adds several new capabilities. Two piezo-electric bender elements mounted perpendicularly are used to vibrate the probe in a circle by applying 2 sine waves (1 to each element) that are 90 degrees out-of-phase with each other. The circular rotation of the probe allows it to detect simultaneously the 2 orthogonal components of a current in the horizontal plane. The voltages measured by the probe are digitized and analyzed by a computer and are used to calculate a current vector. A graphical representation of the current vector is then superimposed on a video image of the experimental preparation. This probe system responds to known currents in the expected manner and exhibits a low inherent noise level. Also included in this paper are some preliminary measurements made with this instrument on neurulating Xenopus embryos and on transected larval sea lamprey (Petromyzon marinus) spinal cords.

Animals↗

PubMed1983

Anomalies of neuroepithelial cell associations in the Splotch mutant embryo.

Neuroepithelial development in Splotch mutant mouse embryos was examined using SEM and TEM. Unlike unaffected littermates, Sp/Sp embryos exhibited extensive neural tube closure defects, disorganization and misalignment of neuroepithelial cells in regions with open defects. Neuroepithelial cell processes often made lateral progress in these areas rather than maintaining their normal contact with the luminal surface. In addition, intercellular space was considerably increased and there were many ectopic cell processes in basal regions of affected areas. The role of altered cell-cell interactions in neurulation is considered.

Animals↗

PubMed1996

Developmental expression of MARCKS and protein kinase C in mice in relation to the exencephaly resulting from MARCKS deficiency.

The roles of protein kinase C and its substrates in development are poorly understood. Recently, we disrupted the mouse gene for a major cellular substrate for protein kinase C, the MARCKS protein (Proc. Natl. Acad. Sci. USA, 92, 944-948, 1995). The resulting phenotype consisted of universal perinatal lethality, agenesis of the corpus callosum and other forebrain commissures, and neuronal ectopia and other cortical and retinal lamination disturbances. These mice also had high frequencies of exencephaly (25% overall, 35% in females). In the present study, we have examined the normal expression of MARCKS and the various isozymes of protein kinase C at the time of cranial neural tube closure, in an attempt to correlate MARCKS expression in time and anatomical location with the exencephaly characteristic of MARCKS deficiency. Failure of neural tube closure occurred at various sites in the cranial neural tube, suggesting a cellular functional defect that was not limited to a specific location. Non-exencephalic MARCKS-deficient embryos appeared to be anatomically normal on embryonic day (E) 8.5-9.5. MARCKS and PKC alpha were expressed at the plasma membrane of the neuroepithelial cells comprising the future neural tube, as well as in the surface ectoderm and underlying mesenchyme. Endogenous protein kinase C species, comprising either or both alpha and delta, were capable of phosphorylating MARCKS in intact E8.5 embryos. Thus, MARCKS is expressed at the plasma membranes of the specific cell types involved in cranial neurulation; its deficiency presumably results in a still-to-be-elucidated functional defect in these cells that leads to exencephaly in a high proportion of cases.

Animals↗

PubMed1994

Cloning of a brain-type aldolase cDNA and changes in its mRNA level during oogenesis and early embryogenesis in Xenopus laevis.

A full length cDNA clone (cXALD3) for Xenopus laevis aldolase mRNA, which exists abundantly in oocytes, was isolated from Xenopus laevis ovary cDNA library, and its nucleotide sequence was determined. The cDNA was 1.8 kb in length and encoded 363 amino acids. From the deduced amino acid sequence and the Northern blot analysis of the RNAs from several adult tissues, this clone was concluded to be a brain-type aldolase gene. The XALD3 mRNA level per egg or embryo was high during early oogenesis, but was markedly reduced during late oogenesis and was maintained at low level during early embryogenesis until it started to increase at the late neurula stage. The mRNA was also detected in testis. The characteristic change in the temporal pattern of expression and the distribution of XALD3 mRNA among different tissues suggest a possibility that brain type aldolase may play some important roles in gametogenesis and in neurulation.

Amino Acid Sequence↗

PubMed1992

Retinoic acid-induced neural tube defects with multiple canals in the chick: immunohistochemistry with monoclonal antibodies.

When retinoic acid was injected into chicken yolks before incubation, various types of neural tube defect (NTD) were induced in 38-46% of the embryos after 48-96 h of incubation. The cranial NTD consisted of a delay in closing of the neural plate in 48-h embryos and some remained as disorganized, hyperplastic masses in older embryos. In spinal NTD of 48-h embryos the posterior neuropore remained widely open. In older embryos with a closed posterior neuropore, the neural tube appeared dissociated or disorganized locally at the trunk level. The tissue consisted of a dorsally-situated, neural-plate-like structure and a ventrally-located cell mass containing multiple canals. Although the location was different, this arrangement was similar to the overlap zone which appears between primary and secondary neurulation in normal development. Immunohistochemistry was performed using monoclonal antibodies which selectively stained various components of chick tissue. Considering the similarity in neural tube formation between chick and human, this experimental NTD may provide clues to understanding the etiology of human myelomeningocele.

Animals↗

PubMed1992

Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space.

Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we intend to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.

Animals↗

PubMed1982

The action of low-intensity pulsed ultrasound on amphibian embryonic tissues.

Amphibian embryos at different stages of development, and ectomesodermal explants (small pieces of embryonic tissue extirpated from lateral parts of the embryos just after neurulation), have been irradiated with continuous and pulsed ultrasound at a frequency of 0.88 MHz, spatial and temporal average intensities being within the range 0.025-0.1 W cm2. Ultrasound pulse repetition frequencies were varied within 10-1000 Hz, duty factor being constant and equal to 0.5. Destruction of embryonic tissue as an immediate effect, and interruption or distortion of development as delayed (24 hr) effects, were registered. Significant damaging effects of non-cavitational and non-thermal origin and dependent on pulse repetition frequencies, were observed. Highly frequency-dependent resonance-type effects were obtained for irradiated explants. The maximum effect on Rana temporaria explants was obtained at pulse repetition frequencies of 10-20 Hz and on Xenopus laevis explants at 110-130 Hz.

Animals↗