Cytoplasmic calcium gradients and calmodulin in the early development of the fucoid alga Pelvetia compressa

1998 ◽  
Vol 111 (21) ◽  
pp. 3197-3207 ◽  
Author(s):  
R. Pu ◽  
K.R. Robinson
Keyword(s):  
Rhodamine B ◽  
Normal Development ◽  
Excitation Wavelength ◽  
Cytoplasmic Calcium ◽  
Calmodulin Antagonist ◽  
Early Embryos ◽  
Pelvetia Compressa ◽  
Photosensitive Period ◽  
Early Developmental ◽  
Rhizoid Formation

The predicted existence of cytoplasmic Ca2+ gradients during the photopolarization of the zygotes of the brown algae, Pelvetia and Fucus, has proved to be difficult to establish, and the downstream targets of the putative gradients are not known. We have used quantitative microinjection of the long excitation wavelength Ca2+ indicator, Calcium Crimson, and of antibodies against calmodulin to investigate these matters in the zygotes and early embryos of Pelvetia. We found that there is a window of cytoplasmic Calcium Crimson concentration that gives an adequate signal above autofluorescence yet allows normal development of the zygotes. As Calcium Crimson is not a ratiometric indicator, we injected other zygotes with a Ca2+-insensitive dye, rhodamine B, and imaged the cells at the same time that Calcium Crimson-injected cells were imaged. Ratios were calculated by dividing the averaged pixel values of Calcium Crimson images by the averaged pixel values of corresponding rhodamine B images. By this method, we observed the formation of a cytoplasmic Ca2+ gradient within one hour of the exposure of the cells to unilateral blue light during the photosensitive period. The region of high Ca2+ was localized to and predictive of the site of future rhizoid formation. We validated this somewhat indirect method by applying it to the growing rhizoid, where the existence of a tip-localized Ca2+ gradient is well established. The method clearly revealed the known gradient. The injection of ungerminated zygotes with antibodies made against Dictyostelium calmodulin inhibited germination, and this inhibition was abolished if the calmodulin antibodies were coinjected with an excess of purified maize calmodulin. Likewise, the growth of the rhizoids was inhibited by calmodulin antibody injections. The fungus-derived calmodulin antagonist, ophiobolin A, which has previously been shown to be a potent inhibitor of germination, also inhibited rhizoidal growth. Our results provide evidence that a cytoplasmic Ca2+ gradient is present during photopolarization and that calmodulin acts as a mediator of Ca2+ gradients throughout the early developmental processes of germination and rhizoidal growth in Pelvetia compressa.

scholarly journals Dnmt1 binds and represses genomic retroelements via DNA methylation in mouse early embryos

2020 ◽  
Vol 48 (15) ◽  
pp. 8431-8444 ◽  
Author(s):  
Byungkuk Min ◽  
Jung Sun Park ◽  
Young Sun Jeong ◽  
Kyuheum Jeon ◽  
Yong-Kook Kang
Keyword(s):  
Dna Methylation ◽  
Cpg Islands ◽  
Endogenous Retrovirus ◽  
Dna Demethylation ◽  
Mouse Development ◽  
Genome Wide ◽  
Early Embryos ◽  
Dna Adenine Methylase ◽  
Early Mouse ◽  
Early Developmental

Abstract Genome-wide passive DNA demethylation in cleavage-stage mouse embryos is related to the cytoplasmic localization of the maintenance methyltransferase DNMT1. However, recent studies provided evidences of the nuclear localization of DNMT1 and its contribution to the maintenance of methylation levels of imprinted regions and other genomic loci in early embryos. Using the DNA adenine methylase identification method, we identified Dnmt1-binding regions in four- and eight-cell embryos. The unbiased distribution of Dnmt1 peaks in the genic regions (promoters and CpG islands) as well as the absence of a correlation between the Dnmt1 peaks and the expression levels of the peak-associated genes refutes the active participation of Dnmt1 in the transcriptional regulation of genes in the early developmental period. Instead, Dnmt1 was found to associate with genomic retroelements in a greatly biased fashion, particularly with the LINE1 (long interspersed nuclear elements) and ERVK (endogenous retrovirus type K) sequences. Transcriptomic analysis revealed that the transcripts of the Dnmt1-enriched retroelements were overrepresented in Dnmt1 knockdown embryos. Finally, methyl-CpG-binding domain sequencing proved that the Dnmt1-enriched retroelements, which were densely methylated in wild-type embryos, became demethylated in the Dnmt1-depleted embryos. Our results indicate that Dnmt1 is involved in the repression of retroelements through DNA methylation in early mouse development.

scholarly journals Microarray analysis of gene expression during early development: a cautionary overview

Reproduction ◽  
2010 ◽  
Vol 140 (6) ◽  
pp. 787-801 ◽  
Author(s):  
Claude Robert
Keyword(s):  
Gene Expression ◽  
Early Development ◽  
Microarray Analysis ◽  
Developmental Stages ◽  
Sample Handling ◽  
Early Embryos ◽  
High Throughput Method ◽  
Full Benefit ◽  
Key Steps ◽  
Early Developmental

The rise of the ‘omics’ technologies started nearly a decade ago and, among them, transcriptomics has been used successfully to contrast gene expression in mammalian oocytes and early embryos. The scarcity of biological material that early developmental stages provide is the prime reason why the field of transcriptomics is becoming more and more popular with reproductive biologists. The potential to amplify scarce mRNA samples and generate the necessary amounts of starting material enables the relative measurement of RNA abundance of thousands of candidates simultaneously. So far, microarrays have been the most commonly used high-throughput method in this field. Microarray platforms can be found in a wide variety of formats, from cDNA collections to long or short oligo probe sets. These platforms generate large amounts of data that require the integration of comparative RNA abundance values in the physiological context of early development for their full benefit to be appreciated. Unfortunately, significant discrepancies between datasets suggest that direct comparison between studies is difficult and often not possible. We have investigated the sample-handling steps leading to the generation of microarray data produced from prehatching embryo samples and have identified key steps that significantly impact the downstream results. This review provides a discussion on the best methods for the preparation of samples from early embryos for microarray analysis and focuses on the challenges that impede dataset comparisons from different platforms and the reasons why methodological benchmarking performed using somatic cells may not apply to the atypical nature of prehatching development.

scholarly journals Variability of an early developmental cell population underlies stochastic laterality defects

2020 ◽  
Author(s):  
Roberto Moreno-Ayala ◽  
Pedro Olivares-Chauvet ◽  
Ronny Schäfer ◽  
Jan Philipp Junker
Keyword(s):  
Elevated Temperature ◽  
Maternal Effects ◽  
Genetic Background ◽  
Normal Development ◽  
Cell Number ◽  
Vertebrate Development ◽  
Stochastic Variation ◽  
Number Variation ◽  
Early Developmental ◽  
Laterality Defects

Embryonic development seemingly proceeds with almost perfect precision. However, it is largely unknown how much underlying microscopic variability is compatible with normal development. Here, we quantified embryo-to-embryo variability in vertebrate development, by studying cell number variation in the zebrafish endoderm. We noticed that the size of a sub- population of the endoderm, the dorsal forerunner cells (which later form the left-right organizer), exhibits significantly more embryo-to-embryo variation than the rest of the endoderm. We found that, when incubated at elevated temperature, the frequency of left-right laterality defects is increased drastically in embryos with a low number of dorsal forerunner cells. Furthermore, we observed that these fluctuations have a large stochastic component among fish of the same genetic background. Hence, a stochastic variation in early development leads to a remarkably strong macroscopic phenotype. These fluctuations appear to be associated with maternal effects in the specification of the dorsal forerunner cells.

scholarly journals Physical constraints on early blastomere packings

2021 ◽  
Vol 17 (1) ◽  
pp. e1007994
Author(s):  
James Giammona ◽  
Otger Campàs
Keyword(s):  
Cell Division ◽  
Developmental Stages ◽  
Physical Parameters ◽  
Cell Stage ◽  
Physical Constraints ◽  
Equilibrium Dynamics ◽  
Deformable Particles ◽  
Early Embryos ◽  
Early Developmental Stages ◽  
Early Developmental

At very early embryonic stages, when embryos are composed of just a few cells, establishing the correct packing arrangements (contacts) between cells is essential for the proper development of the organism. As early as the 4-cell stage, the observed cellular packings in different species are distinct and, in many cases, differ from the equilibrium packings expected for simple adherent and deformable particles. It is unclear what are the specific roles that different physical parameters, such as the forces between blastomeres, their division times, orientation of cell division and embryonic confinement, play in the control of these packing configurations. Here we simulate the non-equilibrium dynamics of cells in early embryos and systematically study how these different parameters affect embryonic packings at the 4-cell stage. In the absence of embryo confinement, we find that cellular packings are not robust, with multiple packing configurations simultaneously possible and very sensitive to parameter changes. Our results indicate that the geometry of the embryo confinement determines the packing configurations at the 4-cell stage, removing degeneracy in the possible packing configurations and overriding division rules in most cases. Overall, these results indicate that physical confinement of the embryo is essential to robustly specify proper cellular arrangements at very early developmental stages.

scholarly journals Chromosome damage and early developmental arrest caused by the Rex element of Drosophila melanogaster.

Genetics ◽  
1994 ◽  
Vol 138 (2) ◽  
pp. 401-411 ◽  
Author(s):  
L G Robbins ◽  
S Pimpinelli
Keyword(s):  
Drosophila Melanogaster ◽  
Ribosomal Dna ◽  
Maternal Effect ◽  
Cell Biology ◽  
Chromosome Breakage ◽  
Chromosome Damage ◽  
Specific Chromosome ◽  
Developmental Arrest ◽  
Early Embryos ◽  
Early Developmental

Abstract Rex (Ribosomal exchange) is a genetically identified repeated element within the ribosomal DNA (rDNA) of Drosophila melanogaster. Rex has a semidominant maternal effect that promotes exchange between and within rDNA arrays in the first few embryonic mitoses. Several of Rex's genetic properties suggest that its primary effect is rDNA-specific chromosome breakage that is resolved by recombination. We report here that rDNA crossovers are only a small, surviving minority of Rex-induced events. Cytology of embryos produced by Rex-homozygous females reveals obvious chromosome damage in at least a quarter of the embryos within the first three mitotic divisions. More than half of the embryos produced by Rex females die, and the developmental arrest is among the earliest reported for any maternal-effect lethal. The striking lethal phenotype suggests that embryos with early chromosome damage could be particularly fruitful subjects for analysis of the cell biology of early embryos.

scholarly journals Carl-Philipp Heisenberg: Early embryos make a big move

2013 ◽  
Vol 200 (3) ◽  
pp. 238-239
Author(s):  
Caitlin Sedwick
Keyword(s):  
Physical Properties ◽  
Early Embryos ◽  
Early Developmental

Heisenberg is studying how cells’ physical properties drive early developmental events.

Normal development and embryonic gene activity of the ascidian Herdmania momus

1996 ◽  
Vol 47 (3) ◽  
pp. 543 ◽  
Author(s):  
BM Degnan ◽  
PR Rohde ◽  
MF Lavin
Keyword(s):  
Normal Development ◽  
Homeobox Genes ◽  
Cell Movement ◽  
Model Systems ◽  
Temperature Dependent ◽  
Cell Stage ◽  
Early Embryos ◽  
Tailbud Stage ◽  
Solitary Ascidian ◽  
Early Gastrula

Embryonic and post-larval development of the tropical solitary ascidian Herdmania momus is shown to be similar to that of extensively studied ascidian model systems. H. momus development is rapid and temperature-dependent, with hatching occurring 8.5 h after fertilization at 28�C. An increase in total embryonic gene transcription is detected at the 110-cell stage or the onset of gastrulation. Treatment of early embryos with actinomycin D inhibits transcription and curtails morphogenetic cell movement in the early gastrula without immediately inhibiting cell division. The prevalence of homeobox-containing transcripts increases around the 110-cell stage and later in development. Isolated H. momus homeobox genes, expressed at the tailbud stage, have greatest sequence identity to members of Hox, otd/Otx, eve/Evx and cad/Cdx homeobox classes. Evidence from H. momus and other ascidians suggests that urochordates possess most of the homeobox genes of the chordate HOX cluster.

scholarly journals Cyclin D and cdk4 Are Required for Normal Development beyond the Blastula Stage in Sea Urchin Embryos

2002 ◽  
Vol 22 (13) ◽  
pp. 4863-4875 ◽  
Author(s):  
Jennifer C. Moore ◽  
Jan L. Sumerel ◽  
Bradley J. Schnackenberg ◽  
Jason A. Nichols ◽  
Athula Wikramanayake ◽  
...  
Keyword(s):  
Sea Urchin ◽  
Normal Development ◽  
Sea Urchins ◽  
Ectopic Expression ◽  
Cyclin D ◽  
Blastula Stage ◽  
Cell Stage ◽  
Cell Cycles ◽  
Early Embryos ◽  
Cdk4 Expression

ABSTRACT cdk4 mRNA and protein are constitutively expressed in sea urchin eggs and throughout embryonic development. In contrast, cyclin D mRNA is barely detectable in eggs and early embryos, when the cell cycles consist of alternating S and M phases. Cyclin D mRNA increases dramatically in embryos at the early blastula stage and remains at a constant level throughout embryogenesis. An increase in cdk4 kinase activity occurs concomitantly with the increase in cyclin D mRNA. Ectopic expression of cyclin D mRNA in eggs arrests development before the 16-cell stage and causes eventual embryonic death, suggesting that activation of cyclin D/cdk4 in cleavage cell cycles is lethal to the embryo. In contrast, blocking cyclin D or cdk4 expression with morpholino antisense oligonucleotides results in normal development of early gastrula-stage embryos but abnormal, asymmetric larvae. These results suggest that in sea urchins, cyclin D and cdk4 are required for normal development and perhaps the patterning of the developing embryo, but may not be directly involved in regulating entry into the cell cycle.

scholarly journals Effect of heritable symbionts on maternally-derived embryo transcripts

2019 ◽  
Author(s):  
Mariana Mateos ◽  
Nadisha O. Silva ◽  
Paulino Ramirez ◽  
Victor M. Higareda-Alvear ◽  
Rodolfo Aramayo ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Cytoplasmic Incompatibility ◽  
Developmental Stages ◽  
Early Embryo ◽  
Male Embryo ◽  
Ribosome Inactivating Proteins ◽  
Male Killing ◽  
Early Embryos ◽  
Early Developmental ◽  
Mrna Transcriptome

AbstractMaternally-transmitted endosymbiotic bacteria are ubiquitous in insects. Among other influential phenotypes, many heritable symbionts of arthropods are notorious for manipulating host reproduction through one of four reproductive syndromes, which are generally exerted during early developmental stages of the host: male feminization; parthenogenesis induction; male killing; and cytoplasmic incompatibility (CI). Major advances have been achieved in understanding mechanisms and identifying symbiont factors involved in reproductive manipulation, particularly male killing and cytoplasmic incompatibility. Nonetheless, whether cytoplasmically-transmitted bacteria influence the maternally-loaded components of the egg or early embryo has not been examined. In the present study, we investigated whether heritable endosymbionts that cause different reproductive phenotypes in Drosophila melanogaster influence the mRNA transcriptome of early embryos. We used mRNA-seq to evaluate differential expression in Drosophila embryos lacking endosymbionts (control) to those harbouring the male-killing Spiroplasma poulsonii strain MSRO-Br, the CI-inducing Wolbachia strain wMel, or Spiroplasma poulsonii strain Hyd1; a strain that lacks a reproductive phenotype and is naturally associated with Drosophila hydei. We found no consistent evidence of influence of symbiont on mRNA composition of early embryos, suggesting that the reproductive manipulation mechanism does not involve alteration of maternally-loaded transcripts. In addition, we capitalized on several available mRNA-seq datasets derived from Spiroplasma-infected Drosophila melanogaster embryos, to search for signals of depurination of rRNA, consistent with the activity of Ribosome Inactivating Proteins (RIPs) encoded by Spiroplasma poulsonii. We found small but statistically significant signals of depurination of Drosophila rRNA in the Spiroplasma treatments (both strains), but not in the symbiont-free control or Wolbachia treatment, consistent with the action of RIPs. The depurination signal was slightly stronger in the treatment with the male-killing strain. This result supports a recent report that RIP-induced damage contributes to male embryo death.

Analysis of embryonic induction by using cell lineage markers

1984 ◽  
Vol 307 (1132) ◽  
pp. 331-336 ◽  
Keyword(s):  
Normal Development ◽  
Cell Lineage ◽  
Neural Induction ◽  
Mesoderm Induction ◽  
Target Tissue ◽  
Single Cell Level ◽  
Embryonic Induction ◽  
Cell Level ◽  
Early Embryos ◽  
Lineage Markers

Three distinct inductive interactions have been demonstrated in early embryos of Xenopus laevis :mesoderm induction, dorsalization and neural induction. The experiments were done with grafts from embryos uniformly labelled with passive cell lineage markers, either FITC-lysine-dextran (FLDx) or horseradish peroxidase (HRP), which allow the provenance of regions to be determined down to the single cell level. In each case the fate of the target tissue in the presence of the appropriate inductor was quite different from the fate in normal development.

Sign in / Sign up

Export Citation Format

Share Document