- Caenorhabditis elegans as a Genomic Model for Toxicology
Effective toxicological testing of the vast number of new and existing chemicals currently in use will require efficient and cost effective methods. We evaluated the utility of a simple, low cost toxicity testing system employing the nematode Caenorhabditis elegans to identify toxicologically relevant changes in gene expression. Dichlorvos and fenamiphos, which are organophosphorous pesticides that inhibit acetylcholinesterase were chosen as model toxicants to test the usefulness of the C. elegans toxicity testing system, and mefloquine, which appears to perturb neuronal Ca++ homeostasis, provided an out-group for analysis. Keywords: gene expression array-based (RNA / in situ oligonucleotide)
- Analysis of SDC-2, DPY-26, MIX-1, DPY-27 and RNA Polymerase II ChIP-chip and RNA abundance in C. elegans
In C. elegans, a condensin-like protein complex associates specifically with both X chromosomes of XX animals to execute dosage compensation. Dosage Compensation Complex (DCC) reduces the level of transcripts arising from each of the two X chromosomes. Recruitment to X is specified in part by discrete DNA sequence motifs, but following recruitment, the DCC is targeted to the promoters of individual active genes by an unknown mechanism. Here, we investigated three outstanding questions regarding the molecular mechanism of DCC recruitment and spreading along X. By examining the genome-wide binding patterns of several DCC subunits in different stages of C. elegans development, and in strains harboring X:Autosome chromosomal fusions, we provide evidence that: (1) DCC binding is dynamically specified according to gene activity during development (2) The condensin-like subunits of the DCC spread from recruitment sites to active promoters more readily than the non-conserved SDC subunits, which are involved in initial X-targeting and (3) the mechanism of DCC spreading is independent of X-chromosome DNA sequence, and will proceed onto any active promoter near a recruitment site. Our results contribute to understanding how chromatin complexes can be targeted to achieve domain-scale transcriptional regulation during development. For data usage terms and conditions, please refer to http:
- Heme homeostasis is regulated by the conserved and concerted functions of HRG-1 proteins
Hemes are essential but potentially cytotoxic cofactors that participate in critical and diverse biological processes. Although the pathway and intermediates for heme biosynthesis have been well defined, the intracellular networks which mediate heme trafficking remain unknown. Caenorhabditis elegans and related helminths are natural heme auxotrophs requiring environmental heme for growth and development. We exploited this auxotrophy to identify HRG-1 and HRG-4 in C. elegans and show that they are essential for heme homeostasis and normal vertebrate development. We demonstrate that heme deficiency upregulates expression of
hrg-4 and its evolutionarily conserved paralog
hrg-1. Depletion of either HRG-1 or HRG-4 in worms results in disruption of organismal heme sensing and abnormal response to heme analogs. HRG-1 and HRG-4 are novel transmembrane proteins that bind heme and have evolutionarily conserved functions. Transient knockdown of
hrg-1 in zebrafish leads to hydrocephalus, yolk tube malformations, and, most strikingly, profound defects in erythropoiesis - phenotypes that are fully rescued by worm HRG-1. These findings reveal unanticipated and conserved pathways for cellular heme trafficking in animals that defines the paradigm for eukaryotic heme transport. Uncovering the mechanisms of heme transport in C. elegans will provide novel insights into human disorders of heme metabolism and generate unique anthelmintics to combat worm infestations. Keywords: dose-response
- Developmental milestones punctuate gene expression in the Caenorhabditis embryo
The complex process by which a metazoan develops from a single cell to a multi-cellular differentiated organism is typically organized by biologists into stages. For example, in the chordate embryo, the gastrula, the neurula and the tail-bud constitute characteristic processes. A debate runs through the history of embryology questioning whether such stages hint at the modularity of embryonic development1-3, or rather, that stages exist in the mind of the biologist, and not in the larva4 or embryo. It thus remains unclear which position accurately reflects the nature of development on a molecular, gene regulatory level. Here we demonstrate that development of five Caenorhabditis species proceeds through multiple distinct stages in which the transcriptome is resistant to differences in species-specific developmental timings. By comparing the complete protein-coding transcriptomes of individually staged embryos across ten morphological markers, we found that time-invariant stages occur throughout development, including a stage we identify as the nematode phylotypic stage. Between such stages, embryos follow transitory states characterized by an acceleration of transcriptional activity. Therefore, on a molecular level, development cannot be viewed as a single process continuously proceeding through time, but rather as a succession of discontinuous stages, or milestones. Comparing the nematode developmental transcriptome with that of the chordate we report on the macro evolution of a milestone by duplication and diversification. This modular view of development by milestones will allow for a more complete understanding of how the functional organization of the embryo has influenced the evolution of animal morphology and diversity.
- Developmental milestones punctuate gene expression in the Caenorhabditis embryo
The complex process by which a metazoan develops from a single cell to a multi-cellular differentiated organism is typically organized by biologists into stages. For example, in the chordate embryo, the gastrula, the neurula and the tail-bud constitute characteristic processes. A debate runs through the history of embryology questioning whether such stages hint at the modularity of embryonic development1-3, or rather, that stages exist in the mind of the biologist, and not in the larva4 or embryo. It thus remains unclear which position accurately reflects the nature of development on a molecular, gene regulatory level. Here we demonstrate that development of five Caenorhabditis species proceeds through multiple distinct stages in which the transcriptome is resistant to differences in species-specific developmental timings. By comparing the complete protein-coding transcriptomes of individually staged embryos across ten morphological markers, we found that time-invariant stages occur throughout development, including a stage we identify as the nematode phylotypic stage. Between such stages, embryos follow transitory states characterized by an acceleration of transcriptional activity. Therefore, on a molecular level, development cannot be viewed as a single process continuously proceeding through time, but rather as a succession of discontinuous stages, or milestones. Comparing the nematode developmental transcriptome with that of the chordate we report on the macro evolution of a milestone by duplication and diversification. This modular view of development by milestones will allow for a more complete understanding of how the functional organization of the embryo has influenced the evolution of animal morphology and diversity.
- Developmental milestones punctuate gene expression in the Caenorhabditis embryo
The complex process by which a metazoan develops from a single cell to a multi-cellular differentiated organism is typically organized by biologists into stages. For example, in the chordate embryo, the gastrula, the neurula and the tail-bud constitute characteristic processes. A debate runs through the history of embryology questioning whether such stages hint at the modularity of embryonic development1-3, or rather, that stages exist in the mind of the biologist, and not in the larva4 or embryo. It thus remains unclear which position accurately reflects the nature of development on a molecular, gene regulatory level. Here we demonstrate that development of five Caenorhabditis species proceeds through multiple distinct stages in which the transcriptome is resistant to differences in species-specific developmental timings. By comparing the complete protein-coding transcriptomes of individually staged embryos across ten morphological markers, we found that time-invariant stages occur throughout development, including a stage we identify as the nematode phylotypic stage. Between such stages, embryos follow transitory states characterized by an acceleration of transcriptional activity. Therefore, on a molecular level, development cannot be viewed as a single process continuously proceeding through time, but rather as a succession of discontinuous stages, or milestones. Comparing the nematode developmental transcriptome with that of the chordate we report on the macro evolution of a milestone by duplication and diversification. This modular view of development by milestones will allow for a more complete understanding of how the functional organization of the embryo has influenced the evolution of animal morphology and diversity.
- Developmental milestones punctuate gene expression in the Caenorhabditis embryo
The complex process by which a metazoan develops from a single cell to a multi-cellular differentiated organism is typically organized by biologists into stages. For example, in the chordate embryo, the gastrula, the neurula and the tail-bud constitute characteristic processes. A debate runs through the history of embryology questioning whether such stages hint at the modularity of embryonic development1-3, or rather, that stages exist in the mind of the biologist, and not in the larva4 or embryo. It thus remains unclear which position accurately reflects the nature of development on a molecular, gene regulatory level. Here we demonstrate that development of five Caenorhabditis species proceeds through multiple distinct stages in which the transcriptome is resistant to differences in species-specific developmental timings. By comparing the complete protein-coding transcriptomes of individually staged embryos across ten morphological markers, we found that time-invariant stages occur throughout development, including a stage we identify as the nematode phylotypic stage. Between such stages, embryos follow transitory states characterized by an acceleration of transcriptional activity. Therefore, on a molecular level, development cannot be viewed as a single process continuously proceeding through time, but rather as a succession of discontinuous stages, or milestones. Comparing the nematode developmental transcriptome with that of the chordate we report on the macro evolution of a milestone by duplication and diversification. This modular view of development by milestones will allow for a more complete understanding of how the functional organization of the embryo has influenced the evolution of animal morphology and diversity.
- Developmental milestones punctuate gene expression in the Caenorhabditis embryo
The complex process by which a metazoan develops from a single cell to a multi-cellular differentiated organism is typically organized by biologists into stages. For example, in the chordate embryo, the gastrula, the neurula and the tail-bud constitute characteristic processes. A debate runs through the history of embryology questioning whether such stages hint at the modularity of embryonic development1-3, or rather, that stages exist in the mind of the biologist, and not in the larva4 or embryo. It thus remains unclear which position accurately reflects the nature of development on a molecular, gene regulatory level. Here we demonstrate that development of five Caenorhabditis species proceeds through multiple distinct stages in which the transcriptome is resistant to differences in species-specific developmental timings. By comparing the complete protein-coding transcriptomes of individually staged embryos across ten morphological markers, we found that time-invariant stages occur throughout development, including a stage we identify as the nematode phylotypic stage. Between such stages, embryos follow transitory states characterized by an acceleration of transcriptional activity. Therefore, on a molecular level, development cannot be viewed as a single process continuously proceeding through time, but rather as a succession of discontinuous stages, or milestones. Comparing the nematode developmental transcriptome with that of the chordate we report on the macro evolution of a milestone by duplication and diversification. This modular view of development by milestones will allow for a more complete understanding of how the functional organization of the embryo has influenced the evolution of animal morphology and diversity.
- SGF3165_FLAG_MES4FLAG_EEMB
The focus of our analysis will be elements that specify nucleosome positioning and occupancy, control domains of gene expression, induce repression of the X chromosome, guide mitotic segregation and genome duplication, govern homolog pairing and recombination during meiosis, and organize chromosome positioning within the nucleus. 126 strategically selected targets include key histone modifications, histone variants, RNA polymerase II isoforms, dosage-compensation proteins, centromere components, homolog-pairing facilitators, recombination markers, and nuclear-envelope constituents.We will integrate information generated with existing knowledge on the biology of the targets, perform ChIP-chip analysis on mutant and RNAi extracts lacking selected target proteins, use extrachromosomal arrays to assess the ability of candidate identified sequence motifs to recruit targets in vivo, identify tissue-specific patterns of selected targets, and create integrated, quantitative models of transcription and whole-chromosome function