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[
1984]
Developmental fates of blastomeres in early C. elegans embryos appear to be governed by internally segregating, cell-autonomous determinants. To ascertain whether previously described gut-lineage dterminants are nuclear or cytoplasmic, laser microsurgery was used to show that exposing the nucleus of a non-gut-precursor cell to gut-precursor cytoplasm can cause the progeny of the resulting hybrid cell to express gut-specific differentiation markers, supporting the view that the determinants are cytoplasmic. In attempts to obtain molecular probes for such determinants, a library of monoclonal antibodies to early embryonic antigens was generated and screened by immunofluorescence microscopy for antibodies reacting with lineage-specific components. Three of the antibodies react with cytoplasmic granules (P granules) that segregate specifically with the germ line in early cleavages and are found uniquely in germ-line cells throughout the life cycle. Experiments on unfertilized eggs, on mutant embryos with defects in early cleavage, and on normal embryos treated with various cytoskeletal inhibitors indicate that P-granule segregation depends upon fertilization and requires the function of actin microfilaments, but is independent of spindle and microtubule functions. Work on the biochemical nature and function of the P granules is in progress.
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[
1987]
We describe the use of a nonspecific carboxylesterase as a biochemical marker for intestinal differentiation in the nematode C. elegans. In particular, we describe how esterase expression responds to inhibition of embryonic DNA synthesis by aphidicolin. Esterase expression requires a short period of DNA synthesis immediattely after the gut lineage is clonally established. However, the subsequent 2-3 rounds of DNA synthesis, which normally occur before esterase gene transcription, can be inhibited without effect. Thus esterase expression depends neither on reaching the normal DNA:cytoplasmic ration nor on counting the normal number of replication rounds.
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[
1987]
We describe an experimental system in which to study gene-specific segregation mechanisms during early development of C. elegans. A non-specific esterase, of unknown physiological function, has convenient properties as a biochemical marker of differentiation: expression is localized to the gut lineage, is due to transcription during zygotic development and is lineage autonomous. The timing of esterase expression does not depend either on the normal number of rounds of cytokinesis or on the normal number of rounds of DNA replication; thus some other clock mechanism must be invoked. We descrbe experiments suggesting that DNA strands donated by the sperm do not co-segregate during development of the next generation.
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[
WormBook,
2007]
The C. elegans foregut (pharynx) has emerged as a powerful system to study organ formation during embryogenesis. Here I review recent advances regarding cell-fate specification and epithelial morphogenesis during pharynx development. Maternally-supplied gene products function prior to gastrulation to establish pluripotent blastomeres. As gastrulation gets under way, pharyngeal precursors become committed to pharyngeal fate in a process that requires PHA-4 /FoxA and the Tbox transcription factors TBX-2 , TBX-35 , TBX-37 and TBX-38 . Subsequent waves of gene expression depend on the affinity of PHA-4 for its target promoters, coupled with combinatorial strategies such as feed-forward and positive-feedback loops. During later embryogenesis, pharyngeal precursors undergo reorganization and a mesenchymal-to-epithelial transition to form the linear gut tube. Surprisingly, epithelium formation does not depend on cadherins, catenins or integrins. Rather, the kinesin ZEN-4 /MKLP1 and CYK-4 /RhoGAP are critical to establish the apical domain during epithelial polarization. Finally, I discuss similarities and differences between the nematode pharynx and the vertebrate heart.
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[
1979]
In many invertebrates, cell lineages are apparently invariant from individual to individual. A given precursor cell follows a specific pattern of cell divisions, and its descendants follow fates that correspond to their respective positions in the lineage tree. Such a reproducible sequence of events provides an excellent system for studying how cells come to pursue particular fates during development. We have been interested to know if a cell's fate is specified by factors intrinsic to the cell, or if it is influenced by interactions between the cell and its environment. C. elegans is a particularly suitable organism for lineage studies because it is transparent throughout its life cycle, and because it consists of relatively few cells. Furthermore, C. elegans is a favorable organism for genetics, so the control of cell lineages can be studied by characterizing mutations that are defective in known lineages. The cell lineages of C. elegans have been described in the embryo to the 182 cell stage and after hatching. Approximately 50 cells resume divisions post-embyronically. In the somatic tissues, the number of cells (or nuclei) is increased from about 550 to about 950 in hermaphrodites and to about 1025 in males. These post-embryonic lineages are essentially invariant from worm to worm. As the worm enlarges and matures sexually, cells (or nuclei) are added to previously existing tissues (hypodermis, muscle, gut, and nervous system), and structures necessary for reproduction are elaborated. The latter include a gonad in both sexes, a vulva in hermaphrodites, and a tail specialized for copulation in males. This paper summarizes the results of laser ablation experiments performed on cells in the post-embryonic lineages of C. elegans. In particular, we focus on those experiments that demonstrate a regulative capacity in the cells of this predominantly invariant system. The post-embyronic lineages have the practical advantage for these studies that they can be traced by direct observation of the cells as they divide and assume their final fate. The regulative response, therefore, can be described at a level of cellular detail that has not been possible in other deletion studies. Our aim in performing these experiments is to infer how cells are controlled during normal development from their behavior in