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Recombinant DNA technology has made it possible to clone receptors from many organisms by cross-hybridization or by the polymerase chain reaction. It may be difficult, though, to establish the functional importance of any clone obtained. We describe the cloning of nematode acetylcholine receptor genes by selection for resistance to levamisole, a scheme providing assurance that the clones obtained are functionally related...
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[
1979]
We have isolated temperature sensitive maternal effect mutants in the free-living nematode Caenorhabditis elegans. We use C. elegans for several basic reasons. It is easy to culture in the laboratory and it has a rapid life cycle. The genetics of C. elegans have been elucidated by Brenner and more recently have been refined by the lethal analysis of Herman et. al. Both embryonic and postembryonic development can be observed directly and conveniently on the living worm with Nomarski differential interference optics because egg shell and worm cuticle are transparent. The precise embryonic cell lineages of C. elegans are known from fertilization to the 200 blastomere stage. All of the postembryonic somatic cell lineages are precisely known. It ...
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[
1981]
A neuron can be characterized by its morphology, transmitter (s?), receptor(s) and the nature of its synaptic contacts (chemical or electrical; excitatory or inhibitory; number and distribution of synapses; identity of the cells to which it is presynaptic or postsynaptic). It is clear that according to such criteria nervous sytems consist of neurons of many distinct types. The origin of neuronal diversity is unknown. Both how such diversity is generated during development and how the relevant developmental programme is encoded in the genome remain to
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[
1982]
Caenorhabditis elegans is a free-living, nonparasitic nematode. It is a self-fertilizing hermaphrodite. Males arise spontaneously by nondisjunction of X-chromosomes. Of all eukaryotic organisms C. elegans has probably been most extensively studied at the cellular level. Within 12 hours the fertilized egg develops into a young larva with 558 nuclei (560 in the male). During postembryonic development the animal proceeds through four larval stages increasing its number of nuclei to 959 (1,031 in the male) plus some 2,000 germ cells (about 1,000 in the male). The cell lineages from fertilization to adulthood have been completely analyzed in living embryos and animals. This and its well-established genetics (more than 300 genes have been mapped on the six linkage groups) make it a suitable model organism to study problems of gene action and development. Various techniques have been used to interfere with normal development (including laser-induced cell ablations) and to analyze development on the subcellular level (including recombinant DNA technology). The characteristic features of rigidly determined development, the low cell number, and the knowledge of cellular events should make it possible to identify molecular action in situ and relate it to the structure and
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[
1987]
To my knowledge, a theory of "developmentally programmed aging" has never been explicitly stated, although the notion that aging has some relationship to development has certainly been proposed many times. In the preceding chapter (36), Dr. Hayflick has made a brief description of the central idea of developmental programming within aging. In order to discuss relevant evidence in this chapter, I would like to propose the following, somewhat more specific and operational definition: The theory of developmentally programmed aging posits that aging involves events controlled in ways recognizably similar to those that operate during development. This definition is perhaps a little less extreme than it might have been, since it uses the phrase "aging involves events" rather than the phrase "aging is caused by events." However, I think it captures most of the usual connotations of "developmentally programmed aging," and it at least has the virtue of testability. Of course, to test the theory, as defined, requires evidence of several sorts. In particular, it requires (a) that we understand how some aging events are controlled, (b) that we understand how some developmental events are controlled, and (c) that we know how to recognize whether there is or is not similarity between the two. A central message of what follows is that we are really only at the beginning of being able to test this theory, although some lines of approach do appear
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[
1981]
This chapter is in part a review of the work of others and in part a summary of recent results from our own laboratory. It attempts to cover the currently available information on apparent neurotransmitters in the small soil nematode Caenorhabditis elegans, whose advantages of genetic manipulability and cellular simplicity have recently gained it some favor in investigations of genetic control mechanisms in neural development (for review, see Riddle, 1978). Particular attention is given to mutants that affect either the level or the action of apparent neurotransmitters, since it seems likely that such mutants may have the most to offer toward the understanding of human genetic neuropathies. The general features of C. elegans are described briefly at the outset, then each apparent neurotransmitter is considered in turn, and finally a few potential implications for other organisms
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[
Methods Cell Biol,
1995]
Complementary DNA libraries are useful tools for uncovering genes of interest in C. elegans and finding specific homologies to genes in other organisms (Waterston et al., 1992; McCombie et al., 1992). When working with existing cDNA libraries, be sure to carefully choose which libraries would be most beneficial to the type of research being done. Some libraries may be specific for genes that are present in lower copy numbers, whereas others may be of a more general nature. It is important to fully understand the source and construction of the library you will be working with. Once an appropriate library has been chosen, work may begin to isolate a specific cDNA and sequence it completely or to survey many cDNAs by single-pass DNA sequencing. Whatever the project, it is important to develop a specific strategy for both the sequencing and the organization of the clones being characterized. The strategies and procedures we have outlined in this chapter have proven effective for rapid and comprehensive cDNA characterization.
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[
1977]
Radiochemical assays based on the selective extraction of either substrate or product from an aqueous reaction volume into an organic scintillator have been developed for acetylcholinesterase and choline acetyltransferase. These rapid, convenient assays have made it possible to screen large numbers of mutant lines for potential enzymatic defects. One mutant with a partial acetylcholinesterase defect and two more with choline acetyltransferase defective mutants have been identified. The acetylcholinesterase defective mutant lacks two of the four isozymic forms of acetylcholinesterase found in wild type C. elegans. Behaviorally, it is selectively defective in the propagation of contractile waves in the body region. Of the two mutants with choline acetyltransferase defects, one is remarkabley paralyzed and uncoordinated, while the other is behaviorally nearly normal.
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[
WormBook,
2005]
The mitochondrial genome is vital for Caenorhabditis elegans metabolism, physiology, and development. The C. elegans mitochondrial DNA is typical of animal mitochondrial genomes in its size and gene content. It is 13,794 nucleotides in length and encodes 36 genes: 2 ribosomal RNAs, 22 transfer RNAs, and 12 protein subunits of the mitochondrial respiratory chain. Although it represents only a small number of genes, an elaborate cellular machinery comprised of over 200 nuclear genes is needed to replicate, transcribe, and maintain the mitochondrial chromosome and to assemble the translation machinery needed to express this dozen proteins. Mitochondrial genetics is peculiar and complex because mitochondrial DNA is maternally inherited and can be present at tens to tens of thousands of copies per cell. The mitochondrial genome content of the developing nematode is developmentally regulated; it increases about 30-fold between the L1 and the adult stages and blocking the increase leads to larval arrest. Energy metabolism is also intimately linked to aging and lifespan determination. The nematode model system offers numerous advantages for understanding the full importance and scope of the mitochondrial genome in animal life.
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[
Modern Cell Biology,
1994]
During the development of any multicellular organism, the behavior of any given cell can be influenced in two ways: by its ancestry, i.e., by the particular pattern of determinants it inherits (lineal programming); or by its environment, i.e., the signals it receives from other cells. In C. elegans, the relative importance of these two factors for the development of any given cell can be examined with an unusually high degree of precision. There are a number of reasons for this, but perhaps the most important is that the cell lineage, the particular pattern of cell divisions and differentiations that occur in development, is known, and is largely the same from animal to animal. Alterations in the lineage, therefore, can be understood in terms of altered developmental decisions of