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[
WormBook,
2006]
Many pathogens that can infect C. elegans have been described, including some that co-exist with the nematode in its natural environment. This chapter describes our current understanding of the different innate immune responses of C. elegans that follow infection. It focuses on the main signalling pathways that have been identified and highlights the inclusion of certain molecular cassettes in both immune and developmental functions.
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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,
2005]
About 70% of C. elegans mRNAs are trans-spliced to one of two 22 nucleotide spliced leaders. SL1 is used to trim off the 5'' ends of pre-mRNAs and replace them with the SL1 sequence. This processing event is very closely related to cis-splicing, or intron removal. The SL1 sequence is donated by a 100 nt small nuclear ribonucleoprotein particle (snRNP). This snRNP is structurally and functionally related to the U snRNAs (U1, U2, U4, U5 and U6) that play key roles in intron removal and trans-splicing, except that it is consumed in the process of splicing. More than half of C. elegans pre-mRNAs are subject to SL1 trans-splicing. About 30% are not trans-spliced at all. The remaining genes are trans-spliced by SL2. These genes are all downstream genes in closely spaced gene clusters similar to bacterial operons. They are transcribed from a promoter at the 5'' end of the cluster of between 2 and 8 genes. This transcription makes a polycistronic pre-mRNA that is co-transcriptionally processed by cleavage and polyadenylation at the 3'' end of each gene, and this event is closely coupled to the SL2 trans-splicing event that occurs only ~100 nt further downstream. Recent studies on the mechanism of SL2 trans-splicing have revealed that one of the 3'' end formation proteins, CstF, interacts with the only protein known to be specific to the SL2 snRNP. The operons contain primarily genes whose products are needed for mitochondrial function and the basic machinery of gene expression: transcription, splicing and translation. Many operons contain genes whose products are known to function together. This presumably provides co-regulation of these proteins by producing a single RNA that encodes both.
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[
WormBook,
2005]
TGF-beta superfamily ligands play fundamental roles in the development and physiology of diverse animal species. Genetic and genomic analyses in the model organism Caenorhabditis elegans have contributed to the understanding of TGF-beta -related signal transduction mechanisms. In this chapter, I describe the currently characterized TGF-beta -related signals and signal transduction cassettes in C. elegans. Homology searches of the genome identify five TGF-beta -related genes, for which functions have been identified for three. Two of the TGF-beta -related genes,
daf-7 and
dbl-1 , function through conventional signaling pathways. These signaling pathways are comprised of ser/thr kinase receptors, Smads, and transcription co-factors. A third TGF-beta -related gene,
unc-129 , functions in axonal guidance using novel signaling mechanisms. Thus, TGF-beta -related signaling in C. elegans proceeds via both conserved and novel paradigms that can inform studies in other animal systems.
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[
WormBook,
2007]
The nematode cuticle is an extremely flexible and resilient exoskeleton that permits locomotion via attachment to muscle, confers environmental protection and allows growth by molting. It is synthesised five times, once in the embryo and subsequently at the end of each larval stage prior to molting. It is a highly structured extra-cellular matrix (ECM), composed predominantly of cross-linked collagens, additional insoluble proteins termed cuticlins, associated glycoproteins and lipids. The cuticle collagens are encoded by a large gene family that are subject to strict patterns of temporal regulation. Cuticle collagen biosynthesis involves numerous co- and post-translational modification, processing, secretion and cross-linking steps that in turn are catalysed by specific enzymes and chaperones. Mutations in individual collagen genes and their biosynthetic pathway components can result in a range of defects from abnormal morphology (dumpy and blister) to embryonic and larval death, confirming an essential role for this structure and highlighting its potential as an ECM experimental model system.
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[
1994]
The current interest in the nematode Caenorhabditis elegans began approximately 25 years ago when Sidney Brenner selected this species as the most suitable for studies of metazoan development and nervous system. The basis of this selection rested on the anatomical simplicity of nematodes, which nevertheless possess the major differentiated cell types of higher animals, and the tractability of C. elegans to the genetic approach. Over the past two decades or so, progress has been impressive: the cell lineage from egg to adult and the anatomy of the nervous system have been completely described, genetic investigations of numerous developmental problems are co-ordinated within a universally-agreed, systematic nomenclature, a physical map of the C. elegans genome is nearing completion and a project to sequence the entire genome is underway. Furthermore, the number of laboratories seeking to understand the mechanisms controlling animal development through genetic and molecular investigations of C. elegans is rising rapidly as the advantages of this organism become
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[
1980]
The free-living nematode Caenorhabditis elegans has attracted attention in recent years as an organism for the study of the genetic control of development. This chapter briefly describes the present state of this work. Many of the studies reported on here have not yet been published but have been described in "The Worm Breeder's Gazette", an informal newsletter I edit, and at a C. elegans meeting held at Cold Spring Harbor in May 1979. A previous review of this field was written by Riddle (1978). The use of free-living nematodes in genetic studies was first suggested by Dougherty and Calhoun in 1948. Early studies of C. elegans by Dougherty and co-workers (1959) emphasized methods of axenic cultivation while the sexual cycle was described by Nigon (1949). The present interest in C. elegans, however, was triggered by Sydney Brenner who took up the organism in the late 1960s as a possibly useful organism for the study of the genetic control of the nervous system and of behavior (Brenner, 1973). It was largely due to Brenner (1974) that the present methods of cultivation and of genetic analysis were developed.
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[
WormBook,
2006]
In Drosophila and vertebrates, Hedgehog (Hh) signalling is mediated by a cascade of genes, which play essential roles in cell proliferation and survival, and in patterning of the embryo, limb buds and organs. In C. elegans, this pathway has undergone considerable evolutionary divergence; genes encoding homologues of key pathway members, including Hh, Smoothened, Cos2, Fused and Suppressor of Fused, are absent. Surprisingly, over sixty proteins (i.e. WRT, GRD, GRL, and QUA), encoded by a set of genes collectively referred to as the Hh-related genes, and two co-orthologs ( PTC-1 ,-3) of fly Patched, a Hh receptor, are present in C. elegans. Several of the Hh-related proteins are bipartite and all can potentially generate peptides with signalling activity, although none of these peptides shares obvious sequence similarity with Hh. In addition, the ptc -related ( ptr ) genes, which are present in a single copy in Drosophila and vertebrates and encode proteins closely related to Patched, have undergone an expansion in number in nematodes. A number of functions, including roles in molting, have been attributed to the C. elegans Hh-related, PTC and PTR proteins; most of these functions involve processes that are associated with the trafficking of proteins, sterols or sterol-modified proteins. Genes encoding other components of the Hh signalling pathway are also found in C. elegans, but their functions remain to be elucidated.
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[
1994]
Nematodes have been cultured continuously in the laboratory since 1944 when Margaret Briggs Gochnauer isolated and cultured the free-living hermaphroditic species Caenorhabditis briggsae. Work with C. briggsae and other rhabditid nematodes, C. elegans, Rhabditis anomala, and R. pellio, demonstrated the relative ease with which they could be cultured. The culturing techniques described here were developed for C. elegans, but are generally suitable (to varying degrees) for other free-living nematodes. Whereas much of the early work involved axenic culturing, most of these techniques are no longer in common use and are not included here. In the 1970s C. elegans became the predominant research model due to work by Brenner and co-workers on the genetics and development of this species. An adult C. elegans is about 1.5 mm long, and under optimal laboratory conditions has a life cycle of approximately 3 days. There are two sexes, males and self-fertile hermaphrodites, that are readily distinguishable as adults. The animals are transparent throughout the life cycle, permitting observation of cell divisions in living animals using differential interference microscopy. The complete cell lineage and neural circuitry have been determined and a large collection of behavioral and anatomical mutants have been isolated. C. elegans has six developmental stages: egg, four larval stages (L1-L4), and adult. Under starvation conditions or specific manipulations of the culture conditions a developmentally arrested dispersal stage, the dauer larva, can be formed as an alternative third larval stage. Many of the protocols included here and other experimental protocols have been summarized in "The Nematode Caenorhabditis elegans". We also include a previously unpublished method for long-term chemostat cultures of C. elegans. General laboratory culture conditions for nematode parasites of animals have been described, but none of these nematodes can be cultured in the laboratory through more than one life cycle. Marine nematodes and some plant parasites have been cultured xenically or with fungi. Laboratory cultivation of several plant parasites on Arabidopsis thaliana seedlings in agar petri plates has also been reported.