Mouchiroud L et al. (2016) Curr Protoc Neurosci "The Movement Tracker: A Flexible System for Automated Movement Analysis in Invertebrate Model Organisms."

Sorrentino V, Deplancke B, Ouhmad T, Cornaglia M, Gijs MA, Auwerx J, Williams EG, Krishnamani G, Frochaux MV, Nicolet-Dit-Felix AA, Lin T, Mouchiroud L

[Curr Protoc Neurosci, 2016]

Phenotyping strategies in simple model organisms such as D. melanogaster and C. elegans are often broadly limited to growth, aging, and fitness. Recently, a number of physical setups and video tracking software suites have been developed to allow for accurate, quantitative, and high-throughput analysis of movement in flies and worms. However, many of these systems require precise experimental setups and/or fixed recording formats. We report here an update to the Parallel Worm Tracker software, which we termed the Movement Tracker. The Movement Tracker allows variable experimental setups to provide cross-platform automated processing of a variety of movement characteristics in both worms and flies and permits the use of simple physical setups that can be readily implemented in any laboratory. This software allows high-throughput processing capabilities and high levels of flexibility in video analysis, providing quantitative movement data on C. elegans and D. melanogaster in a variety of different conditions. 2016 by John Wiley and Sons, Inc.

Popiel E et al. (2020) Methods Mol Biol "Generation and Analysis of CCM Phenotypes in C. elegans"

Popiel E, Derry WB

[Methods Mol Biol, 2020]

This chapter presents methods for exploiting the powerful tools available in the nematode worm Caenorhabditis elegans to understand the in vivo functions of cerebral cavernous malformation (CCM) genes and the organization of their associated signaling pathways. Included are methods for assessing phenotypes caused by loss-of-function mutations in the worm CCM genes kri-1 and ccm-3, CRISPR-based gene editing techniques, and protocols for conducting high-throughput forward genetic and small molecule screens.

Mishur RJ et al. (2013) Methods Mol Biol "Exometabolomic mapping of Caenorhabditis elegans: a tool to noninvasively investigate aging."

Rea SL, Mishur RJ, Butler JA

[Methods Mol Biol, 2013]

Metabolomic analyses can provide valuable information about the internal metabolism of an organism; however, these studies can become quickly complicated by the large number of metabolites that are often detected. Overcoming this limitation requires high-resolution analytical separation techniques, coupled with high-power deconvolution software. Additionally, much care must be taken in metabolomic sample preparation to quench active enzymes and avoid artifactual changes in the metabolome. Here we present a relatively simple and straightforward technique, exometabolome mapping, which bypasses each of these concerns, is noninvasive, and provides a concise summary of the key metabolic processes operative in an organism. We illustrate our method using the nematode C. elegans, an organism which has been widely exploited in aging studies; however, with only minimal modification, our technique is extendible to other sample types, and indeed we have successfully used it both to perform yeast footprinting and to study the excreted metabolic end products of human kidney cancer cell lines.

Jeremy Nance et al. (2005) WormBook "Gastrulation in C. elegans."

Bob Goldstein, Jeremy Nance, Jen-Yi Lee

[WormBook, 2005]

Gastrulation is the process by which the germ layers become positioned in an embryo. C. elegans gastrulation serves as a model for studying the molecular mechanisms of diverse cellular and developmental phenomena, including morphogenesis, cell polarization, cell-cell signaling, actomyosin contraction and cell-cell adhesion. One distinct advantage of studying these phenomena in C. elegans is that genetic tools can be combined with high resolution live cell imaging and direct manipulations of the cells involved. Here we review what is known to date about the cellular and molecular mechanisms that function in C. elegans gastrulation.

Ciche T (2007) WormBook "The biology and genome of Heterorhabditis bacteriophora."

Ciche T

[WormBook, 2007]

Heterorhabditis bacteriophora is an entomopathogenic nematode (EPN) mutually associated with the enteric bacterium, Photorhabdus luminescens, used globally for the biological control of insects. Much of the previous research concerning H. bacteriophora has dealt with applied aspects related to biological control. However, H. bacteriophora is an excellent model to investigate fundamental processes such as parasitism and mutualism in addition to its comparative value to Caenorhabditis elegans. In June 2005, H. bacteriophora was targeted by NHGRI for a high quality genome sequence. This chapter summarizes the biology of H. bacteriophora in common and distinct from C. elegans, as well as the status of the genome project.

Favello A et al. (1995) Methods Cell Biol "Genomic DNA sequencing methods."

Wilson RK, Hillier L, Favello A

[Methods Cell Biol, 1995]

Sequence analysis of cosmids from C. elegans and other organisms currently is best done using the random or "shotgun" strategy (Wilson et al., 1994). After shearing by sonication, DNA is used to prepare M13 subclone libraries which provide good coverage and high-quality sequence data. The subclones are assembled and the data edited using software tools developed especially for C. elegans genomic sequencing. These same tools facilitate much of the subsequent work to complete both strands of the sequence and resolve any remaining ambiguities. Analysis of the finished sequence is then accomplished using several additional computer tools including Genefinder and ACeDB. Taken together, these methods and tools provide a powerful means for genome analysis in the nematode.

Edgley ML et al. (1995) Methods Cell Biol "Genetic balancers."

Riddle DL, Baillie DL, Edgley ML, Rose AM

[Methods Cell Biol, 1995]

Genetic balancers are genetic constructs or chromosomal rearrangements that allow lethal or sterile mutations to be stably maintained in heterozygotes. In this chapter we use the term balancer primarily to refer to chromosomal duplications or rearrangements that suppress crossing over. In addition, we define lethal as any mutation that blocks survival or reproduction. Phenotypes associated with lethal mutations in Caenorhabditis elegans range from egg or larval lethality to adult sterility and maternal effect lethality, and can include conditional effects such as temperature sensitivity. The number of essential genes in C. elegans (those identified by lethal mutations) may range as high as 7000 according to genetic estimates. Thus, lethal mutations constitute a rich source of information about basic biological processes in this nematode.

Valerie Reinke (2006) WormBook "Germline genomics."

Valerie Reinke

[WormBook, 2006]

The completion of the C. elegans genome sequence permits the comprehensive examination of the expression and function of genes. Annotation of virtually every encoded gene in the genome allows systematic analysis of those genes using high-throughput assays, such as microarrays and RNAi. This chapter will center on the use of microarrays to comprehensively identify genes with enriched expression in the germ line during development. This knowledge provides a database for further studies that focus on gene function during germline development or early embryogenesis. Additionally, a comprehensive overview of germline gene expression can uncover striking biases in how genes expressed in the germ line are distributed in the genome, leading to new discoveries of global regulatory mechanisms in the germ line.

Tuck SP et al. (1994) Modern Cell Biology "Cell interactions and signal transduction in C. elegans development."

Greenwald IS, Tuck SP

[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

Green RA et al. (2008) Methods Cell Biol "Expression and imaging of fluorescent proteins in the C. elegans gonad and early embryo."

Portier N, Monen J, Dammermann A, Pemble H, Hyman A, Desai A, Audhya A, Green RA, Oegema K, Pozniakovsky A

[Methods Cell Biol, 2008]

The Caenorhabditis elegans gonad and early embryo have recently emerged as an attractive metazoan model system for studying cell and developmental biology. The success of this system is attributable to the stereotypical architecture and reproducible cell divisions of the gonad/early embryo, coupled with penetrant RNAi-mediated protein depletion. These features have facilitated the development of visual assays with high spatiotemporal resolution to monitor specific subcellular processes. Assay development has relied heavily on the emergence of methods to circumvent germline silencing to allow the expression of transgenes encoding fluorescent fusion proteins. In this chapter, we discuss methods for the expression and imaging of fluorescent proteins in the C. elegans germline, including the design of transgenes for optimal expression, the generation of transgenic worm lines by ballistic bombardment, the construction of multimarker lines by mating, and methods for live imaging of the gonad and early embryo.