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WormBook,
2010]
Ethanol is a widely used drug whose mechanism of action, despite intensive study, remains uncertain. Biochemical and electrophysiological experiments have identified receptors and ion channels whose functions are altered at physiological concentrations of ethanol. Yet, the contribution of these potential targets to its intoxicating or behavioral effects is unclear. Unbiased forward genetic screens for resistant or hypersensitive mutants represent an attractive means of identifying the relevant molecular targets or biochemical pathways mediating the behavioral effects of neuroactive compounds. C. elegans has proven to be a particularly useful system for such studies. The behavioral effects of ethanol occur at equivalent tissue concentrations in mammals and in C. elegans, suggesting the existence of conserved drug targets in the nervous system. This chapter reviews the results of studies directed toward determining the mechanisms of action of ethanol. Studies of the neural adaptations that occur with prolonged drug exposure are also discussed. The methods used to characterize the actions of ethanol should be applicable to the characterizations of other compounds that affect the behavior of C. elegans.
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Exp Gerontol,
2005]
In many eukaryotes oxidative phosphorylation via the mitochondrial electron transport chain provides the major means of ATP production. Complete removal of this capacity often results in premature death. Recent studies using the nematode Caenorhabditis elegans are surprising because they have revealed that disruption of many of the key components of the normal mitochondrial energy-generating machinery do not result in death, rather they result in adult life span extension. Such mutants have been collectively termed Mit mutants. In this short review, the potential use of alternate metabolic pathways for energy generation by Mit mutants will be considered. The effects of using such pathways on residual mitochondrial functionality, reactive radical species production, and longevity will also be explored.
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Adv Genet,
2003]
Despite the intimate nature of the aging process we actually know little about it. In more recent years, work on a variety of organisms, utilizing approaches including demography, molecular genetics, and epidemiology, have challenged some of the more commonly held assumptions about the aging process. These studies have served to reinvigorate the field of aging research and are beginning to lead the way in a renaissance in aging research (Helfand and Inouye, 2002). Invertebrate model systems such as Drosophila and Caenorhabditis elegans that permit extensive genetic analysis are at the forefront of this renaissance.
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Methods Mol Biol,
1999]
The use of antibodies to visualize the distribution and subcellular localization of gene products powerfully complements genetic and molecular analysis of gene function in Caenorhabditis elegans. Double and triple staining protocols are particularly useful for several reasons. First, colonization of proteins either within tissues or at a subcellular level can be examined. Second, costaining with stage-specific or tissue-specific markers can define the timing and tissue specificity of antigen expression. For these types of studies it is useful to be able to collect data from multiple fluorescence wavelengths simultaneously. A confocal microscope equipped with a krypton/argon laser can simultaneously detect up to three different antigens. Using a confocal microscope it is also possible to collect a series of optical sections through a sample that allows observation of changes in distribution of the antigen in different focal planes of the tissue or cell.
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FEBS J,
2016]
In the modern research era, sequencing and high-throughput analysis have linked genetic factors with a multitude of disease states. Often times, the same cellular machinery is implicated in several different diseases and has made it challenging to drug a particular disease with minimal pleotropic consequences. It is intriguing to see how different fields of disease research can present such differing views when describing the same biological process, pathway or molecule. As observations in one field converge with research in another, we gain a more complete picture of a biological system and can accurately assess the feasibility for translational science. As an example discussed here, modulating latent stress response pathways within the cell provides exciting therapeutic potential, however, opposing views have emerged in the fields of degenerative disease and cancer. This at first glance seems logical since suppression of degenerative disease entails maintaining cell viability, while cancer aims to enhance selective senescence and cell death. Since both of these disciplines seek novel therapeutic interventions, we should not overlook how scientific biases involving one biological process may impact different disease paradigms. This article is protected by copyright. All rights reserved.
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Dev Disabil Res Rev,
2010]
The extensive conservation of mitochondrial structure, composition, and function across evolution offers a unique opportunity to expand our understanding of human mitochondrial biology and disease. By investigating the biology of much simpler model organisms, it is often possible to answer questions that are unreachable at the clinical level. Here, we review the relative utility of four different model organisms, namely the bacterium Escherichia coli, the yeast Saccharomyces cerevisiae, the nematode Caenorhabditis elegans, and the fruit fly Drosophila melanogaster, in studying the role of mitochondrial proteins relevant to human disease. E. coli are single cell, prokaryotic bacteria that have proven to be a useful model system in which to investigate mitochondrial respiratory chain protein structure and function. S. cerevisiae is a single-celled eukaryote that can grow equally well by mitochondrial-dependent respiration or by ethanol fermentation, a property that has proven to be a veritable boon for investigating mitochondrial functionality. C. elegans is a multicellular, microscopic worm that is organized into five major tissues and has proven to be a robust model animal for in vitro and in vivo studies of primary respiratory chain dysfunction and its potential therapies in humans. Studied for over a century, D. melanogaster is a classic metazoan model system offering an abundance of genetic tools and reagents that facilitates investigations of mitochondrial biology using both forward and reverse genetics. The respective strengths and limitations of each species relative to mitochondrial studies are explored. In addition, an overview is provided of major discoveries made in mitochondrial biology in each of these four model systems.
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Wiley Interdiscip Rev RNA,
2011]
Trans-splicing is the joining together of portions of two separate pre-mRNA molecules. The two distinct categories of spliceosomal trans-splicing are genic trans-splicing, which joins exons of different pre-mRNA transcripts, and spliced leader (SL) trans-splicing, which involves an exon donated from a specialized SL RNA. Both depend primarily on the same signals and components as cis-splicing. Genic trans-splicing events producing protein-coding mRNAs have been described in a variety of organisms, including Caenorhabditis elegans and Drosophila. In mammalian cells, genic trans-splicing can be associated with cancers and translocations. SL trans-splicing has mainly been studied in nematodes and trypanosomes, but there are now numerous and diverse phyla (including primitive chordates) where this type of trans-splicing has been detected. Such diversity raises questions as to the evolutionary origin of the process. Another intriguing question concerns the function of trans-splicing, as operon resolution can only account for a small proportion of the total amount of SL trans-splicing.
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Annual Review of Microbiology,
1993]
In nematodes, many mRNAs contain a common 5' terminal 22-nt sequence. This sequence, the spliced leader (SL), is acquired from a small (approximately 100 nt) SL RNA via trans-splicing. Parallel in vitro and in vivo experiments have begun to clarify both the mechanism and biological role of trans-splicing. In vitro analysis (in cell free extracts) has shown that trans-splicing is remarkably similar to the snRNP mediated removal of intervening sequences from pre-mRNAs (cis-splicing). Additionally, this analysis has suggested a mechanism that may explain how the two substrates of trans-splicing (the SL RNA and pre-mRNA) efficiently associate with one another in the absence of sequence complementarity. In vivo experiments suggest that a major biological function of trans-splicing in nematodes may be to process polycistronic transcription units. Results obtained from the study of both parasitic and free-living species are discussed, and trans-splicing in nematodes is compared and contrasted to the analogous process in trypanosomatid protozoans.
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J Muscle Res Cell Motil,
2002]
Elastic proteins in the muscles of a nematode (Caenorhabditis elegans), three insects (Drosophila melanogaster, Anopheles gambiae, Bombyx mori) and a crustacean (Procambus clarkii) were compared. The sequences of thick filament proteins, twitchin in the worm and projectin in the insects, have repeating modules with fibronectin-like (Fn) and immunoglobulin-like (Ig) domains conserved between species. Projectin has additional tandem Igs and an elastic PEVK domain near the N-terminus. All the species have a second elastic protein we have called SLS protein after the Drosophila gene, sallimus. SLS protein is in the I-band. The N-terminal region has the sequence of kettin which is a spliced product of the gene composed of Ig-linker modules binding to actin. Downstream of kettin, SLS protein has two PEVK domains, unique sequence, tandem Igs, and Fn domains at the end. PEVK domains have repeating sequences: some are long and highly conserved and would have varying elasticity appropriate to different muscles. Insect indirect flight muscle (IFM) has short I-bands and electron micrographs of Lethocerus IFM show fine filaments branching from the end of thick filaments to join thin filaments before they enter the Z-disc. Projectin and kettin are in this region and the contribution of these to the high passive stiffness of Drosophila IFM myofibrils was measured from the force response to length oscillations. Kettin is attached both to actin near the Z-disc and to the end of thick filaments, and extraction of actin or digestion of kettin leads to rapid decrease in stiffness; residual tension is attributable to projectin. The wormlike chain model for polymer elasticity fitted the force-extension curve of IFM myofibrils and the number of predicted Igs in the chain is consistent with the tandem Igs in Drosophila SLS protein. We conclude that passive tension is due to kettin and projectin, either separate or
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[
BioEssays,
1993]
In trans-splicing, the pre-mRNA products of two different genes are spliced together to form a single, mature mRNA. In one type of trans-splicing, pre-mRNAs of many different genes receive a single, short leader, called spliced leader or SL. This type of trans-splicing was first discovered in the primitive eukaryotes, the trypanosomes, where it is apparently the only kind of nuclear mRNA splicing. Subsequently, it was discovered in nematodes (round worms), trematodes (flat worms), and euglena. Although this type of trans-splicing has never been found in any of the other well-studied organisms, Bruzik and Maniatis have recently reported that mammalian cells are capable of performing the reaction when they are provided with the appropriate pre-mRNAs.