Role of autophagy in Caenorhabditis elegans

Autophagy is an evolutionarily conserved intracellular catabolic system. During Caenorhabditis elegans development, autophagy plays an important role in many physiological processes, including survival under starvation conditions, modulation of life span, and regulation of necrotic cell death caused by toxic ion-channel variants. Recently, it has been demonstrated that during embryogenesis, basal levels of autophagy selectively remove a group of proteins in somatic cells, including the aggregate-prone components of germline P granules. Degradation of these protein aggregates provides a genetic model to identify essential autophagy components and also to elucidate how the autophagic machinery selectively recognizes and degrades specific targets during animal development.     

基于模式生物秀丽隐杆线虫的三丁基锡生态毒性评价

本研究利用秀丽隐杆线虫半致死浓度分析和致死曲线分析来筛选对三丁基锡（Tributyltin,TBT）敏感的线虫品系,并讨论与TBT毒理学过程可能相关的基因。通过对秀丽隐杆线虫体长、每窝子代数和怀卵量的测定来探讨TBT的生态毒性效应,以期为TBT对秀丽隐杆线虫和人类的生态毒性评价和致毒机理研究提供科学依据。结果表明：TBT对各品系线虫48 h LC50从小到大依次为egl-1（n487）＜ced-4（n1162）＜cep-1（gk138）=cep-1（lg12501）＜ced-9（n1950）＜clk-2（mn159）＜ced-3（n717）＜N2＜opIs34（hus-1::GFP） ＜opIs56（egl-1::GFP）＜daf-16（mn86）＜hus-1（op241）＜daf-2（e1370）。本研究筛选出对TBT最敏感的线虫品系为egl-1（n487）,而对TBT耐受力最强的是daf-2（e1370）。TBT对秀丽隐杆线虫体长、每窝子代数和怀卵量均呈现浓度依赖型的抑制作用。     

Functional characterisation of a cyst nematode acetylcholinesterase gene using Caenorhabditis elegans as a heterologous system

Migration of plant-parasitic nematode infective larval stages through soil and invasion of roots requires perception and integration of sensory cues culminating in particular responses that lead to root penetration and parasite establishment. Components of the chemoreceptive neuronal circuitry involved in these responses are targets for control measures aimed at preventing infection. Here we report, to our knowledge, the first isolation of cyst nematode ace-2 genes encoding acetylcholinesterase (AChE). The ace-2 genes from Globodera pallida (Gp-ace-2) and Heterodera glycines (Hg-ace-2) show homology to ace-2 of Caenorhabditis elegans (Ce-ace-2). Gp-ace-2 is expressed most highly in the infective J2 stage with lowest expression in the early parasitic stages. Expression and functional analysis of the Globodera gene were carried out using the free-living nematode C. elegans in order to overcome the refractory nature of the obligate parasite G. pallida to many biological studies. Caenorhabditis elegans transformed with a GFP reporter construct under the control of the Gp-ace-2 promoter exhibited specific and restricted GFP expression in neuronal cells in the head ganglia. Gp-ACE-2 protein can functionally complement its C. elegans homologue. A chimeric construct containing the Ce-ace-2 promoter region and the Gp-ace-2 coding region and 3′ untranslated region was able to restore a normal phenotype to the uncoordinated C. elegans double mutant ace-1;ace-2. This study demonstrates conservation of AChE function and expression between free-living and plant-parasitic nematode species, and highlights the utility of C. elegans as a heterologous system to study neuronal aspects of plant-parasitic nematode biology.     

Functional assessment of Nramp-like metal transporters and manganese in Caenorhabditis elegans

Nramp1 (natural resistance-associated macrophage protein-1) is a functionally conserved iron-manganese transporter in macrophages. Manganese (Mn), a superoxide scavenger, is required in trace amounts and functions as a cofactor for most antioxidants. Three Nramp homologs, smf-1, smf-2, and smf-3, have been identified thus far in the nematode Caenorhabditis elegans. A GFP promoter assay revealed largely intestinal expression of the smf genes from early embryonic through adult stages. In addition, smf deletion mutants showed increased sensitivity to excess Mn and mild sensitivity to EDTA. Interestingly, these smf deletion mutants demonstrated hypersensitivity to the pathogen Staphylococcus aureus, an effect that was rescued by Mn feeding or knockdown of the Golgi calcium/manganese ATPase, pmr-1, indicating that Mn uptake is essential for the innate immune system. This reversal of pathogen sensitivity by Mn feeding suggests a protective and therapeutic role of Mn in pathogen evasion systems. We propose that the C. elegans intestinal lumen may mimic the mammalian macrophage phagosome and thus could be a simple model for studying Mn-mediated innate immunity.     

秀丽隐杆线虫脂肪沉积研究进展

脂肪的过度沉积会引发多种疾病,如心脏病、高血压、高甘油三酯血症、Ⅱ型糖尿病等。小白鼠(Mus musculus)和猪(Sus domesticus)是常用的研究脂肪沉积的模式动物,近年来随着研究的深入,发现脂肪代谢调控网络错综复杂,调控因子相互作用。秀丽隐杆线虫(Caenorhabditis elegans)具有结构简单、身体透明、便于观察、繁殖周期短、易于人工培养等特征,因此使得秀丽隐杆线虫进行脂肪调控的研究成为了可能。本文通过总结国内外线虫脂肪沉积方面的研究,综述秀丽隐杆线虫研究脂肪沉积的进展。     

Ce-wts-1 plays important roles in Caenorhabditis elegans development

The Hippo-Warts pathway defines a novel signaling cascade involved in organ size control and tumor suppression. However, the developmental function of this pathway is less understood. Here we report that the Caenorhabditis elegans homolog of Warts, Ce-wts-1, plays important roles during worm development. The null allele of Ce-wts-1 causes L1 lethality. Partial loss of Ce-wts-1 function by RNAi reveals that Ce-wts-1 is involved in many developmental processes such as larval development, growth rate regulation, gut granule formation, pharynx development, dauer formation, lifespan and body length control. Genetic analyses show that Ce-wts-1 functions synergistically with the TGF-β Sma/Mab pathway to regulate body length. In addition, CE-WTS-1::GFP is enriched near the inner cell membrane, implying its possible membrane-related function.     

Systematic comparison of bacterial feeding strains for increased yield of Caenorhabditis elegans males by RNA interference-induced non-disjunction

Rare Caenorhabditis elegans males arise when sex chromosome non-disjunction occurs during meiosis in self-fertilizing hermaphrodites. Non-disjunction is a relatively rare event, and males are typically observed at a frequency of less than one in five hundred wild-type animals. Males are required for genetic crosses and phenotypic analysis, yet current methods to generate large numbers of males can be cumbersome. Here, we identify RNAi reagents (dsRNA-expressing bacteria) with improved effectiveness for eliciting males. Specifically, we used RNAi to systematically reduce the expression of over two hundred genes with meiotic chromosome segregation functions, and we identified a set of RNAi reagents that robustly and reproducibly elicited male progeny.     

Nanoparticle imaging and diagnostic of Caenorhabditis elegans intracellular pH

A novel diagnostic tool has been developed for the characterization of intracellular pH (pHi) in the model organism Caenorhabditis elegans. This tool exploits the chemical stability of colloidal silica and the pH sensitivity of certain fluorescent dyes. Once ingested, the fluorescent colloidal dispersion yields a reliable visual indication of pH without the use of chemical fixatives or damaging the nematode. The pH-sensitive silica nanoparticles were visualized by confocal microscopy, and the fluorescence spectra from the internally referenced colloidal particulates were measured. By comparing the fluorescence profile of colloids in wild-type (N2) and mutant (eat-3) C. elegans against a calibration series, the intestinal pHi could be established in each population. The rapid characterization of pHi using this inexpensive nonintrusive technique has significant implications for disease research where C. elegans is used as a model organism.     

Apigenin inhibits larval growth of Caenorhabditis elegans through DAF-16 activation

Treatment of Caenorhabditis elegans with apigenin, 5,7,4′-trihydroxyflavone, induces larval growth inhibition. To understand the molecular basis of apigenin-induced larval growth inhibition, the effects of apigenin on DAF-16 activity were examined. DAF-16 was activated through nuclear translocation and the mRNA level of sod-3, one of the known DAF-16 target genes, was increased upon apigenin treatment. DAF-16 activity was required for the growth inhibition, since the larval growth retardation upon apigenin treatment was suppressed in daf-16 mutants. These results indicate that apigenin acts as a stressor that activates DAF-16, which in turn inhibits larval growth.     

Medicinal plants and antioxidants: What do we learn from cell culture and Caenorhabditis elegans studies?

Traditional medicinal plants have a long history of therapeutic use. The beneficial health effects of medicinal plants rich in polyphenols are often attributed to their potent antioxidant activities, as established in vitro, since diets rich in polyphenols are epidemiologically associated with a decreased incidence of age-related diseases in humans. However, medicinal plants may also exert pro-oxidant effects that up-regulate endogenous protective enzymes. Care is needed when studying the biological effects of medicinal plants in cell culture because some polyphenols oxidize readily in culture media. This review summarizes the data we have obtained from in vitro and in vivo (Caenorhabditis elegans) studies examining the diverse effects of traditional medicinal plants and their modes of action.     

Spatio-temporal reference model of Caenorhabditis elegans embryogenesis with cell contact maps

The nematode Caenorhabditis elegans has been used as a model for developmental biology for decades. Still, the few publicly available spatio-temporal (4D) data sets have conflicting information regarding variability of cell positions and are not well-suited for a standard 4D embryonic model, due to compression. We have recorded six uncompressed embryos, and determined their lineage and 4D coordinates, including nuclear radii, until the end of gastrulation. We find a remarkable degree of stability in the cell positions, as well as little rotational movement, which allowed us to combine the data into a single reference model of C. elegans embryogenesis. Using Voronoi decomposition we generated the list of all predicted cell contacts during early embryogenesis and calculated these contacts up to the ∼ 150 cell stage, and find that about 1500 contacts last 2.5 min or longer. The cell contact map allows for comparison of multiple 4D data sets, e.g., mutants or related species, at the cellular level. A comparison of our uncompressed 4D model with a compressed embryo shows that up to 40% of the cell contacts can be different. To visualize the 4D model interactively we developed a software utility. Our model provides an anatomical resource and can serve as foundation to display 4D expression data, a basis for developmental systems biology.     

Cholesterol and the biosynthesis of glycosphingolipids are required for sperm activation in Caenorhabditis elegans

Ejaculated mammalian sperm must acquire fertilization capacity after residing into the female reproductive tract, a process collectively known as capacitation. Cholesterol efflux was required for sperm maturation. Different from flagellated sperm, C. elegans sperm are crawling cells. C. elegans sperm are highly enriched with cholesterol though this animal species lacks biosynthetic pathway for cholesterol and its survival requires an exogenous cholesterol supply. The low abundance of cholesterol in C. elegans lipid extract is thought insufficient to form lipid microdomains ubiquitously in this organism. We present evidence that cholesterol is enriched in the plasma membrane of C. elegans spermatids and that cholesterol- and glycosphingolipids (GSLs)-enriched membrane microdomains (lipid microdomains) mediate sperm activation. Disruption of sperm lipid microdomains by acute manipulation of cholesterol in vitro blocks the sperm activation. Restriction of cholesterol uptake also results in the abnormal sperm activation in both males and hermaphrodites. Manipulation of the integrity of lipid microdomains by targeting the biosynthesis of GSLs inhibits sperm activation and the inhibition can be rescued by the addition of exogenous GSLs. The cleavage of glycosylphosphatidylinositol (GPI)-anchored proteins, which are exclusively found in lipid microdomains, also affects sperm activation. We conclude that localized signaling mediated by lipid microdomains is critical for worm sperm activation. Lipid microdomains composed of cholesterol and GSLs have been observed in flagellated sperm of several animal species, thus cholesterol, before its efflux from the plasma membrane, might be needed to assemble into a platform for some more important upstream signal sorting during spermatogenesis than was previously thought.     

The olfactory signal transduction for attractive odorants in Caenorhabditis elegans

Olfaction in Caenorhabditis elegans is a versatile and sensitive strategy to seek food and avoid danger by sensing volatile chemicals emitted by the targets. The ability to sense attractive odor is mainly accomplished by the AWA and AWC neurons. Previous studies have shown the components of the olfaction signal pathway in these two amphid chemosensory neurons, but integration of the individual signaling components requires further elucidation. Here we review the progresses in our understanding of signal pathways for attractive olfaction involving AWA and AWC neurons, and discuss how the different signal molecules might employ the common molecular cascades to transduce the olfactory system and guide behavior in each neuron.     

Deceptively simple but simply deceptive - Caenorhabditis elegans lifespan studies: Considerations for aging and antioxidant effects

The nematode worm Caenorhabditis elegans (C. elegans) is increasingly popular as a model organism for aging studies as well as for testing antioxidants and other compounds for effects on longevity. However, results in the literature are sometimes confusing and contradictory [1], [2], [3] and [4]. This review introduces C. elegans as a model organism, discusses aspects that make it attractive for aging and antioxidant research, and addresses some problems and potential artifacts.     

Solution structure of CEH-37 homeodomain of the nematode Caenorhabditis elegans

The nematode Caenorhabditis elegans protein CEH-37 belongs to the paired OTD/OTX family of homeobox-containing homeodomain proteins. CEH-37 shares sequence similarity with homeodomain proteins, although it specifically binds to double-stranded C. elegans telomeric DNA, which is unusual to homeodomain proteins. Here, we report the solution structure of CEH-37 homeodomain and molecular interaction with double-stranded C. elegans telomeric DNA using nuclear magnetic resonance (NMR) spectroscopy. NMR structure shows that CEH-37 homeodomain is composed of a flexible N-terminal region and three α-helices with a helix-turn-helix (HTH) DNA binding motif. Data from size-exclusion chromatography and fluorescence spectroscopy reveal that CEH-37 homeodomain interacts strongly with double-stranded C. elegans telomeric DNA. NMR titration experiments identified residues responsible for specific binding to nematode double-stranded telomeric DNA. These results suggest that C. elegans homeodomain protein, CEH-37 could play an important role in telomere function via DNA binding.