Localization of FMRFamide-like peptides in Caenorhabditis elegans.

The neuropeptide FMRFamide (Phe-Met-Arg-Phe-NH2) is a member of a large family of related peptides that have been found throughout the animal kingdom. By using an antiserum specific for the Arg-Phe-NH2 moiety, we have found that about 10% of the neurons in the nematode Caenorhabditis elegans are immunoreactive. Most of these neurons, which include sensory, motor, and interneurons, were identified on the basis of their number, position, and projection pattern and by analysis of characterized mutants. Neurons that were immunoreactive in hermaphrodite animals were generally also found in males, but each sex had, in addition, sex-specific immunoreactive cells. Staining of hermaphrodite animals from different larval stages suggests that the onset of FMRFamide-like expression is differentially regulated among the cells. We have found a possible neuromodulatory role for the related peptide FLRFamide (Phe-Leu-Arg-Phe-NH2). In an egg-laying assay, FLRFamide by itself was not active but could potentiate a serotonin effect. The FMRFamide-like immunoreactivity was also used as a marker to examine the differentiation of cells that normally undergo programmed cell death. Cells that are destined to die in the Pn.a lineages appear to differentiate and adopt the fate of lineally equivalent cells before cell death.     

FMRFamide-related neuropeptide gene family in Caenorhabditis elegans

Neuropeptides are used as signaling molecules in the nervous system of most organisms, including mammals. The family of FMRFamide (Phe-Met-Arg-Phe-NH2)-like neuropeptides (FaRPs) all share an RFamide sequence at their C-termini and have been shown to have diverse functions in the central and peripheral nervous systems. In the nematode Caenorhabditis elegans, FMRFamide-like peptides (FaRPs) are expressed in at least 10% of the neurons, including motor, sensory, and interneurons that are involved in movement, feeding, defecation, and reproduction. Twenty-two genes, designated flp-1 through flp-22, encode FaRPs in C. elegans, although there are likely to be additional flp genes to be identified. Each flp gene encodes a different set of FaRPs, yielding a predicted total of 59 distinct FaRPs; a few of the genes may also encode non-FaRPs. Inactivation of some of the flp genes indicates that at least one flp gene has unique functions, while at least two flp genes appear to have overlapping functions with other flp genes. These results suggest that a complex family of FaRPs have varied roles through all stages of development and in adulthood in C. elegans.     

Dopaminergic neurons in the nematode Caenorhabditis elegans.

Dopamine is the putative transmitter of eight neurons in the hermaphrodite form of the nematode Caenorhabditis elegans. These include the cephalic and deirid neurons, which are believed to be mechanosensory. The male has an additional six dopaminergic neurons in the tail. Mutants have been selected which have defects in the formaldehyde induced fluorescence and lack dopamine to varying degrees, but they are not insensitive to touch. The dopaminergic neurons of C. elegans are compared with the homologous neurons in Ascaris lumbricoides.     

Multiple ace genes encoding acetylcholinesterases of Caenorhabditis elegans have distinct tissue expression

ace-1 and ace-2 genes encoding acetylcholinesterase in the nematode Caenorhabditis elegans present 35% identity in coding sequences but no homology in noncoding regions (introns, 5'- and 3'-untranslated regions). A 5'-region of ace-2 was defined by rescue of ace-1;ace-2 mutants. When green fluorescent protein (GFP) expression was driven by this regulatory region, the resulting pattern was distinct from that of ace-1. This latter gene is expressed in all body-wall and vulval muscle cells (Culetto et al., 1999), whereas ace-2 is expressed almost exclusively in neurons. ace-3 and ace-4 genes are located in close proximity on chromosome II (Combes et al., 2000). These two genes were first transcribed in vivo as a bicistronic messenger and thus constitute an ace-3;ace-4 operon. However, there was a very low level of monocistronic mRNA of ace-4 (the upstream gene) in vivo, and no ACE-4 enzymatic activity was ever detected. GFP expression driven by a 5' upstream region of the ace-3;ace-4 operon was detected in several muscle cells of the pharynx (pm3, pm4, pm5 and pm7) and in the two canal associated neurons (CAN cells). A dorsal row of body-wall muscle cells was intensively labelled in larval stages but no longer detected in adults. The distinct tissue-specific expression of ace-1, ace-2 and ace-3 (coexpressed only in pm5 cells) indicates that ace genes are not redundant.     

Expression and regulation of an FMRFamide-related neuropeptide gene family in Caenorhabditis elegans

FMRFamide (Phe-Met-Arg-Phe-NH2) and related peptides (FaRPs) have been found throughout the animal kingdom, where they are involved in many behaviors. We previously identified 22 genes comprising the flp gene family that encodes FaRPs in Caenorhabditis elegans; in this paper we report the identification of another flp gene, flp-23. As a first step toward determining their functional roles in C. elegans, we examined the cell-specific expression pattern of the flp gene family. Of the 19 flp genes examined, each gene is expressed in a distinct set of cells; these cells include interneurons, motor neurons, and sensory neurons that are involved in multiple behaviors, as well as supporting cells, muscle cells, and epidermal cells. Several flp genes show sex-specific expression patterns. Furthermore, we find that expression of two flp genes changes in response to the developmental state of the animal. Many neurons express multiple flp genes. To investigate how flp genes are regulated in different neuronal subtypes, we examined flp expression in a small, well-defined subset of neurons, the mechanosensory neurons. Mutations in the unc-86 and mec-3 genes, which are necessary for the production and differentiation of the mechanosensory neurons, result in the complete loss of flp-4, flp-8, and flp-20 expression in mechanosensory neurons. Collectively, these data indicate that members of the flp gene family are likely to influence multiple behaviors and that their regulation can be dependent on the developmental state of the organism.     

SCPB- and FMRFamide-like immunoreactivities in lobster neurons: Colocalization of distinct peptides or colabeling of the same peptide(s)?

Virtually all of the SCPB-like immunoreactive neurons (ca. 60 cells) in the lobster Homarus americanus also contain FMRFamide-like immunoreactivity. Control experiments reveal that SCPB-and FMRFamide-like immunoreactivities are successfully preadsorbed with their specific antigens, while the normal staining pattern is retained following preadsorption of each antibody with the alternate peptide. These experiments potentially lead to the conclusion that the anti-SCPB and anti-FMRFamide antibodies are labeling distinct compounds that are colocalized in lobster neurons. The lobster nervous system does not, however, contain authentic FMRFamide, but rather several FMRFamide-like compounds (Trimmer et al., J. Comp. Neurol. 266:16-26, 1987). The most abundant of these is the octapeptide TNRNFLRFamide. Experiments demonstrate that SCPB-like immunoreactivity is completely preadsorbed with synthetic TNRNFLRFamide, while there is a significant or complete loss of staining after preadsorption of the FMRFamide antibody with this molecule. Met-enkephalin-Arg-Phe-amide (YGGFMRFamide), an extended opioid peptide containing the FMRFamide sequence, also preadsorbs SCPB- and FMRFamide-like immunoreactivities, while Met-enkephalin-Arg-Phe (YGGFMRF) has no effect on the staining properties of these antibodies. These results suggest that the SCPB antibody can bind to extended forms of FMRFamide-like molecules, and that anti-SCPB and anti-FMRFamide antibodies may be colabeling one or more FMRFamide-like molecules in lobster neurons.     

Differential expression and function of synaptotagmin 1 isoforms in Caenorhabditis elegans

Synaptotagmin 1, encoded by the snt-1 gene in Caenorhabditis elegans, is a major synaptic vesicle protein containing two Ca(2+)-binding (C2) domains. Alternative splicing gives rise to two synaptotagmin 1 isoforms, designated SNT-1A and SNT-1B, which differ in amino acid sequence in the third, fourth, and fifth beta-strands of the second C2 domain (C2B). We report here that expression of either SNT-1 isoform under control of a strong pan-neural promoter fully rescues the snt-1 null phenotype. Furthermore, C-terminal fusions of either isoform with GFP are trafficked properly to synapses and are fully functional, unlike synaptotagmin 1Colon, two colonsGFP fusions in mice. Analysis of isoform expression with genomic GFP reporter constructs revealed that the SNT-1A and-1B isoforms are differentially expressed and localized in the C. elegans nervous system. We also report molecular, behavioral, and immunocytochemical analyses of twenty snt-1 mutations. One of these mutations, md259, specifically disrupts expression of the SNT-1A isoform and has defects in a subset of synaptotagmin 1-mediated behaviors. A second mutation, md220, is an in-frame 9-bp deletion that removes a conserved tri-peptide sequence (VIL) in the second beta-strand of the C2B domain and disrupts the proper intracellular trafficking of synaptotagmin. Site-directed mutagenesis of a functional SNT-1Colon, two colonsGFP fusion protein was used to examine the potential role of the VIL sequence in synaptotagmin trafficking. Although our results suggest the VIL sequence is most likely not a specific targeting motif, the use of SNT-1Colon, two colonsGFP fusions has great potential for investigating synaptotagmin trafficking and localization.     

Identification and characterization of Caenorhabditis elegans palmitoyl protein thioesterase1

Infantile neuronal ceroid lipofuscinosis (INCL; Batten disease) is a severe neurodegenerative disorder of childhood characterized by the accumulation of autofluorescent storage material in lysosomes. It is caused by mutation of the CLN1/PPT1 gene, which encodes the lysosomal enzyme palmitoyl protein thioesterase-1 (PPT1), but the mechanism of disease pathogenesis and substrates for the enzyme are unknown. Caenorhabditis elegans is a simple nematode worm, with a fully sequenced genome, which is easy to maintain and manipulate. It has a completely mapped cell lineage and nervous system and has already provided clues about the pathogenesis of several human neuronal and lysosomal storage disorders. We have identified and characterized a PPT1 homologue in C. elegans. We found that, although this gene was not essential for the animal's survival, its mutation resulted in a mild developmental and reproductive phenotype, affected the number and size of mitochondria, and resulted in an abnormality in mitochondrial morphology, possibly suggestive of a role for this organelle in INCL pathogenesis. This strain, deleted for ppt-1, potentially provides a model system for the study of PPT1 and the pathogenesis of INCL.     

Mutations in the Caenorhabditis elegans dystrophin-like gene dys-1 lead to hyperactivity and suggest a link with cholinergic transmission

Mutations in the human dystrophin gene cause Duchenne muscular dystrophy, a common neuromuscular disease leading to a progressive necrosis of muscle cells. The etiology of this necrosis has not been clearly established, and the cellular function of the dystrophin protein is still unknown. We report here the identification of a dystrophin-like gene (named dys-1) in the nematode Caenorhabditis elegans. Loss-of-function mutations of the dys-1 gene make animals hyperactive and slightly hypercontracted. Surprisingly, the dys-1 mutants have apparently normal muscle cells. Based on reporter gene analysis and heterologous promoter expression, the site of action of the dys-1 gene seems to be in muscles. A chimeric transgene in which the C-terminal end of the protein has been replaced by the human dystrophin sequence is able to partly suppress the phenotype of the dys-1 mutants, showing that both proteins share some functional similarity. Finally, the dys-1 mutants are hypersensitive to acetylcholine and to the acetylcholinesterase inhibitor aldicarb, suggesting that dys-1 mutations affect cholinergic transmission. This study provides the first functional link between the dystrophin family of proteins and cholinergic transmission. Received: August 18, 1998 / Accepted: September 15, 1998 / Published online: December 9, 1998     

The connectome of the Caenorhabditis elegans pharynx

Detailed anatomical maps of individual organs and entire animals have served as invaluable entry points for ensuing dissection of their evolution, development, and function. The pharynx of the nematode Caenorhabditis elegans is a simple neuromuscular organ with a self-contained, autonomously acting nervous system, composed of 20 neurons that fall into 14 anatomically distinct types. Using serial electron micrograph (EM) reconstruction, we re-evaluate here the connectome of the pharyngeal nervous system, providing a novel and more detailed view of its structure and predicted function. Contrasting the previous classification of pharyngeal neurons into distinct inter- and motor neuron classes, we provide evidence that most pharyngeal neurons are also likely sensory neurons and most, if not all, pharyngeal neurons also classify as motor neurons. Together with the extensive cross-connectivity among pharyngeal neurons, which is more widespread than previously realized, the sensory-motor characteristics of most neurons define a shallow network architecture of the pharyngeal connectome. Network analysis reveals that the patterns of neuronal connections are organized into putative computational modules that reflect the known functional domains of the pharynx. Compared with the somatic nervous system, pharyngeal neurons both physically associate with a larger fraction of their neighbors and create synapses with a greater proportion of their neighbors. We speculate that the overall architecture of the pharyngeal nervous system may be reminiscent of the architecture of ancestral, primitive nervous systems.     

Effect of dys‐1 mutation on gene expression profile in space‐flown Caenorhabditis elegans

Introduction: Dystrophin‐like dys‐1 gene expression increases in the body wall muscles of Caenorhabditis elegans after spaceflight (SF). Here we used a dys‐1(cx18) mutant to analyze the molecular adaptive responses of C. elegans to SF. Methods: DNA microarrays were performed to identify differentially expressed genes between wild‐type (WT) and dys‐1 mutant worms after SF. We performed Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses, predicted human diseases, and screened out key genes for human muscle diseases with NextBio. Results: Gene expression was less affected by SF in the dys‐1 mutant than in the WT worms. The dys‐1 mutation influenced neuromuscular gene expression (neuropeptide genes, muscle‐related genes, and dystrophin‐related genes) under SF conditions, among which 15 genes were specifically regulated by dys‐1. NextBio analysis predicted that cdka‐1, lev‐11, unc‐27, and unc‐94 genes might play critical roles in muscle atrophy. Discussion: dys‐1 Potentially regulates the neuromuscular system in space. Muscle Nerve 58 : 114–122, 2018     

Quantitative analysis of thermotaxis in the nematode Caenorhabditis elegans

Thermotaxis (TTX) is one of the sophisticated behaviors in the nematode Caenorhabditis elegans. Although the mechanisms of thermotaxis have been deduced from different studies, they are controversial. Previous studies proposed a behavioral model where thermotaxis is regulated by the counterbalance between two opposite driving forces, while recent studies proposed stochastic models. In this study, we analyzed thermotaxis by a novel quantitative population TTX assay using a gentle linear thermal gradient. Analysis of thermotaxis in wild type animals revealed a clear thermal preference to a cultivation temperature with regard to the distribution of animals and the TTX mean expressing temperature preference. A time course assay revealed that the behavioral response to a preferred temperature was initially suppressed for at least 15 min in the animals cultivated at 23 degrees C, but not in those cultivated at 17 degrees C. Our result provides a possible explanation for the inconsistency between the various studies on thermotaxis and is consistent with the early behavioral model, where thermotaxis is regulated by the counterbalance between two driving forces.     

模式动物线虫的学习与学习抉择行为

线虫因为其简单的体系和对于诸如接触、味道、气味和温度等外界刺激的敏感反映能力,日益成为受人瞩目的研究行为可塑性的模式动物。目前认为,线虫中的学习包括非联想与联想学习两大类,且存在至少6种形式化学趋向性介导的联想学习。同时,三种研究体系已经被开发出来用于学习抉择的研究。本文讨论了线虫中学习与学习抉择的形式、研究模型及其遗传与分子调控机制。     

Cholinergic receptor mutants of the nematode Caenorhabditis elegans

Potential acetylcholine receptor (AChR) mutants of the nematode are selectable by resistance to the neurotoxic drug levamisole, a probable cholinergic agonist. To determine which mutants may have achieved resistance through loss of levamisole receptor function, we have assayed mutant extracts for specific 3H-meta-aminolevamisole binding activity in the presence and absence of mecamylamine. We find that mutants in 3 of the 7 genes associated with extreme levamisole resistance are obviously deficient in saturable specific 3H-meta-aminolevamisole binding activity. Mutants of the 4 other genes have abnormal binding activities that fail to undergo the apparent allosteric activation of saturable specific 3H-meta-aminolevamisole binding activity caused by mecamylamine. Thus, all 7 genes appear to be required to produce a fully functional levamisole receptor. Mutants of several other genes associated only with partial resistance to levamisole have at least grossly normal receptor binding activities.     

模式动物线虫的学习与学习抉择行为(英文)

线虫因为其简单的体系和对于诸如接触、味道、气味和温度等外界刺激的敏感反映能力,日益成为受人瞩目的研究行为可塑性的模式动物。目前认为,线虫中的学习包括非联想与联想学习两大类,且存在至少6种形式化学趋向性介导的联想学习。同时,三种研究体系已经被开发出来用于学习抉择的研究。本文讨论了线虫中学习与学习抉择的形式、研究模型及其遗传与分子调控机制。     

秀丽线虫记忆的分子调控机制

模式无脊椎动物秀丽线虫已经成为揭示记忆复杂行为的理想研究模型之一.线虫具有三种简单的记忆形式对温度感知的记忆、对化学物质感知的记忆以及对于机械刺激感知的记忆.在对机械刺激感知的记忆研究中,短时程、中时程与长时程记忆均得到了系统的分析.其中短时程与中时程记忆可能定位于感觉神经元的前突触,而长时程记忆可能定位于中间神经元的后突触.本文针对线虫中记忆的遗传与分子调控机制近些年的研究进展进行了总结与讨论.     

Molecular control of memory in nematode Caenorhabditis elegans

Model invertebrate organism Caenorhabditis elegans has become an ideal model to unravel the complex processes of memory. C. elegans has three simple forms of memory： memory for thermosensation, memory for chemosensation, and memory for mechanosensation. In the form of memory for mechanosensation, short-term memory, intermediate-term memory, and long-term memory have been extensively studied. The short-term memory and intermediate-term memory may occur in the presynaptic sensory neurons, whereas the long-term memory may occur in the postsynaptic interneurons. This review will discuss the recent progress on genetic and molecular regulation of memory in C. elegans.