Sangvaranond A et al. (1989) Zentralbl Veterinarmed B "19s and 7s antibody response of Mastomys natalensis in experimental filarial (Litomosoides carinii, Acanthocheilonema viteae, Brugia malayi, B. pahangi) infections."

Sangvaranond A, Zahner H

[Zentralbl Veterinarmed B, 1989]

Specific total antibody (ab), 19s and 7s ab levels in the serum of M. natalensis were investigated after infection with L. carinii, A. viteae, B. malayi and B. pahangi for period of about 500 days p.i., using ELISA (homologous adult antigen) and indirect immunofluorescence tests (IIFT: homologous adult and microfilariae antigen). Total ab levels in L. carinii infected animals rose moderately during prepatency Maximum levels occurred during patency. The response during prepatency was stronger in A. viteae and Brugia spp. infected hosts. Lateron ab levels increased continuously in Brugia infections; in A. viteae infection they decreased with decreasing parasitaemia. 19s abs were stimulated during prepatency and at the beginning of patency, or were found at moderate levels throughout to period of investigation (Brugia infections). 7s abs predominated beginning at the period of late prepatency (IIFT) or at the beginning of patency (ELISA). The time courses of 7s abs corresponded to those of total abs. As obvious by IIFT (adult worm antigen) total and 19s titres were higher against cuticle antigens, egg shell antigens and intrauterine amorphous material than against antigens located in the hypodermis and musculature. 7s abs showed best reactivity with cuticle antigens. Using microfilarial antigens 19s abs reacted predominantly with cuticle antigens whereas 7s abs often showed higher titres against antigens which were localized within the larvae than against cuticle antigens.

Hu M et al. (2007) Exp Parasitol "Trichostrongylus vitrinus (Nematoda: Strongylida): molecular characterization and transcriptional analysis of Tv-stp-1, a serine/threonine phosphatase gene."

Gasser RB, Abs El-Osta YG, Campbell BE, Beveridge I, Hu M, Boag PR, Nisbet AJ

[Exp Parasitol, 2007]

A full-length cDNA (Tv-stp-1) encoding a serine/threonine protein phosphatase (Tv-STP-1) was isolated from Trichostrongylus vitrinus (order Strongylida), an economically important parasitic nematode of small ruminants. The uninterrupted open reading frame (ORF) of 951 nucleotides encoded a predicted protein of 316 amino acids (aa), containing the characteristic motif [LIVMN]-[KR]-G-N-H-E. Comparison with other sequences in non-redundant databases showed that Tv-STP-1 had significant identities/similarities to those from a range of metazoans and protists. Sequence similarity was most pronounced in the central region of the protein, in which the catalytic activity is inferred to be modulated by eight conserved residues (Asp 61, His 63, Asp 92, Asp 95, Asn 121, His 171, His 246 and Tyr 270), known to coordinate the binding of two metal ions (Mn(2+) and Fe(2+)) in various organisms. Phylogenetic analyses of selected amino acid sequence data using the neighbor-joining and maximum parsimony methods revealed Tv-STP-1 to be most closely related to the glc seven-like phosphatases inferred for genes from the free-living nematode Caenorhabditis elegans and the parasitic nematode Oesophagostomum dentatum (order Strongylida). Comparison of the genomic organization of the full-length Tv-stp-1 gene with related molecules from other nematodes revealed substantial variation in the lengths and numbers of the exons and introns. The entire genes Tv-stp-1 (5041-5362bp; 10 exons and 9 introns) and Od-mpp-1 (10,271bp; 8 exons and 9 introns) from the parasitic nematodes T. vitrinus and O. dentatum were considerably longer than the C. elegans genes (1222-1603 bp; 3-7 exons and 2-6 introns). Transcriptional analysis by reverse transcription polymerase chain reaction (RT-PCR) showed that Tv-stp-1 was transcribed in adult males of T. vitrinus, but not in the adult female or in any larval stages of this species. In spite of considerable variation at the genomic level, the findings of the present study suggest that there is relative conservation in features and function of the serine/threonine protein phosphatase characterized among T. vitrinus, O. dentatum and C. elegans, which should have implications for exploring molecular reproductive and developmental processes in strongylid nematodes of socio-economic importance.

El Mouridi, Sonia et al. (2021) MicroPubl Biol

AlHarbi, Sarah, El Mouridi, Sonia, Frkjr-Jensen, Christian

[MicroPubl Biol, 2021]

For El Mouridi, S; AlHarbi, S; Frkjr-Jensen, C (2021). A histamine-gated channel is an efficient negative selection marker for C. elegans transgenesis. microPublication Biology. 10.17912/micropub.biology.000349.

Olivier-Mason A et al. (2013) Development "Transmembrane protein OSTA-1 shapes sensory cilia morphology via regulation of intracellular membrane trafficking in C. elegans."

Blacque OE, Nechipurenko IV, Bowie RV, Wojtyniak M, Olivier-Mason A, Sengupta P

[Development, 2013]

The structure and function of primary cilia are critically dependent on intracellular trafficking pathways that transport ciliary membrane and protein components. The mechanisms by which these trafficking pathways are regulated are not fully characterized. Here we identify the transmembrane protein OSTA-1 as a new regulator of the trafficking pathways that shape the morphology and protein composition of sensory cilia in C. elegans. osta-1 encodes an organic solute transporter alpha-like protein, mammalian homologs of which have been implicated in membrane trafficking and solute transport, although a role in regulating cilia structure has not previously been demonstrated. We show that mutations in osta-1 result in altered ciliary membrane volume, branch length and complexity, as well as defects in localization of a subset of ciliary transmembrane proteins in different sensory cilia types. OSTA-1 is associated with transport vesicles, localizes to a ciliary compartment shown to house trafficking proteins, and regulates both retrograde and anterograde flux of the endosome-associated RAB-5 small GTPase. Genetic epistasis experiments with sensory signaling, exocytic and endocytic proteins further implicate OSTA-1 as a crucial regulator of ciliary architecture via regulation of cilia-destined trafficking. Our findings suggest that regulation of transport pathways in a cell type-specific manner contributes to diversity in sensory cilia structure and might allow dynamic remodeling of ciliary architecture via multiple inputs.

Po MD et al. (2012) Neuron "Releasing the inner inhibition for axon regeneration."

Po MD, Calarco JA, Zhen M

[Neuron, 2012]

The adult mammalian central nervous system exhibits restricted regenerative potential. Chen etal. (2011) and El Bejjani and Hammarlund (2012) used Caenorhabditis elegans to uncover intrinsic factors that inhibit regeneration of axotomized mature neurons, opening avenues for potential therapeutics.

Todd Heallen et al. (2006) Development & Evolution Meeting "Novel Regulation of the Aurora B Kinase Pathway by a Conserved Member of the AAA-ATPase Protein Family"

Todd Heallen, Jill Schumacher

[Development & Evolution Meeting, 2006]

Timely and error-free progression of the cell cycle is fundamental to the development and proper function of eukaryotic organisms. As such, eukaryotes have evolved complex regulatory mechanisms to facilitate development and cell maintenance. Our studies are focused on an evolutionarily conserved family of mitotic proteins called the Aurora kinases. These proteins comprise a class of serine/threonine kinases that are essential for mitotic events such as centrosome maturation, spindle assembly, chromosome segregation, and cytokinesis (Andrews et. al., 2003). AIR-2, the C. elegans homolog of Aurora B, has been shown to interact with a variety of substrates to facilitate processes such as chromosome segregation and cytokinesis. Depletion of AIR-2 via RNA-mediated interference (RNAi) leads to embryonic lethality at the one cell stage (Schumacher et. al., 1998). A temperature-sensitive (ts) mutation, air-2 (or207ts) confers a similar phenotype at restrictive temperatures (Severson et. al., 2000). Our current goal is to elucidate the AIR-2 regulatory pathway by identifying the components working in concert with AIR-2. We therefore conducted a genome-wide RNAi screen to identify suppressors of air-2(or207ts) lethality. We have screened through an RNAi construct library comprising nearly every gene in the C. elegans genome (approximately 17,000 genes). The strongest RNAi suppressor identified from this screen encodes a putative C. elegans AAA-ATPase and confers ~72.6% viability to the air-2(or207ts) strain at 20ºC as compared with 1% viability for air-2(or207ts) alone. For simplicity purposes, we have tentatively named this suppressor abs-1 (Aurora B Suppressor-1). To understand the relationship between air-2 and abs-1, functional analyses are being performed. In vitro binding and kinase assays reveal that ABS-1 can bind directly to AIR-2 and inhibit its kinase activity. In addition, Western analysis revealed that AIR-2 levels are stabilized in abs-1(RNAi) embryo extracts as compared to controls. Furthermore, immunostaining studies reveal that AIR-2 localization is disrupted in abs-1(RNAi) embryos. Hence, our data suggests that abs-1(RNAi) can suppress the air-2(or207ts) lethality by stabilizing AIR-2 levels and by controlling the localization and kinase activity of AIR-2. We are therefore examining the possibility that the AIR-2 complex is regulated by ABS-1 via proteolytic and non-proteolytic pathways.

Bourdon JC et al. (1997) Oncogene "Further characterisation of the p53 responsive element--identification of new candidate genes for trans-activation by p53."

Debuire B, Dessen P, Bourdon JC, Deguin-Chambon V, May E, May P, Lelong JC

[Oncogene, 1997]

The p53 protein is known to trans-activate a number of genes by specific binding to a consensus sequence containing two decamers of the type: PuPuPuCA/TT/AGPyPyPy. In order to identify new p53 trans-activated genes, we defined a set of criteria for computer search of p53-responsive elements. Based on experimental data, we proposed an extended consensus sequence composed of the two decamers of the El-Deiry consensus sequence flanked by two additional ones. A maximum of 3 bp substitutions was accepted for the two decamers of the El-Deiry consensus sequence, as well as for each additional decamer, except when the two decamers of the El-Deiry consensus sequence are contiguous. In this case, each additional decamer is allowed to bear one base insertion or deletion between the median C and G. This set of criteria was validated by identifying within the promoter region of the IGF-BP3 gene the existence of a novel p53-responsive element whose functional significance was verified. By limiting our computer search to Vertebrate genes involved in cell cycle regulation, cellular adhesion or metastatic processes and to gene families most often found in HOVERGEN database, 7785 gene sequences were first analysed. Among the oncogenes, kinases, proteases and structural proteins, 55 new genes were selected; six of them were retrieved in more than one species.

Tabuse Y et al. (1983) Worm Breeder's Gazette "more monoclonal antibodies to germline granules."

Miwa J, Tabuse Y

[Worm Breeder's Gazette, 1983]

In a previous newsletter (WBG 7[1]:75), we described the making of a library of monoclonal antibodies (MoAbs) to C. elegans in order to identify tissue- or cell-specific antigens in the animal. We recently reported on three germline-specific MoAbs, two of which react with granules unique to germline cells (Yamaguchi et al. 1983. Develop., Growth and Differ. 25:121-131). In an attempt to obtain more germline-specific (and also early embryo-specific) MoAbs, we used fertilized eggs and 1- to 4-cell stage embryos for immunization. Of 800 hybridomas obtained by fusing P3U-1 myeloma cells to splenocytes from immunized BALB/c mice, about 60 hybridomas were found to secrete antibodies to C. elegans by indirect immunofluorescent staining of C. elegans whole mount preparations. Five hybridoma lines, 704D, 904A, 911F, 1003B, and 1108E, out of these 60 were identified as secretors of Abs that react with germline granules (G-granules). We are at present characterizing these cell lines together with the line M32 isolated previously. The table shows preliminary results on the cross-reactivity of the Abs to various nematodes. The Abs of all 6 hybridomas reacted with the G-granules of both C. elegans and C. briggsae, possibly reflecting the taxonomic closeness. However, the Ab of only one hybridoma line, 1003B, stained G-granules of a free-living diecious nematode recently isolated from soil of Wakayama-ken. The nematode Bursaphelenchus is parasitic and considered a prime culprit of recent disastrous damage to pine trees in Japan. The G-granules of this nematode were stained also with the 1003B Ab and tentatively with 904A and M32. The result with the 1003B Ab indicates that the G-granules of all the nematodes listed here possess some structure common to them. The difference observed of cross-reactivity among the Abs may reflect either various antigenic determinants of a single element in the granules or a complex of multiple elements constituting the granules. Further study on cross-reactivity and biochemical characterization are under way. [See Figure 1]

Neline Kriek et al. (2002) European Worm Meeting "The mechanism of action of the neuropeptide AF1 in C.elegans"

David Marsden, Neline Kriek, Julian Burke, Candida Rogers, Lindy Holden-Dye

[European Worm Meeting, 2002]

AF1 (KNEFIRFamide) was the first neuropeptide to be isolated from a nematode 1. It has a potent stimulatory action on the body wall muscle of Ascaris. Furthermore, it has a potent stimulatory action on the pharyngeal muscle of C.elegans 2. A C.elegans gene, flp-8, encoding multiple copies of AF1 has been identified 3, however deletion of this gene does not result in any obvious phenotype (Kim & Li, Soc. Neurosci. Abs. 1999). To delineate the putative physiological role of this peptide in the control of locomotion and feeding, and to define the signalling pathway, we are comparing the response to AF1 in wild type and mutant strains.

Michael A Beer (2005) International Worm Meeting "Systematic genome-wide identification of transcriptional cis-regulatory logic in C. elegans."

Michael A Beer

[International Worm Meeting, 2005]

We have developed a systematic approach for inferring cis-regulatory logic from whole-genome microarray expression data.[1] This approach identifies local DNA sequence elements and the combinatorial and positional constraints that determine their context-dependent role in transcriptional regulation. We use a Bayesian probabilistic framework that relates general DNA sequence features to mRNA expression patterns. By breaking the expression data into training and test sets of genes, we are able to evaluate the predictive accuracy of our inferred Bayesian network. Applied to S. cerevisiae, our inferred combinatorial regulatory rules correctly predict expression patterns for most of the genes. Applied to microarray data from C. elegans[2], we identify novel regulatory elements and combinatorial rules that control the phased temporal expression of transcription factors, histones, and germline specific genes during embryonic and larval development. While many of the DNA elements we find in S. cerevisiae are known transcription factor binding sites, the vast majority of the DNA elements we find in C. elegans and the inferred regulatory rules are novel, and provide focused mechanistic hypotheses for experimental validation. Successful DNA element detection is a limiting factor in our ability to infer predictive combinatorial rules, and the larger regulatory regions in C. elegans make this more challenging than in yeast. Here we extend our previous algorithm to explicitly use conservation of regulatory regions in C. briggsae to focus the search for DNA elements. In addition, we expand the range of regulatory programs we identify by applying to more diverse microarray datasets.[3] 1. Beer MA and Tavazoie S. Cell 117, 185-198 (2004). 2. Baugh LR, Hill AA, Slonim DK, Brown EL, and Hunter, CP. Development 130, 889-900 (2003); Hill AA, Hunter CP, Tsung BT, Tucker-Kellogg G, and Brown EL. Science 290, 809812 (2000). 3. Baugh LR, Hill AA, Claggett JM, Hill-Harfe K, Wen JC, Slonim DK, Brown EL, and Hunter, CP. Development 132, 1843-1854 (2005); Murphy CT, McCarroll SA, Bargmann CI, Fraser A, Kamath RS, Ahringer J, Li H, and Kenyon C. Nature 424 277-283 (2003); Reinke V, Smith HE, Nance J, Wang J, Van Doren C, Begley R, Jones SJ, Davis EB, Scherer S, Ward S, and Kim SK. Mol Cell 6 605-616 (2000).