Wan Omar A et al. (2001) Malays J Med Sci "Epidemiological screening of lymphatic filariasis among immigrants using dipstick colloidal dye immunoassay."

Sulaiman O, Wan Omar A, Yusof S, Ismail G, Fatmah MS, Rahmah N, Khairul AA

[Malays J Med Sci, 2001]

We have recently reported that a dipstick colloidal dye immunoassay (DIA) that detect parasite antigens in human serum is sensitive and specific for the diagnosis of active infection of lymphatic filariasis. Rabbit polyclonal antibodies (RbBmCAg) labelled with a commercial dye, palanil navy blue was used to detect filarial antigenemia among Indonesian and Bangladeshi immigrant workers (N= 630) at oil palm estates at Hulu Trengganu District, Peninsular Malaysia. Microfilaremia with Brugia malayi were detected in 51 (8.10 %) individuals, of which 42 (6.67 %) were among the Indonesians and 9 (1.98 %) among the Bangladeshis. Microfilaremia with Wuchereria bancrofti were detected in 33 (5.24 %) individuals of which 15 (2.38 %) were among the Indonesians and 18 (2.86 %) among the Bangladeshis workers. The DIA detected 96 (15.24 %) antigenemic cases which comprise of all the microfilaremic cases and 15 (2.38 %) amicrofilaremic cases. The amicrofilaremic cases with filarial antigenemia consisted of 9 (1. 43 %) Indonesians and 6 (0.95%) Bangladeshis. We have used 6 ul of the RbBmCAg and diluted (1:10) patients' sera per dipstick which make the DIA reagent conservative. The DIA is a rapid test and can be read in approximate 2 hours.. Additionally, coloured dots developed in the DIA can be qualitatively assessed visually for intensity. The DIA does not require sophisticated equipment or radioactivity, and therefore suitable for field application.

Shen Q et al. (2009) Mol Cell Biol "Adenine nucleotide translocator cooperates with core cell death machinery to promote apoptosis in Caenorhabditis elegans."

Yang C, Xu Z, Xiao H, Gao Z, Qin F, Shen Q, Cui J

[Mol Cell Biol, 2009]

In Caenorhabditis elegans, the central cell-killing process is essentially controlled by the interplay of four apoptotic factors: EGL-1/BH3-only protein, CED-9/Bcl2, CED-4/Apaf1, and CED-3/caspase. In cells destined to die, EGL-1 binds to CED-9 and results in the release of CED-4 from the mitochondrion-tethered CED-9-CED-4 complex to the perinucleus, which facilitates processing of the CED-3 caspase to cause apoptosis. However, whether additional factors exist to regulate the cell-killing process remains largely unknown. We have identified here WAN-1, the C. elegans ortholog of mammalian adenine nucleotide translocator, as an important cell death regulator. Genetic inactivation of wan-1 significantly suppressed both somatic and germ line cell deaths in C. elegans. Consistently, chemical inhibition of WAN-1 activity also caused strong reduction of germ line apoptosis. WAN-1 localizes to mitochondria and can form complex with both CED-4 and CED-9. Importantly, the cell death initiator EGL-1 can disrupt the interaction between CED-9 and WAN-1. In addition, overexpression of WAN-1 induced ectopic cell killing dependently on the core cell death pathway. These findings suggest that WAN-1 is involved in the central cell-killing process and cooperates with the core cell death machinery to promote programmed cell death in C. elegans.

Qinfang Shen et al. (2008) "Adenine nucleotide translocator cooperates with core cell death machinery to control programmed cell death in C. elegans"

Zhiheng Xu, Qinfang Shen, Zhiyang Gao, Fengsong Qin, Chonglin Yang, Jie Cui

[2008]

"In C. elegans, the central cell-killing process is essentially controlled by the interplay of four apoptotic factors: EGL-1/BH3-only protein, CED-9/Bcl-2, CED-4/Apaf-1 and CED-3/caspase. In cells that are destined to die, the BH3-only proapoptotic protein EGL-1 is up regulated and binds to CED-9, resulting in the release of CED-4 from the mitochondria-tethered CED-9-CED-4 complex to perinucleus, which promotes the processing of CED-3 and leads to apoptosis. However, whether additional regulators exist to regulate the cell killing process together with these 4 apoptotic factors remains largely unknown. Here we have identified WAN-1, the C. elegans ortholog of mammalian adenine nucleotide translocator 1 (ANT1), as an important cell death regulator in C. elegans. WAN-1 localizes to mitochondria and forms complex with either CED-4 or CED-9 but not EGL-1. Importantly, the cell death initiator EGL-1 can disrupt the CED-9-WAN-1 complex. In activation of wan-1 by RNA interference significantly suppresses both somatic and germline apoptosis. Consistently, inhibition of WAN-1/ANT activity by Bongkrekic acid, an ANT-specific inhibitor, also leads to drastic suppression of germline apoptosis. These findings suggest that WAN-1/ANT is directly involved in the cell-killing process and cooperates with core cell death components to control the activation of apoptosis. Moreover, our results suggest that adenine nucleotide translocator functions evolutionarily to regulate cell death from C. elegans to humans."

Chan S et al. (1994) Worm Breeder's Gazette "The unc-40 Axon Guidance Gene Encodes a Transmembrane Receptor With Extracellular Fibronectin and Immunoglobulin Domains"

Culotti J, Chan S, Zheng H, Su M-W

[Worm Breeder's Gazette, 1994]

The unc-40 axon guidance gene encodes a transmembrane receptor with extracellular fibronectin and immunoglobulin domains. Shirley Chan, Ming-Wan Su, Hong Zhang, Joe Culotti Samuel Lunenfeld Research Institute of Mt. Sinai Hospital Toronto, Canada Ed Hedgecock, Johns Hopkins University, Baltimore unc40 primarily affects ventrally oriented pioneer growth cone and cell migrations that occur on the epideImis. unc40 also has rninor effects on dorsally oriented pioneer migrations (Hedgecock et al., Neuron 4:61-85,1990). We mapped unc40 relative to strain-specific DNA polymorphisms, then rescued the Unc and DTC defects by germline transformation with a cosmid and with a 14 kb phage from the region. Based on germline transformation experiments using overlapping cosmid and phage clones, we were able to identify a region required for rescue that encodes a single transcript by Northern analysis of 5.5 kb. We cloned and sequenced a 4.2 kb cDNA (S. Kim library) corresponding to this transcript. This cDNA contains a 3.3kb open reading frame. The predicted unc- 40 protein (so far) has a single hydrophobic transmembrane domain, six fibronectin type III (FNIII) domains and two immunoglobulin (IG) domains. FNIII and IG domains are found in a variety of transmembrane proteins, but these most commonly have three or five FNIII domains. Several transmembrane proteins that have extracellular IG and FNIII domains also have intracellular tyrosine kinase or phosphotyrosine phophatase domains, but the predicted 309 residue intracellular region of unc-40 has no obvious similarity to these domains. We are currently sequencing genomic DNA to identify the 5' end of unc40. Northern and Southern analyses have failed to reveal detectable deletions or rearrangements in a dozen unc40 mutants analyzed. In addition to affecting migrations on the dorsoventral coordinates of the epidermis, unc-40 mutations also affect body shape, normally an epidermal function. To determine where unc-40 is expressed and where it is required for guidance, we are constructing strains to do mosaic analysis and to analyze patterns of expression from the unc-40 promoter.

Chan S et al. (1995) Worm Breeder's Gazette "unc-40 Encodes a Protein Related to the Human DCC (Deleted Colorectal Cancer) Protein."

Su M-W, Culotti JG, Hedgecock EM, Chan S

[Worm Breeder's Gazette, 1995]

unc-40 ENCODES A PROTEIN RELATED TO THE HUMAN DCC (DELETED COLORECTAL CANCER) PROTEIN. Shirley Chan, Ming-Wan Su, and Joe Culotti Samuel Lunenfeld Research Institute of Mt. Sinai Hospital Toronto, Canada Ed Hedgecock Department of Biology Johns Hopkins University Baltimore, MD unc-40 is required for circumferential pioneer migrations of axon growth cones and mesodermal cells on the basal surface of the epidermis of C. elegans. Although unc-40 mutations primarily affect migrations oriented towards the ventral side, they also affect dorsally-oriented migrations (although to a lesser extent). The unc-40 gene was cloned by mapping unc-40 mutations relative to DNA polymorphisms followed by transformation rescue. We have determined what we believe may be the entire coding region of this gene comprising 4 kb of sequence. From amino to carboxy terminus, the predicted UNC40 protein has 4 immunoglobulin domains, 6 fibronectin type III domains, a single transmembrane domain and an intracellular domain of 309 amino acids. The overall domain structure of the predicted UNC-40 protein is very similar to the human DCC (deleted colorectal cancer) protein which was initially (incorrectly?) implicated in a fraction of human colorectal cancers. More recently, a DCC-related gene has been cloned from chicken (Jost Vielmetter and William Dryer, Cal Tech, personal communication) called neogenin. neogenin is not the chicken DCC gene, but is related to it. The fibronectin and immunoglobulin domains of UNC-40 are clearly more related to DCC and Neogenin than they are to similar domains in other proteins. Interestingly, the Neogenin protein is expressed in neurons as they extend axons in the developing chick nervous system. We are currently attempting several methods for examining the expression pattern of the UNC-40 protein.

Ouyang, John Paul et al. (2021) International Worm Meeting "Independent nuclear and cytoplasmic silencing mechanisms contribute to transgenerational RNAi"

Ouyang, John Paul, Seydoux, Geraldine, Zhang, Lucy

[International Worm Meeting, 2021]

RNA-mediated interference is a conserved mechanism used broadly to regulate gene expression. In C. elegans, RNAi is heritable and has been shown to target both nascent transcripts in the nucleus and mature transcripts in the cytoplasm (Billi et al., 2014). To explore how targeting of nuclear and cytoplasmic RNAs is coordinated during RNAi, we used fluorescence in situ hybridization (FISH) to examine the distribution of germline mRNAs targeted by feeding-induced RNAi. We observed a transient increase in nascent transcripts 4-8 hours after the onset of feeding followed by a subsequent decrease. Cytoplasmic transcripts accumulated in nuage (perinuclear condensates) and were rapidly turned over in the bulk cytoplasm. In progeny of fed mothers, we observed reduced transcript levels at the locus, low levels of RNA present in the nuage, and RNA strongly depleted in the bulk cytoplasm. These findings confirm that RNAi affects the accumulation and distribution of nascent and cytoplasmic transcripts in both animals exposed to the RNAi trigger and their progeny. HRDE-1/WAGO-9 is a nuclear Argonaute required for transgenerational RNAi (Buckley et al., 2012). As expected, we found that HRDE-1 is required for nascent transcripts to respond to RNAi. Despite no change in nascent transcripts, hrde-1 mutants accumulated targeted mRNAs in the nuage in both fed mothers and F1 progeny as in wild-type. ZNFX-1 is a nuage-localized SF1 helicase also required for transgenerational inheritance of RNAi (Ishidate et al., 2018; Wan et al., 2018). We found that znfx-1 mutants have a phenotype opposite that of hrde-1: a wild-type nuclear response, but a failure in both recruiting RNAs to nuage and degrading the RNAs in the cytoplasm. Consistent with HRDE-1 and ZNFX-1 functioning in distinct branches of the RNAi pathway, znfx-1; hrde-1 double mutants were defective in both nuclear and cytoplasmic responses and failed to show any silencing in F1 progeny. These observations suggest that RNAi triggers two distinct responses required for transgenerational RNAi: a nuclear response dependent on hrde-1 that reduces production/accumulation of nascent RNAs at the targeted locus, and a cytoplasmic response dependent on znfx-1 that retains targeted mRNAs in nuage and prevents their accumulation in the cytoplasm. The nuclear and cytoplasmic responses can be inherited independently of one another, and both are required for maximum silencing in progeny.

Uebel, Celja J. et al. (2021) MicroPubl Biol

Manage, Kevin I., Uebel, Celja J., Phillips, Carolyn M.

[MicroPubl Biol, 2021]

Multiple condensates occupy the perinuclear space of C. elegans germ cells, where they coordinate RNA surveillance to ensure proper gene expression (Lev and Rechavi 2020; Sundby et al. 2021). The most well-studied of these condensates are P granules, phase-separated germ granules required for maintenance of germ cell identity and fertility (Kawasaki et al. 1998; Updike et al. 2014). P granule morphology and localization is well documented in C. elegans development (Strome et al. 1982). Adjacent to P granules are Mutator foci, which are nucleated by MUT-16 and required for the amplification of small interfering RNAs (siRNAs) to create a robust and heritable silencing signal (Phillips et al. 2012). During development, faint Mutator foci are occasionally seen in the P4 germline blastomere of 30-cell embryos, but are most robust and numerous in the Z2 and Z3 progenitor germ cells (PGCs) of 100-cell embryos (Uebel et al. 2020). A third germline condensate, Z granules, are situated between P granules and Mutator foci and facilitate transgenerational epigenetic inheritance of silencing signals. Z granule components ZNFX-1 and WAGO-4 colocalize with P granules in early embryos, but begin to de-mix from P granules in the Z2/Z3 PGCs to form separate Z granule condensates (Wan et al. 2018). Lastly, recently discovered SIMR foci also intimately localize within this cluster of germline granules. SIMR-1, a key component of SIMR foci, is a Tudor domain protein that mediates production of secondary siRNAs for piwi-interacting RNA (piRNA)-targeted mRNAs (Manage et al. 2020). While P granule, Z granule, and Mutator foci localization through embryonic development has been previously described, the embryonic appearance of SIMR foci is not known.

Quartey MO et al. (2021) Sci Rep "The A(1-38) peptide is a negative regulator of the A(1-42) peptide implicated in Alzheimer disease progression."

Pennington PR, Heistad RM, Nyarko JNK, Barnes JR, Bolanos MAC, Parsons MP, Knudsen KJ, De Carvalho CE, Leary SC, Mousseau DD, Buttigieg J, Maley JM, Quartey MO

[Sci Rep, 2021]

The pool of -Amyloid (A) length variants detected in preclinical and clinical Alzheimer disease (AD) samples suggests a diversity of roles for A peptides. We examined how a naturally occurring variant, e.g. A(1-38), interacts with the AD-related variant, A(1-42), and the predominant physiological variant, A(1-40). Atomic force microscopy, Thioflavin T fluorescence, circular dichroism, dynamic light scattering, and surface plasmon resonance reveal that A(1-38) interacts differently with A(1-40) and A(1-42) and, in general, A(1-38) interferes with the conversion of A(1-42) to a -sheet-rich aggregate. Functionally, A(1-38) reverses the negative impact of A(1-42) on long-term potentiation in acute hippocampal slices and on membrane conductance in primary neurons, and mitigates an A(1-42) phenotype in Caenorhabditis elegans. A(1-38) also reverses any loss of MTT conversion induced by A(1-40) and A(1-42) in HT-22 hippocampal neurons and APOE 4-positive human fibroblasts, although the combination of A(1-38) and A(1-42) inhibits MTT conversion in APOE 4-negative fibroblasts. A greater ratio of soluble A(1-42)/A(1-38) [and A(1-42)/A(1-40)] in autopsied brain extracts correlates with an earlier age-at-death in males (but not females) with a diagnosis of AD. These results suggest that A(1-38) is capable of physically counteracting, potentially in a sex-dependent manner, the neuropathological effects of the AD-relevant A(1-42).

Danielle Thierry-Mieg et al. (2003) Worm Breeder's Gazette "www.wormgenes.org , a new gene centered database"

Vahan Simonyan, Michel Potdevin, Mark Sienkiewicz, Yuji KOHARA, Jean Thierry-Mieg, Danielle Thierry-Mieg, Tadasu SHIN-I

[Worm Breeder's Gazette, 2003]

Wormgenes is a new resource for C.elegans offering a detailed summary about each gene and a powerful query system.

Joseph D Watson et al. (2005) International Worm Meeting "The synaptic protein, RPM-1, regulates neuronal gene expression."

Stephen E Von Stetina, Joseph D Watson, David M Miller

[International Worm Meeting, 2005]

With its well-defined neuronal architecture and powerful genetics, C. elegans is a useful model system for studies of neuronal differentiation. In an effort to identify genes that specify motor neuron fate, we screened for mutations that selectively reduce expression of acr-5::YFP in B-class motor neurons. Surprisingly, three of these isolates (wd55, wd67, wd72) are new alleles of rpm-1 (Regulator of Presynaptic Morphology). The RPM-1 protein, and its homologues Highwire (Hiw) (Drosophila) and PHR1 (vertebrates) are localized to presynaptic regions adjacent to the active zone where they are believed to function as E3 ubiquitin ligases (Zhen, Jin et al 2004) (Wan, Goodman et al 2000). Mutations in rpm-1/hiw/Phr1 disrupt synaptic structure and function. This effect may result from the loss of ubiquitin-dependent protein degradation (Liao, Hung et al. 2004) (McCabe, Hom et al. 2004). Genetic epistasis experiments indicate that RPM-1 normally downregulates pmk-3 (p38 MAPK) signaling (K. Nakata and Y. Jin, et al. 2005). We have now shown that pmk-3 also suppresses disregulation of B-class motor neuron markers in rpm-1 mutants. This finding suggests that RPM-1, a component of the presynaptic apparatus, regulates transcription of specific target genes via a MAPK signaling pathway. We have used the mRNA tagging method (Roy and Kim, et al. 2002) to identify downstream targets of rpm-1. A pan neural mRNA tagging line (F25B3.3::FLAG::PAB-1) was used to isolate mRNA from all neurons in the rpm-1 deletion mutant, ok364; ~100 genes are enriched (> 2x) and ~50 genes are depleted (> 2x) relative to wildtype in rpm-1 mutant neurons. We have confirmed these effects for selected genes by quantitative RT-PCR. We are currently using RNAi in a sensitized genetic background to test candidate genes from these lists for roles in RPM-1 function. This work is expected to reveal downstream targets of RPM-1 with key roles in synaptogenesis.