Matsumoto, Shinya et al. (2015) International Worm Meeting "Analysis of intergenerational effects of starvation."

Matsumura, Yasuki, Matsumoto, Shinya, Morishita, Mayu, Sei, Megumi, Ogai, Midori, Kato, Kosuke, Saito, Haruka, Nishida, Ayano, Sawanaga, Akari, Okada, Sawako, Sakagami, Midori

[International Worm Meeting, 2015]

According to the DOHaD (Developmental Origins of Health and Disease) concept, poor nutritional environment during early life (including conception, embryogenesis, infancy and early childhood) induces changes in development that increase the disease risk in adult stage. The DOHaD is attracting many attention because not only it could change our view toward health and diseases, but also it causes concern about the health of our future generation. Generally, the three molecular events, DNA methylation, histone modifications, and non-coding RNAs are considered to be involved in DOHaD process in vertebrates, but the precise mechanism is not well defined. To gain information concerning the molecular mechanism of DOHaD, starvation was introduced to the parent worms, and the offsprings were analyzed for their fecundity, life span, mobility and fat accumulation. There were no difference observed in fecundity, life span and mobility between the offsprings derived from starvation-experienced and fed-parent worms. However, Nile Red staining showed that fat content was significantly increased in the offsprings derived from starvation-experienced worms compared with that of control offsprings. Fat accumulation was further indicated by gas chromatography analysis. DNA microarray is now being conducted to analyse the gene expression pattern.The data is in line with DOHaD concept, which may indicate that C.elegans can be used in DOHaD model animal.

Amado M et al. (1998) J Biol Chem "A family of human beta3-galactosyltransferases. Characterization of four members of a UDP-galactose:beta-N-acetyl-glucosamine/beta-nacetyl-galactosamine beta-1,3-galactosyltransferase family."

Almeida R, Schwientek T, Amado M, Nielsen PA, Carneiro F, Hollingsworth MA, Nomoto M, Levery SB, Holmes EH, Bennett EP, Clausen H, Hassan H

[J Biol Chem, 1998]

BLAST analysis of expressed sequence tags (ESTs) using the coding sequence of a human UDP-galactose:beta-N-acetyl-glucosamine beta-1, 3-galactosyltransferase, designated beta3Gal-T1, revealed no ESTs with identical sequences but a large number with similarity. Three different sets of overlapping ESTs with sequence similarities to beta3Gal-T1 were compiled, and complete coding regions of these genes were obtained. Expression of two of these genes in the Baculo virus system showed that one represented a UDP-galactose:beta-N-acetyl-glucosamine beta-1, 3-galactosyltransferase (beta3Gal-T2) with similar kinetic properties as beta3Gal-T1. Another gene represented a UDP-galactose:beta-N-acetyl-galactosamine beta-1, 3-galactosyltransferase (beta3Gal-T4) involved in GM1/GD1 ganglioside synthesis, and this gene was highly similar to a recently reported rat GD1 synthase (Miyazaki, H., Fukumoto, S., Okada, M., Hasegawa, T., and Furukawa, K. (1997) J. Biol. Chem. 272, 24794-24799). Northern analysis of mRNA from human organs with the four homologous cDNA revealed different expression patterns. beta3Gal-T1 mRNA was expressed in brain, beta3Gal-T2 was expressed in brain and heart, and beta3Gal-T3 and -T4 were more widely expressed. The coding regions for each of the four genes were contained in single exons. beta3Gal-T2, -T3, and -T4 were localized to 1q31, 3q25, and 6p21.3, respectively, by EST mapping. The results demonstrate the existence of a family of homologous beta3-galactosyltransferase genes.

Tanya Jonassen et al. (2003) International Worm Meeting "Longer is better: an analysis of reproductive fitness and quinone content of C. elegans clk-1 mutants fed coenzyme Q isoforms of varying tail length"

Catherine F Clarke, Tanya Jonassen, Pamela L Larsen, Diana E Davis

[International Worm Meeting, 2003]

C. elegans clk-1 mutants lack coenzyme Q9 and instead accumulate demethoxy-Q9 (DMQ9). When the clk-1 animals are fed a Q-less diet as hatchlings, they arrest as L2 larvae. If the Q-less diet is initiated at a later stage, dauer larvae, then the clk-1 mutants develop into sterile adults. Thus, Q is required for larval growth and development of the gonad and germ cells. These results suggest that DMQ alone cannot functionally replace Q when energy demands are high, and the presence of dietary Q8 prevents the arrest and sterility phenotypes. The Clk-1 phenotypes of slowed development, slowed behaviors, and reduced brood sizes are observed when the animals are fed the Q-replete E. coli. It is not understood why the animals are abnormal when supplemented with the cofactor that they fail to synthesize. It does not appear to be due to lack of assimilation of the bacterial Q, since the E. coli-specific Q8 is present in mitochondria isolated from clk-1(qm30) adults fed a Q-replete diet of OP50. We considered that uptake of the shorter Q8 isoform might contribute to the Clk-1 phenotypes because the worms normally synthesize the Q9 isoform. To test this hypothesis, N2 and clk-1 animals were fed bacteria that had been engineered to produce different isoforms of Q, specifically Q7, Q8, Q9, and Q10 [Okada et al. (1997) J. Bact. 179, 5992; Okada et al. (1995) BBA 1302, 217]. The effects of feeding different Q isoforms on both reproductive fitness and progeny viability were determined. The wild type adults showed no change in reproductive fitness regardless of the food source provided. However, the clk-1 mutants showed a decrease in egg production and decreased viability when they were fed the Q7-containing bacteria. Some of the clk-1(qm30) adults were sterile. Another striking difference was seen on the Q9 and Q10-producing bacteria. Unlike clk-1(qm30), which showed no significant difference compared to OP50-fed animals on these two foods, clk-1(e2519) showed a statistically significant increase in egg production. Uptake of Q7 and Q8 by all genotypes was confirmed by analysis of the quinone profiles. The functional impact of varying the length of the isoprene tail is only observed in animals that cannot synthesize Q and lack functional CLK-1. The impact is negative for the short seven-isoprene tail, whereas the impact is positive for Q9 and Q10 in the weaker e2519 allele. Because the tail length should not alter the redox properties of Q, the differential rescue of fertility by Q isoforms may result from changes in Q localization and/or altered recognition by Q-dependent dehydrogenases.

Campagna S et al. (1996) Midwest Worm Meeting "HUNC-104 IS THE HUMAN COUNTERPART TO UNC-104"

Otsuka AJ, Campagna S

[Midwest Worm Meeting, 1996]

unc-104 is a kinesin-related gene found in the nematode Caenorhabditis elegans and is believed to act as a cytoskeletal motor protein in the nervous system. Kinesin and other motor proteins use ATP hydrolysis to perform microtubule-based transport tasks, such as movement of chromosomes and axonal transport. Motor proteins play a critical role in axonal cell maintenance since many proteins are made in the cell body and must be moved to their sites of utilization. UNC-104 appears to be required for axonal transport of synaptic vesicles in C. elegans and thus is essential for most neural functions (Hall, D.H. and Hedgecock, E.M. 1991. Cell. 65: 837-847.) The murine homolog of unc-104, KIF1A, has also been sequenced and characterized. KIF1A is a neuron-specific anterograde motor for transport of membrane organelles containing synaptic vesicle proteins, such as synaptotagmin and synaptophysin (Okada, Y. et al., 1995. Cell. 81: 769-780). A protein closely related to UNC-104 and KIF1A has now been found in humans and we have sequenced the carboxyl terminal end. Our results show that the translated portion of the sequenced human unc-104 gene (HUNC-104) has 94% amino acid identity with KIF1A and 41% amino acid identity with unc-104. HUNC-104 thus appears to be the human homologue of KIF1A and UNC-104. The results of these experiments may prove useful in treating human nerve disorders, since the HUNC-104 protein is probably involved in neuronal transport. We propose that HUNC-104 is neuron-specific and is involved in transport of synaptic vesicles, similar to UNC-104 and KIF1A

Scholey JM et al. (2000) Midwest Worm Meeting "Characterization of Axonal Transport by UNC-104 in C. elegans"

Zhou HM, Scholey JM

[Midwest Worm Meeting, 2000]

The neuron is a highly polarized cell that elaborates two processes, namely a dendrite and an axon. Dendrites are specialized for neuronal signal reception and axons are specialized for signal conduction and transmission. The formation and function of these processes is thought to depend on the microtubule-based transport of cellular components from their sites of synthesis in the neuronal cell bodies to their sites of utilization at the process ends. We are interested in knowing what kind of motors play roles in such neuronal transport and what is the specific relationship between the motors and the cargoes that they carry along dendrites and axons. It has been reported that UNC-104 is a kinesin motor that is responsible for the anterograde axonal transport of synaptic vesicles (1, 2, 3). Mutations in unc-104 cause synaptic vesicles to accumulate in the cell bodies and the absence of such synaptic vesicles in the presynaptic terminals. Previously our lab has used an in vivo time-lapse fluorescence microscopic transport assay to demonstrate the role of Kinesin II in the anterograde transport of raft particles along the dendrites of C. elegans chemosensory neurons (4, 5). In the current studies, we have used a similar approach to generate transgenic lines that express UNC104::GFP and we are investigating UNC-104 transport along axons. Eventually we want to compare the axonal transport of UNC-104 motor with its cargo (another labeled membrane protein in synaptic vesicles) using the same time-lapse fluorescence microscopic assay. 1.Hall and Hedgecock, 1991 Cell. 65: 837-847 2.Otsuka et al., 1991 Neuron. 6: 113-122 3.Okada et al., 1995 Cell. 81: 769-780 4.Orozco et al., 1999 Nature. 398: 674 5.Signor et al., 1999 Journal of Cell Biology. 147(3): 519-530

T. Schedletzky et al. (2007) International Worm Meeting "Role of the adaptor protein SOC-1 in ligand-gated ion channel clustering at the NMJ."

H. Urlaub, J. Liewald, T. Schedletzky, A. Gottschalk

[International Worm Meeting, 2007]

Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels mediating excitation of muscles and neurons. At the neuromuscular junction (NMJ), nAChRs are clustered opposite of motoneuron release sites, a process regulated in vertebrates by receptor tyrosine kinases (RTKs), e.g. MuSK. We and others showed that in C. elegans, CAM-1 and EGL-15 RTKs affect NMJ clustering of nAChRs (1, 2). Recently, the RTK cytoplasmic adaptor protein Dok-7, that interacts with MuSK, was shown to be essential for nAChR clustering in mice (3). We biochemically co-purified a similar adaptor protein, SOC-1, with the levamisole receptor (LevR; 1). Like Dok-7, SOC-1 has an N-terminal pleckstrin homology (PH) domain and C-terminal SH2 domain binding sites. We found that SOC-1 influences postsynaptic LevR levels, and that soc-1(n1789) mutants are resistant to cholinergic agonists (levamisole and nicotine). In addition, soc-1 mutants were muscimol- (GABA agonist) resistant, and synaptic expression of UNC-49 GABAA receptors was also reduced, indicating functions of SOC-1 beyond nAChR clustering. Cholinergic phenotypes were rescued by expression of SOC-1::GFP in muscles, indicating cell-autonomous function. SOC-1::GFP localized to the plasma membrane, including muscle arms, consistent with its interaction with the LevR. By electrophysiology, soc-1(n1789) mutants showed reduced levamisole- or nicotine-evoked currents in muscle. However, ACh-, as well as GABA- and muscimol-evoked currents were normal. The latter findings could indicate dispersed clustering of GABAARs, rather than reduced surface expression, which we try to confirm by evoking endogenous GABA release using the light-activated cation channel ChR2 (4). In addition, we aim to decode the protein interaction network orchestrating postsynaptic organization, which should include SOC-1 and the LevR. To this end, we purify SOC-1-associated proteins from muscle cells by tandem affinity purification (TAP). Most of the SOC-1::TAP protein is soluble. Initial purification from the soluble fraction yielded a number of proteins which will be identified by mass spectrometry. Furthermore, purification of SOC-1::TAP from the membrane fraction is underway, possibly identifying associated membrane proteins like the LevR, or RTKs acting upstream of SOC-1. Candidate RTKs affecting cholinergic agonist sensitivity were also identified by screening available mutants. 1) Gottschalk et al. (2005) EMBO J 2) Francis et al. (2005) Neuron 3) Okada et al. (2006) Science 4) Nagel et al. (2005) Curr Biol.