Keiji Fukui et al. (2005) International Worm Meeting "Checkpoint and physiological apoptosis in germ cells proceeds normally in spaceflown Caenorhabditis elegans on the ICE first experiment."

Michel Viso, Gilbert Gasset, Yutaka Miyazawa, Brigitte Eche, Atsushi Higashitani, Yohei Sasagawa, Nathaniel Szewczyk, Tomoko Sugimoto, Keiji Fukui, Akira Higashibata

[International Worm Meeting, 2005]

It is important for human life in space to study the effects of environmental factors during spaceflight on a number of physiological phenomena. Apoptosis plays important roles in development and tissue homeostasis in metazoans. In this study, we have analyzed apoptotic activity in germ cells of the nematode C. elegans, following spacefight. Comparison of the number of cell corpses in wild type or ced-1 mutants, grown under either ground or spaceflight conditions, showed that both pachytene-checkpoint apoptosis and physiological apoptosis in germ cells occurred normally under spaceflight conditions. In addition, the expression levels of the checkpoint and apoptosis related genes are comparable between spaceflight and ground conditions. This is the first report documenting the occurrence of checkpoint apoptosis in the space environment and suggests that metazoans, including humans, would be able to eliminate cells that have failed to repair DNA lesions introduced by cosmic radiation during spaceflight.

Nobuyoshi Fujimoto et al. (2005) International Worm Meeting "Microgravity-dependent alteration of protein and gene expression in Caenorhabditis elegans on the international C.elegans space experiment-first (ICE-first)"

Nobuyoshi Fujimoto, Naoko Higashitani, Noriaki Ishioka, Michel Viso, Akira Higashibata, Mari Sato, Nathaniel Szewczyk, Brigitte Eche, Masaya Seki, Gilbert Gaset

[International Worm Meeting, 2005]

We have participated in the international C.elegans space experiment-first (ICE-First) that is a cooperative project of JAXA and CNES with NASA, CSA, and ESA using the nematode C.elegans. The protein alteration of C.elegans cultured in the international space station (ISS) was analyzed by combined method with two-dimensional gel electrophoresis and mass spectrometry. More than 1268 protein spots in mix stages of C.elegans were detected in SYPRO-Ruby staining gel by computer-assisted image analysis. Comparison of total proteins between the flight and ground groups showed that total 316 protein spots changed on the map of the flight sample, and 254 spots changed more than 2-fold up (167) and down (87). Phosphorylated proteins were also analyzed by staining with ProQ-Diamond and total 198 protein spots were indicated on the gel. In the fright worms, 94 spots changed more than 2-fold up (67) and down (27) as compare to the ground control. Two protein spots that significantly decreased in the flight worms were identified as troponin T by mass spectrometric protein identification, and this was supported by gene expression analysis. In addition, gene expression analysis was focused on G-protein related-proteins including Rho family GTPases and guanine nucleotide exchange factors using the DNA microarray. RhoGEF family such as unc-73B and RhoGAP tended to decrease, and by contraries RhoA increased in the flight animals. These results suggest that the space environment, microgravity, affects networks of intracellular signaling proteins that regulate a variety of cellular event, including cytoskeletal formation and neural development.

Julia Grabitzki et al. (2006) European Worm Meeting "Identification of O-linked N-acetylglucosamine Modified Proteins in Caenorhabditis elegans"

Gunter Lochnit, Rudolf Geyer, Michael Ahrend, Brigitte Schmitz, Julia Grabitzki

[European Worm Meeting, 2006]

Julia Grabitzki1, Michael Ahrend2, Brigitte Schmitz2, Rudolf Geyer1 and Gnter Lochnit1. The posttranslational modification N-acetylglucosamine O-glycosidically linked (O-GlcNAc) to serine and threonine residues of proteins has been shown to be ubiquitous amongst eukaryotic proteins of the nucleus, cytoskeleton, cytoplasm, and has also been detected on cytosolic tails of membrane proteins [1]. O-GlcNAcylated proteins can form reversible multimeric complexes with other polypeptides or structures. The modification is often accompanied by phosphorylation/ dephosphorylation. O-GlcNAc can act either simultaneously or in a reciprocal fashion with phosphorylation. According to the Yin-Yang hypothesis, the phosphorylation/ dephosphorylation regulates O-GlcNAc-modified protein function (z.B. signal transduction and protein-protein interaction) in concert with phosphorylation [2-4]. The addition of O-GlcNAc to and the removal from the protein backbone is dynamic with rapid cycling in response to cellular signals or cellular stages.. Despite the fact, that Caenorhabiditis elegans is the best studied model organism, there have been no studies on O-GlcNAcylation in this organism so far. Therefore, to elucidate the role of O-GlcNAcylation, we investigated the proteome of a C. elegans mixed-stage population by two-dimensional gelelectrophoresis and subsequent western-blotting with the O-GlcNAc-specific antibody CTD 110.6 for the occurrence of this modification and identified the modified proteins by mass-spectrometry. We detected and identify several O-GlcNAc-modified proteins in C. elegans. Most of the identified proteins are involved in metabolic pathways. The prediction of the cellular localisation of the identified proteins revealed a predominant cytosolic occurrence of the O-GlcNAc modification.. References:. [1]. Rex-Mathes, M., J. Koch, Werner, S., Griffith, L. S and B. Schmitz. 2002. Methods Mol Biol 194: 73-87.. [2] Zachara, N.E. and G.W. Hart, Chem Rev, 2002. 102(2): p.431-8.. [3]. Griffith, L. S. and B. Schmitz. 1999. Eur J Biochem 262(3): 824-31.. [4] Wells, L. and G. W. Hart. 2003. FEBS Lett 546(1): 154-8.