Much work has gone into understanding anhydrobiosis at both the whole organism level and the molecular level, but little has been done to understand what happens to the cells of metazoan anhydrobiotes during drying. Work is therefore in progress in the lab to develop a nematode cell model of anhydrobiosis.
A technique has been developed for the production of primary embryonic cell cultures from the model nematode Caenorhabditis elegans (Bloom, 1993; Christensen et al., 2002) and later adapted for Brugia malayi (Higazi et al., 2004). This technique has been applied to the anhydrobiotic nematodes Panagrolaimus superbus and Aphelenchus avenae to produce the first cell cultures from these animals. Live/dead staining (Fig. 1; Fig. 2), where green fluorescence indicates a live cell and red fluorescence a dead cell, can be used to monitor the longevity, and resistance to stress, of cell cultures. C. elegans cell cultures have been shown to differentiate in culture into nerve and muscle cells. Similarly, both P. superbus and A. avenae cells differentiate in culture (Fig. 3) and produce neuron-like processes. Work is currently underway to assess the ability of nematode cell cultures to resist desiccation.
|Fig. 1: Live/dead staining of A. avenae cell cultures.||Fig. 2: Live/dead staining of P. superbus cell cultures.||Fig. 3: A. avenae cells differentiating in culture|
Nematode Research Topics:
Bloom, L. (1993) Genetic and molecular analysis of genes required for axon outgrowth in Caenorhabditis elegans. Ph.D thesis, Massachusetts Institute of Technology, Cambridge, Massachusetts.
Christensen, M., Estevez, A., Yin, X., Fox, R., Morrison, R., McDonnell, M., Gleason, C., Miller, D. M. 3rd. and Strange, K. (2002) A primary culture system for functional analysis of C. elegans neurons and muscle cells. Neuron 33: 503-514.
Higazi, T, .B. Shu, L. and Unnasch, T. R. (2004) Development and transfection of short-term primary cell cultures from Brugia malayi. Mol. Biochem. Parasit. 137: 345-348.