The use of fluorescent marker dyes for studying intercellular communication in nematode embryos

Abstract

As more and more cases of necessary cell-cell interactions are revealed, the classical view of mosaic development in nematodes has to be replaced by a more dynamic picture showing different types of intercellular communication. To investigate the pattern and function of communication pathways between cells, we have developed different techniques to shunt fluorescent marker dyes into embryos and hatched animals and study their distribution in vivo. During embryogenesis we find that for a long time all somatic cells form a single dye-coupling compartment while transfer into the germline is restricted already at an early stage. Considerable variations between species with respect to the size of communication channels and the time during which these are functional are observed and can be correlated to differences in the developmental program. A different kind of intracellular communication can be visualized with the help of fluorescent dyes: a transfer of yolk proteins in two phases of the life cycle, in the adult hermaphrodite from the gut into the maturing germ cells, and in the embryo from non-gut cells into the gut primordium. Cell-cell interactions in the nematode embryo can be inhibited with polysulfated hydrocarbon dyes (e.g. Trypan Blue) which bind strongly to the plasma membrane. In summary our data indicate that fluorescent marker dyes can be helpful tools to identify and understand the role of intercellular communication and transfer processes in nematode development.