Production and characterisation of cell- and tissue-specific monoclonal antibodies for the flatworm Macrostomum sp.

Monoclonal antibodies (mABs) against various cell types of the basal free-living flatworm Macrostomum sp. were produced by immunising Balb/c mice with cell suspensions of disintegrated animals. We identified 360 positive supernatants with specific staining of various tissues, cell types, patterns or structures. Here we report immunocytochemical characterisation, histological stainings and isotyping of 11 mABs specific for muscle cells (MMu-1, MMu-2, MMu-3, MMu-4), digestive and prostate glands (MDr-1 and MDr-2, MPr-1), epidermal cells (MEp-1), the ventral nerve cord including neuron clusters (MNv-1), gastrodermal cells (MDa-1) and spermatids (MSp-1). Confocal microscopy, histological techniques, electron microscopy and immunoblotting were applied to demonstrate stainings in juveniles, adults, starved or well-fed animals. Considering the current lack of specific markers the obtained mABs will be particularly helpful studying embryonic and postembryonic development, pattern formation, cell differentiation, regeneration and reproductive allocation in Macrostomum sp., and possibly other basal flatworms. The small size, ease of culturing, short generation time, transparency and the basal phylogenetic position specify Macrostomum sp. as a suitable model organism for comparative analyses within Platyhelminthes and to Drosophila and C. elegans.

Stem cell dynamics during growth, feeding, and starvation in the basal flatworm Macrostomum sp. (Platyhelminthes).

Development, growth, and regeneration in Macrostomum are based--as in all Platyhelminthes--on likely totipotent stem cells (neoblasts), basic for all Bilaterians. We demonstrate dynamics and migration of neoblasts during postembryonic development, starvation, and feeding of Macrostomum sp. Double labeling of S-phase and mitotic cells revealed a fast cell turnover. Conflicting with recent results from planarians, we have some indication of slow cycling neoblasts. As in planarians, starvation dramatically reduced mitotic activity and a very basic level was maintained after 30 days of starvation. Afterward, feeding induced a dramatic immediate proliferative response probably caused by G2-arrested neoblasts. The following 12 hr showed a significant mitotic decline, caused by the depletion of the G2 neoblast pool. Neoblasts that pass through S-phase led to a maximum of mitoses after 48 hr. Our results allow deeper insight into cellular dynamics of an ancestral bilaterian stem cell system of a basal Platyhelminth.