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Stem cells (neoblasts) in Platyhelminthes are pluripotent, and likely totipotent, undifferentiated cells which retain throughout adult life the capacity to proliferate and from which all somatic cells as well as the germ cells derive. However, basic data on the pool and heterogeneity of neoblasts, their rates of differentiation into sets and subsets of differentiated cells, and their migration to different body regions are still lacking. To fill this gap, S-phase cells in the macrostomid Macrostomum sp. were labeled with the thymidine analog 5-bromo-2′-deoxyuridine (BrdU). S-phase cells were found to be neoblasts and to be distributed in two bands along the lateral sides of the body leaving unlabeled the median axis of the body and the region anterior to the eyes. This distribution is parallel to that of mitotic cells demonstrated using an antibody to phosphorylated histone H3. At different chase times, clusters of BrdU-labeled cells appear, labeled cells migrate to formerly unlabeled areas, and they differentiate into several somatic cell types and into germ cells. Finally, continuous exposure to BrdU shows an extensive renewal of the epithelial cells. Altogether, these results strengthen the idea of platyhelminth neoblasts as an unparalleled stem-cell system within the Animal Kingdom calling for further investigation.

Sex allocation theory for simultaneous hermaphrodites predicts an influence of the mating group size on sex allocation. Mating group size may depend on the size of the group in which an individual lives, or on the density, but studies to date have not distinguished between the two factors. We performed an experiment in which we raised a transparent simultaneous hermaphrodite, the flatworm Macrostomum sp., in different group sizes (pairs, triplets, quartets and octets) and in different enclosure sizes (small and large). This design allows us to differentiate between the effects of group size and density. After worms reached maturity we determined their reproductive allocation patterns from microscopic images taken in vivo. The results suggest that the mating group size is a function of the group size, and not of the density. They support the shift to higher male allocation in larger mating groups predicted by sex allocation theory. To our knowledge, this is the first study that unambiguously shows phenotypically plastic sex allocation in response to mating group size in a simultaneous hermaphrodite.

The lion’s share of studies on regeneration in Plathelminthes (flatworms) has been so far carried out on a derived taxon of rhabditophorans, the freshwater planarians (Tricladida), and has shown this group’s outstanding regeneration capabilities in detail. Sharing a likely totipotent stem cell system, many other flatworm taxa are capable of regeneration as well. In this paper, we present the regeneration capacity of Macrostomum lignano, a representative of the Macrostomorpha, the basal-most taxon of rhabditophoran flatworms and one of the most basal extant bilaterian protostomes. Amputated or incised transversally, obliquely, and longitudinally at various cutting levels, M. lignano is able to regenerate the anterior-most body part (the rostrum) and any part posterior of the pharynx, but cannot regenerate a head. Repeated regeneration was observed for 29 successive amputations over a period of almost 12 months. Besides adults, also first-day hatchlings and older juveniles were shown to regenerate after transversal cutting. The minimum number of cells required for regeneration in adults (with a total of 25,000 cells) is 4,000, including 160 neoblasts. In hatchlings only 1,500 cells, including 50 neoblasts, are needed for regeneration. The life span of untreated M. lignano was determined to be about 10 months.

Significance: The availability of high-quality genome and transcriptome assemblies is critical for enabling full exploitation of any model organism. Here we present genome and transcriptome assemblies for Macrostomum lignano , a free-living flatworm that can regenerate nearly its entire body following injury. The resources we present here will promote not only the studies of mechanisms of stem cell self-renewal, but also of regeneration and differentiation.