Miquel Vila-Farré

Reactivating head regrowth in a regeneration-deficient planarian species

Species capable of regenerating lost body parts occur throughout the animal kingdom, yet close relatives are often regeneration incompetent. Why in the face of ‘survival of the fittest’ some animals regenerate but others do not remains a fascinating question. Planarian flatworms are well known and studied for their ability to regenerate from minute tissue pieces, yet species with limited regeneration abilities have been described even amongst planarians. Here we report the characterization of the regeneration defect in the planarian Dendrocoelum lacteum and its successful rescue. Tissue fragments cut from the posterior half of the body of this species are unable to regenerate a head and ultimately die. We find that this defect originates during the early stages of head specification, which require inhibition of canonical Wnt signalling in other planarian species. Notably, RNA interference (RNAi)-mediated knockdown of Dlac-β-catenin-1, the Wnt signal transducer, restored the regeneration of fully functional heads on tail pieces, rescuing D. lacteum’s regeneration defect. Our results demonstrate the utility of comparative studies towards the reactivation of regenerative abilities in regeneration-deficient animals. Furthermore, the availability of D. lacteum as a regeneration-impaired planarian model species provides a first step towards elucidating the evolutionary mechanisms that ultimately determine why some animals regenerate and others do not.

PlanMine – a mineable resource of planarian biology and biodiversity

Planarian flatworms are in the midst of a renaissance as a model system for regeneration and stem cells. Besides two well-studied model species, hundreds of species exist worldwide that present a fascinating diversity of regenerative abilities, tissue turnover rates, reproductive strategies and other life history traits. PlanMine (http://planmine.mpi-cbg.de/) aims to accomplish two primary missions: First, to provide an easily accessible platform for sharing, comparing and value-added mining of planarian sequence data. Second, to catalyze the comparative analysis of the phenotypic diversity amongst planarian species. Currently, PlanMine houses transcriptomes independently assembled by our lab and community contributors. Detailed assembly/annotation statistics, a custom-developed BLAST viewer and easy export options enable comparisons at the contig and assembly level. Consistent annotation of all transcriptomes by an automated pipeline, the integration of published gene expression information and inter-relational query tools provide opportunities for mining planarian gene sequences and functions. For inter-species comparisons, we include transcriptomes of, so far, six planarian species, along with images, expert-curated information on their biology and pre-calculated cross-species sequence homologies. PlanMine is based on the popular InterMine system in order to make the rich biology of planarians accessible to the general life sciences research community.

Evidence for the persistence of the land planarian species Microplana terrestris (Müller, 1774) (Platyhelminthes, Tricladida) in microrefugia during the Last Glacial Maximum in the northern section of the Iberian Peninsula

The land planarian species Microplana terrestris (Müller, 1774), shows a wide distribution in the north of the Iberian Peninsula, where mature humid forests can be found. Since most terrestrial planarians require the presence and good condition of wet forests to survive, a parallel evolution of the taxon and its habitat might be expected. Performing molecular analyses (mitochondrial cytochrome oxidase I and nuclear ITS-1 genes) we estimated the demography and biogeographic history of the species in that region. Our results show the species to present levels of genetic diversity likely originating before the Pleistocene. However, it presents a genetic structure that presumably resulted from its survival in various refugees during the Pleistocene glacial cycles. The two main genetic groups, present on the Iberian Peninsula, seem to have different origins: the western one being of Iberian origin, while the eastern group may have been the result of a re-colonization from the north. In both cases, their biogeographical history mirrors their habitat range movements, reinforcing the phylogeographical hypothesis put forward for its preferred habitat, i.e. humid forests.

The diet and distribution of Pentacoelum kazukolinda (Kawakatsu & Mitchell, 1984), a maricolan planarian with a freshwater ecology

Among the marine triclads or Maricola, the genus Pentacoelum represents one of the few exceptions in that species occur in brackish water or in freshwater. We examined specimens of a maricolan triclad that externally looked remarkably similar to the continental Spanish representatives of freshwater Pentacoelum hispaniense, albeit that these new samples came from habitats in Germany, the Netherlands, Mallorca and Japan. Comprehensive comparative morphological studies revealed that the animals from these populations are identical to a species described earlier from Hawaii, viz. Oahuhawaiiana kazukolinda Kawakatsu & Mitchell, 1984. It is argued that the species now should be called Pentacoelum kazukolinda (Kawakatsu & Mitchell, 1984) comb. nov. Remarkably, Dutch and German specimens of P. kazukolinda almost completely lack any signs of the presence of the lateral gonopore system, being the defining feature of their taxonomic family, the Bdellouridae. The reason for this reduction or even complete absence of this structure remains obscure. Nonetheless, morphological similarities and 18S rDNA sequences strongly suggest that all animals belong to the same species. Analyses of the mitochondrial gene sequences Cox1 and 16SrDNA of the gut content of P. kazukolinda revealed that the flatworm has a preference for different species of gastropods, while in one population annelids were detected. Our study suggests that gastropods may form the food refuge of P. kazukolinda. Circumstantial evidence suggests that the worldwide distribution of P. kazukolinda may result from human‐mediated transport of the aroid Colocasia esculenta and/or the introduced snail Potamopyrgus antipodarum. The use of P. antipodarum as a food resource may have facilitated the spread and establishment of new populations of P. kazukolinda.

Marine planarians (Platyhelminthes: Tricladida: Maricola) from the western Mediterranean Sea and the Cantabrian coast: new records, one new genus, and immunocytochemistry of the nervous system

Marine planarians were collected extensively from the Iberian Peninsula and Italy. As a result we provide new distributional records of six species of marine triclads, including the description of one new genus and species. The study increases substantially our knowledge of the distribution of this group of animals in Spain and reveals that even relatively well studied areas, such as Italy, still yield new species. In addition, we performed immunostaining studies of the nervous system of three of the species, which revealed the detailed organization of the main nerve cords and their branches. In the new species, the lateral nerve branches showed an arrangement that is different from that of the other species.

The Ecology of Freshwater Planarians

Planarians are on the rise as a model system for regeneration and stem cell dynamics. Almost in parallel the interest in planarian field biology has declined. Besides representing an independent research discipline in its own right, understanding of the natural habitat is also directly relevant to optimizing culture conditions in the laboratory. Moreover, the current laboratory models are but few of hundreds of planarian species worldwide. Their adaptation to a wide range of ecological niches has resulted in a fascinating diversity of regenerative abilities, body size, reproduction strategies, and life expectancy, to name just a few. With the currently ongoing establishment of large planarian species collections, such phenotypic diversity becomes accessible to comparative mechanistic analysis in the laboratory. Overall, we hope that this chapter inspires an integral view of the planarian model system that not only includes the molecular and cellular processes under investigation but also the evolutionary forces that shaped them in the first place.

Small- and Large-Scale High Molecular Weight Genomic DNA Extraction from Planarians

High-quality genomic DNA extraction is a starting point for many downstream applications in modern molecular biology. Here, we describe a simple method for isolating high molecular weight genomic DNA from planarians. The method is based on tissue lysis by a mixture of a chaotropic salt and detergent followed by organic extraction to remove proteins and lipids followed by a postpurification step to remove contaminating polysaccharides. The isolated DNA is of high molecular weight and compatible with polymerase chain reaction, cloning, or next-generation sequencing library preparation.