Identification of TOR-responsive slow-cycling neoblasts in planarians.

Epimorphic regeneration commonly relies on the activation of reserved stem cells to drive new cell production. The planarian Schmidtea mediterranea is among the best regenerators in nature, thanks to its large population of adult stem cells, called neoblasts. While neoblasts have long been known to drive regeneration, whether a subset of neoblasts is reserved for this purpose is unknown. Here, we revisit the idea of reserved neoblasts by approaching neoblast heterogeneity from a regulatory perspective. By implementing a new fluorescence-activated cell sorting strategy in planarians, we identify a population of neoblasts defined by low transcriptional activity. These RNAlow neoblasts are relatively slow-cycling at homeostasis and undergo a morphological regeneration response characterized by cell growth at 48 h post-amputation. At this time, RNAlow neoblasts proliferate in a TOR-dependent manner. Additionally, knockdown of the tumour suppressor Lrig-1, which is enriched in RNAlow neoblasts, results in RNAlow neoblast growth and hyperproliferation at homeostasis, and ultimately delays regeneration. We propose that slow-cycling RNAlow neoblasts represent a regeneration-reserved neoblast population.

Myths vs. FACS: what do we know about planarian stem cell lineages?

Historically, planarian neoblasts were thought to be a homogeneous population of pluripotent stem cells; however, recent population and single-cell level analyses have refuted this idea. Evidence for lineage commitment at the neoblast level has been provided via a number of independent studies using a variety of methods. In situ hybridization experiments first demonstrated the co-expression of lineage-specific markers in neoblasts (marked by piwi-1 expression) isolated by FACS. Subsequently, single cell transcriptomic analyses of FACS-isolated neoblasts uncovered broad lineage-primed neoblast classes based on the clustering of transcriptional profiles and expression of known tissue-specific markers. Additionally, single neoblast pluripotency (and fate restriction) has been demonstrated by single cell transplantation experiments into neoblast-void animals. Here we look to recount the current status of the planarian neoblast field and offer discussion on the caveats of neoblast biology and how to address them experimentally.

FOX and ETS family transcription factors regulate the pigment cell lineage in planarians.

Many pigment cells acquire unique structural properties and gene expression profiles during animal development. The underlying differentiation pathways have been well characterized in cells formed during embryogenesis, such as the neural crest-derived melanocyte. However, much less is known about the developmental origins of pigment cells produced in adult organisms during tissue homeostasis and repair. Here we report a lineage analysis of ommochrome- and porphyrin-producing cells in the brown, freshwater planarian Schmidtea mediterranea Using an RNA-sequencing approach, we identified two classes of markers expressed in sequential fashion when new pigment cells are generated during regeneration or in response to pigment cell ablation. We also report roles for FOXF-1 and ETS-1 transcription factors, as well as for an FGFR-like molecule, in the specification and maintenance of this cell type. Together, our results provide insights into mechanisms of adult pigment cell development in the strikingly colorful Platyhelminthes phylum.