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First published online 16 March 2005
doi: 10.1242/dev.01785
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1 Dipartimento di Morfologia Umana e Biologia Applicata, Sezione di Biologia e
Genetica, Università di Pisa, Pisa, Italy
2 Dipartimento di Fisiologia e Biochimica, Laboratorio di Biologia Cellulare e
dello Sviluppo, Università di Pisa, Pisa, Italy
3 Istituto di Fisiologia Clinica, Laboratorio di Terapia Genica e Molecolare,
CNR, Pisa, Italy
4 Dipartimento di Patologia Sperimentale Biotecnologie Mediche, Infettivologia e
Epidemiologia Università di Pisa, Pisa, Italy
Author for correspondence (e-mail:
gremigni{at}biomed.unipi.it)
Accepted 14 February 2005
As stem cells are rare and difficult to study in vivo in adults, the use of classical models of regeneration to address fundamental aspects of the stem cell biology is emerging. Planarian regeneration, which is based upon totipotent stem cells present in the adult – the so-called neoblasts – provides a unique opportunity to study in vivo the molecular program that defines a stem cell. The choice of a stem cell to self-renew or differentiate involves regulatory molecules that also operate as translational repressors, such as members of PUF proteins. In this study, we identified a homologue of the Drosophila PUF gene Pumilio (DjPum) in the planarian Dugesia japonica, with an expression pattern preferentially restricted to neoblasts. Through RNA interference (RNAi), we demonstrate that gene silencing of DjPum dramatically reduces the number of neoblasts, thus supporting the intriguing hypothesis that stem cell maintenance may be an ancestral function of PUF proteins.
Key words: Planarians, Regeneration, Neoblasts, Stem cells, RNAi, Cell proliferation, Pumilio, PUF proteins, Confocal microscopy, TEM, Cytofluorimetry, In situ hybridization, Post-transcriptional regulation
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