TY - JOUR
T1 - An RNAi Screen Reveals Intestinal Regulators of Branching Morphogenesis, Differentiation, and Stem Cell Proliferation in Planarians
AU - Forsthoefel, David J.
AU - James, Noëlle P.
AU - Escobar, David J.
AU - Stary, Joel M.
AU - Vieira, Ana P.
AU - Waters, Forrest A.
AU - Newmark, Phillip A.
N1 - Funding Information:
We thank John Brubacher, Jim Collins, Tingxia Guo, Ryan King, Melanie Issigonis, Rachel Roberts-Galbraith, Labib Rouhana, James Sikes, Yuying Wang, Ricardo Zayas, and other Newmark laboratory members for their insights, comments, and discussions; Tracy Chong for collaboration on dispase dissociation; Ryan King for advice and collaboration on FISH optimization; David Horn and Doug LaCount for kindly sharing p2T7TAblue and other trypanosome dsRNA expression vectors; Liping Wang, Rachel Breitenfeld, and Xiaoxia Wang of the Immunological Resources Center at Illinois for monoclonal antibody production; Jenny Drnevich Zadeh for guidance in microarray analysis; Barbara Pilas for suggesting magnetic sorting; and Diana Thomas for helping with magnetic sorting. We apologize to authors whose work we were unable to cite due to space constraints. This work was supported by a Ruth L. Kirschstein National Research Service Award from the National Institutes of Health (F32-DK077469) to D.J.F. and R01-HD043403 to P.A.N. P.A.N. is an investigator of the Howard Hughes Medical Institute.
PY - 2012/10/16
Y1 - 2012/10/16
N2 - Planarians grow and regenerate organs by coordinating proliferation and differentiation of pluripotent stem cells with remodeling of postmitotic tissues. Understanding how these processes are orchestrated requires characterizing cell-type-specific gene expression programs and their regulation during regeneration and homeostasis. To this end, we analyzed the expression profile of planarian intestinal phagocytes, cells responsible for digestion and nutrient storage/distribution. Utilizing RNA interference, we identified cytoskeletal regulators required for intestinal branching morphogenesis and a modulator of bioactive sphingolipid metabolism, ceramide synthase, required for the production of functional phagocytes. Additionally, we found that a gut-enriched homeobox transcription factor, nkx-2.2, is required for somatic stem cell proliferation, suggesting a niche-like role for phagocytes. Identification of evolutionarily conserved regulators of intestinal branching, differentiation, and stem cell dynamics demonstrates the utility of the planarian digestive system as a model for elucidating the mechanisms controlling postembryonic organogenesis. Organ regeneration coordinates stem cell proliferation and differentiation with remodeling of postmitotic tissues. Forsthoefel et al. purify planarian intestinal cells, characterize their expression profile, and perform a targeted RNAi screen. Their work on transcription factor Nkx2.2 suggests that intestinal phagocytes play a niche-like role to maintain neighboring pluripotent stem cells.
AB - Planarians grow and regenerate organs by coordinating proliferation and differentiation of pluripotent stem cells with remodeling of postmitotic tissues. Understanding how these processes are orchestrated requires characterizing cell-type-specific gene expression programs and their regulation during regeneration and homeostasis. To this end, we analyzed the expression profile of planarian intestinal phagocytes, cells responsible for digestion and nutrient storage/distribution. Utilizing RNA interference, we identified cytoskeletal regulators required for intestinal branching morphogenesis and a modulator of bioactive sphingolipid metabolism, ceramide synthase, required for the production of functional phagocytes. Additionally, we found that a gut-enriched homeobox transcription factor, nkx-2.2, is required for somatic stem cell proliferation, suggesting a niche-like role for phagocytes. Identification of evolutionarily conserved regulators of intestinal branching, differentiation, and stem cell dynamics demonstrates the utility of the planarian digestive system as a model for elucidating the mechanisms controlling postembryonic organogenesis. Organ regeneration coordinates stem cell proliferation and differentiation with remodeling of postmitotic tissues. Forsthoefel et al. purify planarian intestinal cells, characterize their expression profile, and perform a targeted RNAi screen. Their work on transcription factor Nkx2.2 suggests that intestinal phagocytes play a niche-like role to maintain neighboring pluripotent stem cells.
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U2 - 10.1016/j.devcel.2012.09.008
DO - 10.1016/j.devcel.2012.09.008
M3 - Article
C2 - 23079596
AN - SCOPUS:84867686201
VL - 23
SP - 691
EP - 704
JO - Developmental Cell
JF - Developmental Cell
SN - 1534-5807
IS - 4
ER -