Rab8a vesicles regulate wnt ligand delivery and paneth cell maturation at the intestinal stem cell niche

Soumyashree Das, Shiyan Yu, Ryotaro Sakamori, Pavan Vedula, Qiang Feng, Juan Flores, Andrew Hoffman, Jiang Fu, Ewa Stypulkowski, Alexis Rodriguez, Radoslaw Dobrowolski, Akihiro Harada, Wei Hsu, Edward M. Bonder, Michael P. Verzi, Nan Gao

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Communication between stem and niche supporting cells maintains the homeostasis of adult tissues. Wnt signaling is a crucial regulator of the stem cell niche, but the mechanism that governs Wnt ligand delivery in this compartment has not been fully investigated. We identified that Wnt secretion is partly dependent on Rab8amediated anterograde transport of Gpr177 (wntless), a Wnt-specific transmembrane transporter. Gpr177 binds to Rab8a, depletion of which compromises Gpr177 traffic, thereby weakening the secretion of multiple Wnts. Analyses of generic Wnt/β-catenin targets in Rab8a knockout mouse intestinal crypts indicate reduced signaling activities; maturation of Paneth cells – a Wnt-dependent cell type – is severely affected. Rab8a knockout crypts show an expansion of Lgr5+ and Hopx+ cells in vivo. However, in vitro, the knockout enteroids exhibit significantly weakened growth that can be partly restored by exogenous Wnts or Gsk3β inhibitors. Immunogold labeling and surface protein isolation identified decreased plasma membrane localization of Gpr177 in Rab8a knockout Paneth cells and fibroblasts. Upon stimulation by exogenous Wnts, Rab8a-deficient cells show ligand-induced Lrp6 phosphorylation and transcriptional reporter activation. Rab8a thus controls Wnt delivery in producing cells and is crucial for Paneth cell maturation. Our data highlight the profound tissue plasticity that occurs in response to stress induced by depletion of a stem cell niche signal.

Original languageEnglish (US)
Pages (from-to)2147-2162
Number of pages16
JournalDevelopment (Cambridge)
Volume142
Issue number12
DOIs
StatePublished - Jun 15 2015
Externally publishedYes

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Keywords

  • Gpr177
  • Intestinal stem cell
  • Paneth cell
  • Rab8a
  • Wnt secretion
  • Wntless

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology

Cite this

Das, S., Yu, S., Sakamori, R., Vedula, P., Feng, Q., Flores, J., Hoffman, A., Fu, J., Stypulkowski, E., Rodriguez, A., Dobrowolski, R., Harada, A., Hsu, W., Bonder, E. M., Verzi, M. P., & Gao, N. (2015). Rab8a vesicles regulate wnt ligand delivery and paneth cell maturation at the intestinal stem cell niche. Development (Cambridge), 142(12), 2147-2162. https://doi.org/10.1242/dev.121046