Epiblast integrity requires CLASP and Dystroglycan-mediated microtubule anchoring to the basal cortex.
Amniote epiblast cells differentiate into mesoderm and endoderm lineages during gastrulation through a process called epithelial-to-mesenchymal transition (EMT). Molecular regulation of gastrulation EMT is poorly understood. Here we show that epiblast epithelial status was maintained by anchoring microtubules to the basal cortex via CLIP-associated protein (CLASP), a microtubule plus-end tracking protein, and Dystroglycan, a transmembrane protein that bridges the cytoskeleton and basement membrane (BM). Mesoderm formation required down-regulation of CLASP and Dystroglycan, and reducing CLASP activity in pregastrulation epiblast cells caused ectopic BM breakdown and disrupted epiblast integrity. These effects were mediated through the CLASP-binding partner LL5. Live-imaging using EB1-enhanced GFP (eGFP) revealed that reducing CLASP and LL5 levels in the epiblast destabilized basal microtubules. We further show that Dystroglycan is localized to basolateral membrane in epiblast cells. Basal but not lateral localization of Dystroglycan was regulated by CLASP. We propose that epiblast-BM interaction requires CLASP- and Dystroglycan-mediated cortical microtubule anchoring, the disruption of which initiates gastrulation EMT.
Laboratory for Early Embryogenesis, Institute of Physical and Chemical Research RIKEN Center for Developmental Biology, Chuo-Ku Kobe, Hyogo 650-0047, Japan.
Yukiko
Y
Nakaya
Erike W
EW
Sukowati
Guojun
G
Sheng
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism*; D002645:Chickens; D049030:Dystroglycans / Q000378:metabolism*; D005855:Germ Layers / Q000166:cytology / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data
C497850:CLASP1 protein, human; C497851:CLASP2 protein, human; D008869:Microtubule-Associated Proteins; D049030:Dystroglycans