Glypican 4 and Mmp14 interact in regulating the migration of anterior endodermal cells by limiting extracellular matrix deposition

Abstract

During embryogenesis, the germ layers, including the endoderm, undergo convergence and extension movements to narrow and elongate the body plan. In zebrafish, the dorsal migration of endodermal cells during gastrulation is controlled by chemokine signaling, but little is known about how they migrate during segmentation. Here, we show that glypican 4 (Gpc4), a member of the heparin sulfate proteoglycan family, is required for efficient migration of anterior endodermal cells during early segmentation, regulating Rac activation to maintain polarized actin-rich lamellipodia. An endoderm transplantation assay showed that Gpc4 regulates endoderm migration in a non-cell-autonomous fashion. Further analyses revealed that the impaired endoderm migration in mutants results from increases in the expression and assembly of fibronectin and laminin, major components of the extracellular matrix (ECM). Notably, we found that matrix metalloproteinase 14 (Mmp14a/b) is required for the control of ECM expression during endoderm migration, with Gpc4 acting through Mmp14a/b to limit ECM expression. Our results suggest that Gpc4 is crucial for generating the environment required for efficient migration of endodermal cells, uncovering a novel function of Gpc4 during development.

Glypican4 mediates Wnt transport between germ layers via signaling filopodia

“…Previously, we detected gpc4 transcript in the anterior endoderm (Hu, et al, 2018). Here, we found that gpc4 is also expressed in the posterior endoderm ( Fig.…”

“…S1D). GFP-Gpc4 is functional, as injection of the encoding RNA rescued C&E defects in gpc4 mutants (Hu, et al, 2018). Consistent with the expression pattern of sox17 (Alexander, et al, 1999), in this line GFP-Gpc4 signal was detected in the innermost tissue of the embryo, including in Kupffer's vesicle at the tailbud (TB) stage .…”

“…In humans, mutations in the GPC3, GPC4 and GPC6 genes are associated with congenital diseases such as Simpson-Golabi-Behmel (SGB) overgrowth syndrome (Campos-Xavier, et al, 2009;DeBaun, et al, 2001;Veugelers, et al, 1998;Pilia, et al, 1996). Studies in various animal models have revealed that Gpcs play roles in various developmental processes including: wing and eye development; the establishment of body length; jaw and heart development; and the migration of endodermal cells and cells of the lateral line primordium (LLP) (Hu, et al, 2018;Miles, et al, 2017;Venero Galanternik, et al, 2016;Strate, et al, 2015;Zhang, et al, 2013;LeClair, et al, 2009;Gumienny, et al, 2007).…”

“…Recently, we and others showed that Gpc4 is required for endoderm C&E, in both the anterior and posterior regions (Hu, et al, 2018;Miles, et al, 2017). To investigate the cell autonomy of Gpc4 in the gut endoderm, we generated a transgenic line that expresses Gpc4 specifically in this germ layer.…”

CARMIL3 is important for cell migration and morphogenesis during early development in zebrafish

“…Whole-mount RNA in situ hybridization (WISH) Digoxigenin-labeled antisense RNA probes for myl7 (cardiac myosin regulatory light chain) (Ye and Lin, 2013;Yelon et al, 1999), sox17 (sex determining region Y-box 17) (Alexander et al, 1999;Hu et al, 2018), southpaw (spaw) (Long et al, 2003;Panizzi et al, 2007) were synthesized by in vitro transcription. Staged embryos were fixed in 4% fish fix solution (4% paraformaldehyde, 4% sucrose, 0.1 M phosphate buffer pH 7.2, 0.12 mM CaCl2) at 4˚C overnight.…”