TY - JOUR
T1 - Reck and Gpr124 Are Essential Receptor Cofactors for Wnt7a/Wnt7b-Specific Signaling in Mammalian CNS Angiogenesis and Blood-Brain Barrier Regulation
AU - Cho, Chris
AU - Smallwood, Philip M.
AU - Nathans, Jeremy
N1 - Funding Information:
The authors would like to thank Dr. Makoto Noda (Kyoto University) for the Reck flex2 mice, Dr. Chiaki Takahashi (Kanazawa University) and Dr. Shan Sockanathan (Johns Hopkins University) for the Reck Δex1 mice, Dr. Thomas Spencer (University of Missouri) for the Wnt7a fl mice, Dr. Maketo Taketo (Kyoto University) for the Ctnnb1 flex3 mice, the Johns Hopkins University School of Medicine Transgenic Core Laboratory for blastocyst and egg injections, and Mark Sabbagh for helpful comments on the manuscript. This study was supported by the Howard Hughes Medical Institute , the National Eye Institute ( R01 EY018637 ), and the Arnold and Mabel Beckman Foundation .
Publisher Copyright:
© 2017 Elsevier Inc.
PY - 2017/8/30
Y1 - 2017/8/30
N2 - Reck, a GPI-anchored membrane protein, and Gpr124, an orphan GPCR, have been implicated in Wnt7a/Wnt7b signaling in the CNS vasculature. We show here that vascular endothelial cell (EC)-specific reduction in Reck impairs CNS angiogenesis and that EC-specific postnatal loss of Reck, combined with loss of Norrin, impairs blood-brain barrier (BBB) maintenance. The most N-terminal domain of Reck binds to the leucine-rich repeat (LRR) and immunoglobulin (Ig) domains of Gpr124, and weakening this interaction by targeted mutagenesis reduces Reck/Gpr124 stimulation of Wnt7a signaling in cell culture and impairs CNS angiogenesis. Finally, a soluble Gpr124(LRR-Ig) probe binds to cells expressing Frizzled, Wnt7a or Wnt7b, and Reck, and a soluble Reck(CC1-5) probe binds to cells expressing Frizzled, Wnt7a or Wnt7b, and Gpr124. These experiments indicate that Reck and Gpr124 are part of the cell surface protein complex that transduces Wnt7a- and Wnt7b-specific signals in mammalian CNS ECs to promote angiogenesis and regulate the BBB.
AB - Reck, a GPI-anchored membrane protein, and Gpr124, an orphan GPCR, have been implicated in Wnt7a/Wnt7b signaling in the CNS vasculature. We show here that vascular endothelial cell (EC)-specific reduction in Reck impairs CNS angiogenesis and that EC-specific postnatal loss of Reck, combined with loss of Norrin, impairs blood-brain barrier (BBB) maintenance. The most N-terminal domain of Reck binds to the leucine-rich repeat (LRR) and immunoglobulin (Ig) domains of Gpr124, and weakening this interaction by targeted mutagenesis reduces Reck/Gpr124 stimulation of Wnt7a signaling in cell culture and impairs CNS angiogenesis. Finally, a soluble Gpr124(LRR-Ig) probe binds to cells expressing Frizzled, Wnt7a or Wnt7b, and Reck, and a soluble Reck(CC1-5) probe binds to cells expressing Frizzled, Wnt7a or Wnt7b, and Gpr124. These experiments indicate that Reck and Gpr124 are part of the cell surface protein complex that transduces Wnt7a- and Wnt7b-specific signals in mammalian CNS ECs to promote angiogenesis and regulate the BBB.
KW - CNS angiogenesis
KW - Wnt reporter
KW - blood-brain barrier
KW - canonical Wnt signaling
KW - mouse genetics
KW - vascular endothelial cells
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U2 - 10.1016/j.neuron.2017.07.031
DO - 10.1016/j.neuron.2017.07.031
M3 - Article
C2 - 28803732
AN - SCOPUS:85028349879
SN - 0896-6273
VL - 95
SP - 1056-1073.e5
JO - Neuron
JF - Neuron
IS - 5
ER -