EphB receptor tyrosine kinases control multiple steps in nervous system development. However, it remains unclear whether EphBs regulate these different developmental processes directly or indirectly. In addition, given that EphBs signal through multiple mechanisms, it has been challenging to define which signaling functions of EphBs regulate particular developmental events. To address these issues, we engineered triple knock-in mice in which the kinase activity of three neuronally expressed EphBs can be rapidly, reversibly and specifically blocked. We found that the tyrosine kinase activity of EphBs was required for axon guidance in vivo. In contrast, EphB-mediated synaptogenesis occurred normally when the kinase activity of EphBs was inhibited, suggesting that EphBs mediate synapse development by an EphB tyrosine kinase–independent mechanism. Taken together, our data indicate that EphBs control axon guidance and synaptogenesis by distinct mechanisms and provide a new mouse model for dissecting EphB function in development and disease.
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We thank M. Thompson, Y. Zhou and H. Ye of the Children's Hospital Boston Intellectual and Developmental Disabilities Research Center Mouse Gene Manipulation Core for ES cell work and blastocyst injection, T. Kuwajima and members of C. Mason's laboratory for advice on retinal explants and DiI labeling, M. Lopez for help selecting and generating PP1 analogs, Z. Wills and A. Mardinly for help with synapse analysis, S. Cohen for advice on electrophysiological recordings, P. Zhang for assistance with animal management, and L. Hu for antibody work. This research was funded by US National Institutes of Health grants RO1-NS-045500 (M.E.G.) and RO1-EY-018207 (C.W.C.). H.-Y.H.H. was supported by the Marion Abbe Fellowship of the Damon Runyon Cancer Research Foundation and US National Institutes of Health training grants in neurodegeneration and cancer biology. M.J.S. was supported by a National Science Foundation Graduate Research Fellowship. M.A.R. was supported by a training grant from the National Institute on Drug Abuse (T32 DA07290).
The authors declare no competing financial interests.
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Soskis, M., Ho, H., Bloodgood, B. et al. A chemical genetic approach reveals distinct EphB signaling mechanisms during brain development. Nat Neurosci 15, 1645–1654 (2012) doi:10.1038/nn.3249
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