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The Journal of Neuroscience, August 30, 2006, 26(35):8909-8914; doi:10.1523/JNEUROSCI.1569-06.2006

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Brief Communications
EphB Receptors and Ephrin-B3 Regulate Axon Guidance at the Ventral Midline of the Embryonic Mouse Spinal Cord

Stephanie R. Kadison,¹ Taija Mäkinen,³ Rüdiger Klein,³ Mark Henkemeyer,⁴ and Zaven Kaprielian^1,2

Departments of ¹Neuroscience and ²Pathology, Albert Einstein College of Medicine, Bronx, New York 10461, ³Department of Molecular Neurobiology, Max Planck Institute of Neurobiology, 82152 Martinsried, Germany, ⁴Center for Developmental Biology and Kent Waldrep Foundation Center for Basic Research on Nerve Growth and Regeneration, University of Texas Southwestern Medical Center, Dallas, Texas 33136

Correspondence should be addressed to Stephanie R. Kadison, Department of Neuroscience, Albert Einstein College of Medicine, Kennedy Center, Room 616, 1410 Pelham Parkway, Bronx, NY 10461. Email: skadison{at}umich.edu

EphB receptors and their ephrin-B ligands are required for midline guidance decisions at several rostrocaudal levels of the developing CNS. In the embryonic vertebrate spinal cord, ephrin-B3 is localized to the floor plate (FP) at the ventral midline (VM), ephrin-B1 and ephrin-B2 are expressed in the dorsal spinal cord, and decussated EphB receptor-bearing commissural axons navigate between these ventral and dorsal ephrin-B domains. Despite these compelling expression patterns, the in vivo role(s) for EphB and ephrin-B proteins in regulating the guidance of spinal commissural axons has not been established. Here, we use DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) labeling to assess the pathfinding of commissural axons in the spinal cords of ephrin-B and EphB mutant mouse embryos. In mice lacking ephrin-B3 or multiple EphB receptors, a significant number of axons followed aberrant trajectories in the immediate vicinity of the VM. Furthermore, forked transverse commissural (FTC) axons, a unique class of commissural axons that continues to project in the transverse plane on the contralateral side of the FP, were present at a markedly higher frequency in ephrin-B3 and EphB mutants, compared with wild-type embryos. Neither the midline guidance errors nor excessive numbers of FTC axons were observed in the spinal cords of ephrin-B3^lacz mice that express a truncated form of ephrin-B3, which is capable of forward but not reverse signaling. In contrast to the midline guidance defects observed in EphB and ephrin-B3 mutant embryos, wild-type-like contralateral projections were observed in mice lacking ephrin-B1 and/or ephrin-B2.

Key words: EphB; ephrin-B; mouse embryo; spinal cord; ventral midline; floor plate; commissural axon

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Received April 11, 2006; revised July 21, 2006; accepted July 25, 2006.

Correspondence should be addressed to Stephanie R. Kadison, Department of Neuroscience, Albert Einstein College of Medicine, Kennedy Center, Room 616, 1410 Pelham Parkway, Bronx, NY 10461. Email: skadison{at}umich.edu

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