Semaphorins and their receptors, plexins, have emerged as important regulators of a multitude of biological processes. Plexin-B3 has been shown to be selectively expressed in postnatal oligodendrocytes. In contrast to the well-characterized Plexin-A family and the Plexin-B family members Plexin-B1 and -B2, no data are available on the functional role of Plexin-B3 in the central nervous system in vivo. Here we have elucidated the functional significance of Plexin-B3 by generating and analyzing constitutive knock-out mice. Plexin-B3-deficient mice were found to be viable and fertile. A systematic histological analysis revealed no morphological defects in the brain or spinal cord of mutant animals. In detailed behavioural analyses of locomotor activity, motor coordination, motor learning, and anxiety levels Plexin-B3-deficient mice were indistinguishable from wild-type controls. Thus we conclude that under physiological conditions Plexin-B3 is not essential for the development and function of the central nervous system.