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The Journal of Neuroscience, September 3, 2003, 23(22):8176-8184
 Previous Article
Slit2 Guides Both Precrossing and Postcrossing Callosal Axons at the Midline In Vivo
Tianzhi Shu,1
Vasi Sundaresan,4
Margaret M. McCarthy,2,3 and
Linda J. Richards1,3
Departments of 1Anatomy and Neurobiology and
2Physiology and 3Program in
Neuroscience, The University of Maryland, Baltimore, School of Medicine,
Baltimore, Maryland 21201, and 4Department of
Histopathology, St. Thomas' Hospital, London SE1 7EH, United Kingdom
Commissural axons generally cross the midline only once. In the
Drosophila nerve cord and mouse spinal cord, commissural axons are
guided by Slit only after they cross the midline, where Slit prevents these
axons from recrossing the midline. In the developing corpus callosum, Slit2
expressed by the glial wedge guides callosal axons before they cross the
midline, as they approach the corticoseptal boundary. These data highlighted a
potential difference between the role of Slit2 in guiding commissural axons in
the brain compared with the spinal cord. Here, we investigate whether Slit2
also guides callosal axons after they cross the midline. Because such
questions cannot be addressed in conventional gene knock-out animals, we used
in utero injections of antisense oligonucleotides to specifically
deplete Slit2 on only one side of the brain. We used this technique together
with a novel in vitro assay of hemisected brain slices to
specifically analyze postcrossing callosal axons. We find that in the brain,
unlike the spinal cord, Slit2 mediates both precrossing and postcrossing
axonal guidance. Depletion of Slit2 on one side of the brain causes axons to
defasciculate and, in some cases, to aberrantly enter the septum. Because
these axons do not recross the midline, we conclude that the principle
function of Slit2 at the cortical midline may be to channel the axons along
the correct path and possibly repel them away from the midline. We find no
evidence that Slit2 prevents axons from recrossing the midline in the
brain.
Key words: corpus callosum; commissure; axon guidance; cortical development; glial wedge; midline; Slit; Robo; antisense oligonucleotides
Received March 12, 2003;
revised July 18, 2003;
accepted July 21, 2003.
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