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The Journal of Neuroscience, April 15, 2002, 22(8):3144-3160
The Critical Role of Basement Membrane-Independent Laminin
1 Chain during Axon Regeneration in the CNS
Barbara Grimpe1, Sucai Dong3, Catherine Doller1, Katherine Temple2, Alfred T. Malouf2, and Jerry Silver1 Departments of 1 Neurosciences and 2 Pediatrics, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106, and 3 Department of Neurobiology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261
We have addressed the question of whether a family of axon growth-promoting molecules known as the laminins may play a role during axon regeneration in the CNS. A narrow sickle-shaped region containing a basal lamina-independent form of laminin exists in and around the cell bodies and proximal portion of the apical dendrites of CA3 pyramidal neurons of the postnatal hippocampus. To understand the possible function of laminin in axon regeneration within this pathway, we have manipulated laminin synthesis at the mRNA level in a slice culture model of the lesioned mossy system. In this model early postnatal mossy fibers severed near the hilus can regenerate across the lesion and elongate rapidly within strata lucidum and pyramidale. In slice cultures of the postnatal day 4 hippocampus, 2 d before lesion and then continuing for 1-5 d after lesion, translation of the
1 chain product of laminin was reduced by using antisense oligodeoxyribonucleotides and DNA enzymes. In the setting of the lesioned organotypic hippocampal slice, astroglial repair of the lesion and overall glial patterning were unperturbed by the antisense or DNA enzyme treatments. However, unlike controls, in the treated, lesioned slices the vast majority of regenerating mossy fibers could not cross the lesion site; those that did were very much shorter than usual, and they took a meandering course. In a recovery experiment in which the DNA enzyme or antisense oligos were washed away, laminin immunoreactivity returned and mossy fiber regeneration resumed. These results demonstrate the critical role of laminin(s) in an axon regeneration model of the CNS.
Key words: extracellular matrix; hippocampus; organotypic slice cultures; reactive astrocytes; glial scar; dendritic spines; axon guidance; antisense ODN; DNA enzyme
Copyright © 2002 Society for Neuroscience 0270-6474/02/2283144-17$05.00/0
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