Ectopic adenoviral vector-directed expression of Sema3A in organotypic spinal cord explants inhibits growth of primary sensory afferents.
Sema3A (Sema III, SemD, collapsin-1) can induce neuronal growth cone collapse and axon repulsion of distinct neuronal populations. To study Sema3A function in patterning afferent projections into the developing spinal cord, we employed the recombinant adenoviral vector technique in embryonic rat spinal cord slices. Virus solution was injected in the dorsal aspect of organotypic spinal cord cultures with segmentally attached dorsal root ganglia (sc-DRG). In cultures grown in the presence of nerve growth factor (NGF), injected either with the control virus AdCMVLacZ or with vehicle only, afferent innervation patterns were similar to those of control. However, unilateral injection of AdCMVSema3A/AdCMVLacZ in sc-DRG slices revealed a strong inhibitory effect on NGF-dependent sensory afferent growth. Ectopic Sema3A in the dorsal spinal cord, the target area of NGF-responsive DRG fibers in vivo, created an exclusion zone for these fibers and as a result they failed to reach and innervate their appropriate target zones. Taken together, gain of Sema3A function in the dorsal aspect of sc-DRG cultures revealed a dominant inhibitory effect on NGF-dependent, nociceptive sensory DRG afferents, an observation in line with the model proposed by E. K. Messersmith et al. (1995, Neuron 14, 949-959), suggesting that Sema3A secreted by spinal cord cells can act to repel central sensory fibers during the formation of lamina-specific connections in the spinal cord.
Pasterkamp
R J
RJ
Graduate School Neurosciences Amsterdam, The Netherlands Institute for Brain Research, Amsterdam, 1105 AZ, The Netherlands.
Giger
R J
RJ
Baker
R E
RE
Hermens
W T
WT
Verhaagen
J
J
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015152:Blotting, Northern; D019556:COS Cells; D002633:Chemotaxis; D002828:Choristoma; D046508:Culture Techniques; D005727:Ganglia, Spinal / Q000196:embryology* / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D020932:Nerve Growth Factor / Q000378:metabolism; D009475:Neurons, Afferent / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D040121:Semaphorin-3A; D013116:Spinal Cord / Q000196:embryology* / Q000378:metabolism
D006023:Glycoproteins; D040121:Semaphorin-3A; D020932:Nerve Growth Factor