Skip to main content
Log in

Expression of GAP-43 mRNA in the adult mammalian spinal cord under normal conditions and after different types of lesions, with special reference to motoneurons

  • Published:
Experimental Brain Research Aims and scope Submit manuscript

Summary

In situ hybridization histochemistry was used to detect cell bodies expressing mRNA encoding for the phosphoprotein GAP-43 in the lumbosacral spinal cord of the adult rat, cat and monkey under normal conditions and, in the cat and rat, also after different types of lesions. In the normal spinal cord, a large number of neurons throughout the spinal cord gray matter were found to express GAP-43 mRNA. All neurons, both large and small, in the motor nucleus (Rexed's lamina IX) appeared labeled, indicating that both alpha and gamma motoneurons express GAP-43 mRNA under normal conditions. After axotomy by an incision in the ventral funiculus or a transection of ventral roots or peripheral nerves, GAP-43 mRNA was clearly upregulated in axotomized motoneurons, including both alpha and gamma motoneurons. An increase in GAP-43 mRNA expression was already detectable 24 h postoperatively in lumbar motoneurons both after a transection of the sciatic nerve at knee level and after a transection of ventral roots. At this time, a stronger response was seen in the motoneurons which had been subjected to the distal sciatic nerve transection than was apparent for the more proximal ventral root lesion. An upregulation of GAP-43 mRNA could also be found in intact motoneurons located on the side contralateral to the lesion, but only after a peripheral nerve transection, indicating that the concomitant influence of dorsal root afferents may play a role in GAP-43 mRNA regulation. However, a dorsal root transection alone did not seem to have any detectable influence on the expression of GAP-43 mRNA in spinal motoneurons, while the neurons located in the superficial laminae of the dorsal horn responded with an upregulation of GAP-43 mRNA. The presence of high levels of GAP-43 in neurons has been correlated with periods of axonal growth during both development and regeneration. The role for GAP-43 in neurons under normal conditions is not clear, but it may be linked with events underlying remodelling of synaptic relationships or transmitter release. Our findings provide an anatomical substrate to support such a hypothesis in the normal spinal cord, and indicate a potential role for GAP-43 in axon regeneration of the motoneurons, since GAP-43 mRNA levels was strongly upregulated following both peripheral axotomy and axotomy within the spinal cord. The upregulation of GAP-43 mRNA found in contralateral, presumably uninjured motoneurons after peripheral nerve transection, as well as in dorsal horn neurons after a dorsal root transection, indicates that GAP-43 levels are altered not only as a direct consequence of a lesion, but also after changes in the synaptic input to the neurons.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Akers RF, Routtenberg A (1987) Calcium-promoted translocation of protein kinase C to synaptic membranes: relation to the phosphorylation of an endogenous substrate (protein F1) involved in synaptic plasticity. J Neurosci 7:3976–3983

    Google Scholar 

  • Aloyo VJ, Zwiers H, Gispen WH (1983) Phosphorylation of B-50 protein by calcium-activated, phopholipid-dependent protein kinase and B-50 protein kinase. J Neurochem 41:649–653

    Google Scholar 

  • Arvidsson U, Risling M, Cullheim S, Dagerlind Å, Lindå H, Shupliakov O, Ulfhake B, Hökfelt T (1992) On the distribution of GAP-43 and its relation to serotonin in the adult monkey and cat spinal cord and lower brain stem. Eur J Neurosci (in press)

  • Bendotti C, Servadio A, Samanin R (1991) Distribution of GAP-43 mRNA in the brain stem of adult rats as evidenced by in situ hybridization: localization within monoaminergic neurons. J Neurosci 11:600–607

    Google Scholar 

  • Benowitz LI, Apostolides PJ, Perrone-Bizzozero N, Finklestein SP, Zwiers H (1988) Anatomical distribution of the growth associated protein GAP-43/B-50 in the adult rat brain. J Neurosci 8:339–352

    Google Scholar 

  • Benowitz LI, Perrone-Bizzozero NI (1991) The relationship of GAP-43 to the development and plasticity of synaptic connections. Ann N Y Acad Sci 627:58–74

    Google Scholar 

  • Benowitz LI, Routtenberg A (1987) A membrane phosphoprotein associated with neuronal development, axonal regeneration, phopholipid metabolism and synaptic plasticity. Trends Neurosci 10:527–532

    Google Scholar 

  • Bisby MA (1988) Dependence of GAP43 (B50, F1) transport on axonal regeneration in rat dorsal root ganglion neurons. Brain Res 458:157–161

    Google Scholar 

  • Carlstedt T, Lindå H, Cullheim S, Risling M (1986) Reinnervation of hindlimb muscles after ventral root avulsion and implantation in the lumbar spinal cord of the adult rat. Acta Physiol Scand 128:645–646

    CAS  PubMed  Google Scholar 

  • Cullheim S, Carlstedt T, Lindå H, Risling M, Ulfhake B (1989) Motoneurons reinnervate skeletal muscle after ventral root implantation into the spinal cord of the cat. Neuroscience 29:725–733

    Google Scholar 

  • Cunha A da, Aloya VJ, Vitkovic L (1991) Developmental regulation of GAP-43, glutamine synthetase and beta-actin mRNA in rat cortical astrocytes. Dev Brain Res 64:212–215

    Google Scholar 

  • Cunha A da, Vitkovic L (1990) Regulation of immunoreactive GAP-43 expression in rat cortical macroglia is cell type specific. J Cell Biol 111:209–215

    Google Scholar 

  • Curtis R, Hardy R, Reynolds R, Spruce BA, Wilkin GP (1991) Down-regulation of GAP-43 during oligodendrocyte development and lack of expression by astrocytes in vivo: implications for macroglial differentiation. Eur J Neurosci 3:876–886

    Google Scholar 

  • Dagerlind Å, Friberg K, Bean AJ, Hökfelt T (1992) Sensitive mRNA detection using unfixed tissue: combined radioactive and non-radioactive in situ hybridization histochemistry. Histochemistry (in press)

  • Dagerlind Å, Schalling M, Eneroth P, Goldstein M, Hökfelt T (1990) Effects of reserpine on phenylethanolamine N-methyltransferase mRNA levels in rat adrenal gland: role of steroids. Neurochem Int 17:343–356

    Google Scholar 

  • Dekker LV, De Graan PNE, Versteeg DHG, Oestreicher AB, Gispen WH (1989) Inhibition of noradrenaline release by antibodies to B-50 (GAP-43). Nature 342:74–76

    Google Scholar 

  • Deloulme J-C, Janet T, Au D, Storm DR, Sensenbrenner M (1990) Neuromodulin (GAP-43): a neuronal protein kinase C substrate is also present in O-2 A glial cell lineage. Characterization of neuromodulin in secondary cultures of oligodendrocytes and comparison with the neuronal antigen. J Cell Biol 111:1559–1569

    Google Scholar 

  • Doster SK, Lozano AM, Aguayo AJ, Willard MB (1991) Expression of the growth associated protein GAP-43 in adult rat retinal ganglion cells following axon injury. Neuron 6:635–647

    Google Scholar 

  • Ernfors P, Henschen A, Olson L, Persson H (1989) Expression of nerve growth factor receptor mRNA is developmentally regulated and increased after axotomy in rat spinal cord motoneurons. Neuron 2:1605–1613

    Google Scholar 

  • Jacobsson RD, Virag I, Skene JHP (1986) A protein associated with axon growth, GAP-43, is widely distributed and developmentally regulated in rat CNS. J Neurosci 6:1843–1855

    Google Scholar 

  • Kalil K, Skene JHP (1986) Elevated synthesis of an axonally transported protein correlates with axon outgrowth in normal and injured pyramidal tracts. J Neurosci 6:2563–2570

    Google Scholar 

  • Karns LR, Ng S-C, Freeman JA, Fishman MC (1987) Cloning of complementary DNA for GAP-43, a neuronal growth-related protein. Science 236:597–600

    Google Scholar 

  • Koliatsos VE, Crawford TO, Price DL (1991) Axotomy induces nerve growth factor receptor immunoreactivity in spinal motor neurons. Brain Res 549:297–304

    Google Scholar 

  • Lindå H, Risling M, Cullheim S (1985) “Dendraxons” in regenerating motoneurons in the cat: do dendrites generate new axons after central axotomy. Brain Res 358:329–333

    Google Scholar 

  • Lovinger DM, Akers RF, Nelson RB, Barnes CA, McNaughton BL, Routtenberg A (1985) A selective increase in phosphorylation of protein F1, a protein kinase C substrate, directly related to three day growth of long term synaptic enhancement. Brain Res 342:137–143

    Google Scholar 

  • Lozano AM, Doster SK, Aguayo AJ, Willard MB (1987) Immunoreactivity to GAP-43 in axotomized and regenerating retinal ganglion cells of adult rats. Soc Neurosci Abstr 13:1389

    Google Scholar 

  • Meberg PJ, Routtenberg A (1991) Selective expression of protein F1/(GAP-43) mRNA in pyramidal but not granule cells of the hippocampus. Neuroscience 45:721–733

    Google Scholar 

  • Meiri KF, Burkick D (1991) Nerve growth factor stimulation of GAP-43 phosphorylation in intact isolated growth cones. Neuroscience 11:3155–3164

    Google Scholar 

  • Oestreicher AB, Gispen WH (1986) Comparison of immunocytochemical distribution of the phophoprotein B-50 in the cerebellum and hippocampus of immature and adult rat brain. Brain Res 375:267–279

    Google Scholar 

  • Piehl F, Arvidsson U, Johnson H, Cullheim S, Villar M, Dagerlind Å, Terenius L, Hökfelt T, Ulfhake B (1991) Calcitonin generelated peptide (CGRP)-like immunoreactivity and CGRP mRNA in rat spinal cord motoneurons after different types of lesions. Eur J Neurosci 3:737–757

    Google Scholar 

  • Redshaw JD, Bisby MA (1984a) Fast axonal transport in central nervous system and peripheral nervous system axons following axotomy. J Neurobiol 15:109–117

    Google Scholar 

  • Redshaw JD, Bisby MA (1984b) Proteins of fast axonal transport in the regenerating hypoglossal nerve of the rat. Can J Physiol Pharmacol 62:1387–1393

    Google Scholar 

  • Reh TA, Redshaw JD, Bisby MA (1987) Axons of the pyramidal tract do not increase their transport of growth associated proteins after axotomy. Mol Brain Res 2:1–6

    Google Scholar 

  • Richardson PM, Issa VMK, Aguayo AJ (1982) Regeneration and retrograde degeneration of axons in the rat optic nerve. J Neurocytol 11:949–966

    Google Scholar 

  • Richardson PM, Issa VMK, Aguayo AJ (1984) Regeneration of long spinal axons in the rat. J Neurocytol 13:165–182

    Google Scholar 

  • Risling M, Cullheim S, Hildebrand C (1983) Reinnervation of the ventral root L7 from ventral horn neurons following intramedullary axotomy in adult cats. Brain Res 280:15–23

    Google Scholar 

  • Risling M, Fried K, Lindå H, Cullheim S, Meier M (1992) Changes in nerve growth factor receptor-like immunoreactivity in the spinal cord after ventral funiculus lesion in adult cats. J Neurocytol 21:79–93

    Google Scholar 

  • Risling M, Lindå H, Cullheim S, Franson P (1989) A persistent defect in the blood brain barrier after ventral funiculus lesion in adult cats implications for CNS regeneration. Brain Res 494:13–21

    Google Scholar 

  • Rosenthal A, Chan SY, Henzel W, Haskell C, Kuang WJ, Chen E, Wilcox JN, Ullrich A, Goeddel DV, Routtenberg A (1987) Primary structure and mRNA localization of protein F1, a growth-related protein kinase C substrate associated with synaptic plasticity. EMBO J 6:3641–3646

    Google Scholar 

  • Schalling M (1990) In situ hybridization studies on regulatory molecules in neural and endocrine tissues with special reference to expression of coexisting peptides. Thesis, Karolinska Institute, Stockholm

    Google Scholar 

  • Skene JHP (1989) Axonal growth-associated proteins. Annu Rev Neurosci 12:127–156

    Google Scholar 

  • Skene JHP, Willard M (1981) Axonally transported proteins associated with axon growth in rabbit central and peripheral nervous systems. J Cell Biol 89:96–103

    Google Scholar 

  • So KF, Aguayo AJ (1985) Lengthy regrowth of cut axons from ganglion cells after peripheral nerve transplantation into the retina of adult rats. Brain Res 328:349–354

    Google Scholar 

  • Sunderland S (1978) Nerves and nerve injuries. Churchill Livingstone, Edinburgh

    Google Scholar 

  • Takano K (1976) Absence of gamma-spindle loop in reinnervated hind leg muscles of the cat: “alpha-muscle”. Exp Brain Res 26:342–354

    Google Scholar 

  • Tetzlaff W, Alexander SW, Miller FD, Bisby MA (1991) Response of facial and rubrospinal neurons to axotomy: changes in mRNA expression for cytoskeleton proteins and GAP-43. J Neurosci 11:2528–44

    Google Scholar 

  • Tetzlaff W, Tsui BJ, Balfour JK (1990) Rubrospinal neurons increase GAP-43 and tubulin mRNA after cervical but not after thoracic axotomy. Soc Neurosci Abstr 16:338

    Google Scholar 

  • Tetzlaff W, Zwiers H, Lederis K, Cassar L, Bisby MA (1989) The axonal transport and localization of B50/GAP-43-like immunoreactivity in regenerating sciatic and facial nerves of the rat. J Neurosci 9:1303–1313

    Google Scholar 

  • Thoenen H (1991) The changing scene of neurotrophic factors. Trends Neurosci 14:165–170

    Article  CAS  PubMed  Google Scholar 

  • Thulin C-A (1960) Electrophysiological studies of peripheral nerve regeneration with special reference to the small diameter (gamma) fibers. Exp Neurol 2:598–612

    Google Scholar 

  • van der Zee CE, Nielander HB, Vos JP, Lopes da Silva S, Verhaagen J, Oestreicher AB, Schrama LH, Schotman P, Gispen WH (1989) Expression of growth-associated protein B-50 (GAP-43) in dorsal root ganglia and sciatic nerve during regenerative sprouting. J Neurosci 9:3505–12

    Google Scholar 

  • Verge VM, Tetzlaff W, Richardson PM, Bisby MA (1990) Correlation between GAP-43 and nerve growth factor receptors in rat sensory neurons. J Neurosci 10:926–934

    Google Scholar 

  • Verhaagen J, Oestreicher AB, Edwards PM, Veldman H, Jennekens FG, Gispen WH (1988) Light and electron microscopic study of phosphoprotein B-50 following denervation and reinnervation of the rat soleus muscle. J Neurosci 8:1759–1766

    Google Scholar 

  • Wood SJ, Pritchard J, Sofroniew MW (1990) Re-expression of nerve growth factor receptor after axonal injury recapitulates a developmental event in motor neurons: differential regulation when regeneration is allowed or prevented. Eur J Neurosci 2:650–657

    Google Scholar 

  • Yan Q, Snider WD, Pinzone JJ, Johnson EM (1988) Retrograde transport of nerve growth factor (NGF) in motoneurons of developing rats: assessment of potential neurotrophic effects. Neuron 1:335–343

    Google Scholar 

  • Yankner BA, Benowitz LI, Villa Komaroff L, Neve RL (1990) Transfection of PC12 cells with the human GAP-43 gene: effects on neurite outgrowth and regeneration. Mol Brain Res 7:39–44

    Google Scholar 

  • Young IWS (1990) In situ hybridization histochemistry. In: Björklund A, Hökfelt T, Wouterlood FG, Van der Pol AN (eds) Handbook of chemical neuroanatomy, vol 8. Analysis of neuronal microcircuits and synaptic interactions. Elsevier, Amsterdam, pp 481–512

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lindå, H., Piehl, F., Dagerlind, Å. et al. Expression of GAP-43 mRNA in the adult mammalian spinal cord under normal conditions and after different types of lesions, with special reference to motoneurons. Exp Brain Res 91, 284–295 (1992). https://doi.org/10.1007/BF00231661

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00231661

Key words

Navigation