Growth-associated protein 43 immunoreactivity in the superficial dorsal horn of the rat spinal cord is localized in atrophic C-fiber, and not in sprouted A-fiber, central terminals after peripheral nerve injury.
Peripheral nerve injury induces the up-regulation in dorsal root ganglion cells of growth-associated protein 43 (GAP-43) and its transport to the superficial laminae of the dorsal horn of the spinal cord, where it is located primarily in unmyelinated axons and growth-cone like structures. Peripheral nerve injury also induces the central terminals of axotomized myelinated axons to sprout and form novel synaptic contacts in lamina II of the dorsal horn. To investigate whether the sprouting of A-fiber central terminals into lamina II is the consequence of GAP-43 incorporation into their terminal membranes, we have used an ultrastructural analysis with double labelling to identify the localization of GAP-43 immunoreactivity. Transganglionic transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) was used to identify C-fiber terminals. Transganglionic transport of the B fragment of cholera toxin conjugated to horseradish peroxidase (B-HRP) was used to label A-fiber sciatic nerve central terminals in combination with GAP-43 immunocytochemistry. GAP-43 was found to colocalize only with WGA-HRP- and not with B-HRP-labelled synapses or axons. In addition, many single-labelled GAP-43 synapses were observed. Many of the WGA-HRP-labelled terminals that were characterized by degenerative changes were GAP-43 immunoreactive. Our results indicate that peripheral nerve injury induces novel synapse formation of A fibers in lamina II but that up-regulated levels of GAP-43 are present mainly in other axon projections to the superficial dorsal horn.
Doubell
T P
TP
Department of Anatomy and Developmental Biology, University College London, United Kingdom.
Woolf
C J
CJ
D000818:Animals; D001284:Atrophy; D001370:Axonal Transport; D002772:Cholera Toxin; D019922:GAP-43 Protein / Q000032:analysis / Q000096:biosynthesis*; D006735:Horseradish Peroxidase; D008297:Male; D016253:Microscopy, Immunoelectron; D009412:Nerve Fibers / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D009413:Nerve Fibers, Myelinated / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D009416:Nerve Regeneration; D051381:Rats; D017207:Rats, Sprague-Dawley; D012584:Sciatic Nerve / Q000293:injuries* / Q000502:physiology; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate
D019922:GAP-43 Protein; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate; C052635:cholera toxin, B subunit-horseradish peroxidase; D002772:Cholera Toxin; D006735:Horseradish Peroxidase