The downregulation of GAP-43 is not responsible for the failure of regeneration in freeze-killed nerve grafts in the rat.
Freeze-killed nerve grafts in rats are able to support limited axonal regeneration from severed peripheral nerves, but by 6 weeks postoperation, axonal elongation through the grafts ceases. To find out whether this limited regeneration may be related to GAP-43 expression, 4-cm freeze-killed nerve grafts were attached to the proximal stumps of severed tibial nerves in adult inbred Fischer rats. For comparison, tibial nerve crush, to allow functional regeneration, or section and ligation, which allows only abortive axonal sprouting, were also performed. After survival for 3 or 6 weeks, the lumbar spinal cord and L4 dorsal root ganglia were stained for GAP-43 mRNA. Freeze-killed grafts of 3-8 weeks duration were processed for GAP-43 immunocytochemistry. Three weeks after all three operations, comparable numbers of axotomized spinal motorneurons and primary sensory DRG neurons reexpressed high levels of GAP-43 mRNA. Six weeks after tibial nerve crush, the number of tibial motorneurons and DRG cells expressing GAP-43 mRNA returned to control levels but after section and ligation or freeze-killed nerve grafting many positively stained cells were still visible. GAP-43 immunoreactivity was detectable using immunocytochemistry in many unmyelinated axons which had regenerated into the freeze-killed grafts at all times. Both axonal profiles in contact with Schwann cells and those which lacked such contact were GAP-43 positive. These results suggest that the cessation of axonal regeneration into freeze-killed tibial nerve grafts is not the result of a down-regulation of GAP-43. Furthermore, the presence of high levels of GAP-43 alone is not sufficient to ensure prolonged axonal regeneration.
Department of Anatomy and Developmental Biology, University College London, England.
M S
MS
Chong
C J
CJ
Woolf
P
P
Andrews
M
M
Turmaine
D J
DJ
Schreyer
P N
PN
Anderson
D000818:Animals; D001369:Axons / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D005612:Freeze Drying; D019922:GAP-43 Protein; D005727:Ganglia, Spinal / Q000378:metabolism / Q000502:physiology; D015870:Gene Expression; D017403:In Situ Hybridization; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis / Q000096:biosynthesis*; D008854:Microscopy, Electron; D009409:Nerve Crush; D009416:Nerve Regeneration; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000096:biosynthesis*; D016900:Neurofilament Proteins / Q000096:biosynthesis; D010525:Peripheral Nerves / Q000378:metabolism / Q000637:transplantation*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D011916:Rats, Inbred F344; D013116:Spinal Cord / Q000378:metabolism / Q000502:physiology; D013979:Tibial Nerve / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation*; D013997:Time Factors
D019922:GAP-43 Protein; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D012333:RNA, Messenger