The increase in B-50/GAP-43 in regenerating rat sciatic nerve occurs predominantly in unmyelinated axon shafts: a quantitative ultrastructural study.
The growth-associated protein B-50/GAP-43 is thought to play a crucial role in axonal growth. We investigated, by quantitative immunoelectron microscopy, whether there are differences in the subcellular distribution of B-50 in unmyelinated and myelinated axons of intact and regenerating sciatic nerves. Adult rats received an unilateral sciatic nerve crush and were euthanized 8 days later. Nerve pieces proximal from the crush site were embedded, and B-50 was visualized by specific B-50 antibodies and immunogold detection in ultrathin sections. The density of B-50 at the plasma membrane of unmyelinated axon shafts was significantly increased in the ipsilateral regenerating nerve in comparison to that of the contralateral intact nerve. In contrast, there was no significant difference in the B-50 density at the axolemma of myelinated regenerating and intact axon shafts. In the contralateral intact nerve, more B-50 was associated with the axolemma of unmyelinated axons than with the plasma membrane of myelinated axons. The density of axoplasmic B-50 was similar in intact unmyelinated and myelinated axon shafts, but was higher in regenerating nerve than in intact nerve. This suggests that enhanced axonal transport of B-50 occurs during axon outgrowth. Our study demonstrates a differential subcellular distribution of B-50 in unmyelinated and myelinated axon shafts in both the intact and regenerating sciatic nerve, indicating a differential inducible capacity for remodeling of the axon shafts.
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Rudolf Magnus Institute for Neurosciences, University of Utrecht, The Netherlands.
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D000818:Animals; D001369:Axons / Q000378:metabolism* / Q000648:ultrastructure; D007839:Functional Laterality / Q000502:physiology; D019922:GAP-43 Protein; D006133:Growth Substances / Q000378:metabolism*; D015151:Immunoblotting; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D009186:Myelin Sheath / Q000502:physiology; D009409:Nerve Crush; D009416:Nerve Regeneration / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012584:Sciatic Nerve / Q000502:physiology* / Q000648:ultrastructure; D013223:Statistics as Topic; D013347:Subcellular Fractions / Q000737:chemistry*
D019922:GAP-43 Protein; D006133:Growth Substances; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins