The ability of axons to regenerate their growth cones depends on axonal type and age, and is regulated by calcium, cAMP and ERK.
The processes activated at the time of axotomy and leading to the formation of a new growth cone are the first step in regeneration, but are still poorly characterized. We investigated this event in an in vitro model of axotomy performed on dorsal root ganglia and retinal explants. We observed that the dorsal root ganglion axons and retinal ganglion cell axons, which had grown out on a poly d-lysine/laminin substrate at the time of culture preparation greatly differed in their regenerative response after a subsequent in vitro lesion made far from the cell body. The majority of axons of adult dorsal root ganglia but only a small percentage of axons of adult retinal ganglion cells regenerated new growth cones within four hours after in vitro axotomy, though both kinds of axons were growing before the lesion. The depletion of extracellular calcium and the inhibition of extracellular-signal regulated kinase 1,2 (ERK) and protein kinase A (PKA) at the time of injury significantly impaired the capacity of dorsal root ganglia axons to re-initiate growth cones without affecting growth cone motility. Pharmacological treatments directed at increasing the level of cAMP promoted growth cone regeneration in adult retinal ganglion cell axons in spite of the low regenerative potential exhibited in normal conditions. Understanding the cellular mechanisms activated at the time of lesion and leading to the formation of a new growth cone is necessary for devising treatments aimed at enhancing the regenerative response of injured axons.
Chierzi
Sabrina
S
Cambridge University Centre for Brain Repair, Robinson Way, Cambridge CB2 2PY, UK. sabrina.chierzi@in.cnr.it
Ratto
Gian Michele
GM
Verma
Poonam
P
Fawcett
James W
JW
D000199:Actins / Q000378:metabolism; D000375:Aging / Q000502:physiology*; D000818:Animals; D001369:Axons / Q000145:classification* / Q000502:physiology; D019771:Axotomy / Q000379:methods; D002070:Butadienes / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005260:Female; D019922:GAP-43 Protein / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D020439:Growth Cones / Q000187:drug effects / Q000502:physiology*; D007150:Immunohistochemistry / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D009416:Nerve Regeneration / Q000187:drug effects / Q000502:physiology*; D009570:Nitriles / Q000494:pharmacology; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000166:cytology / Q000187:drug effects; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000502:physiology; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
D000199:Actins; D002070:Butadienes; D004791:Enzyme Inhibitors; D019922:GAP-43 Protein; D009570:Nitriles; D014404:Tubulin; C113580:U 0126; D000242:Cyclic AMP; D048049:Extracellular Signal-Regulated MAP Kinases; D002118:Calcium