L1, beta1 integrin, and cadherins mediate axonal regeneration in the embryonic spinal cord.
Embryonic birds and mammals are capable of axon regeneration after spinal cord injury, but this ability is lost during a discrete developmental transition. We recently showed that changes within maturing neurons, as opposed to changes solely in the spinal cord environment, significantly restrict axon regeneration during development. The developmental changes within neurons that limit axon regeneration remain unclear. One gap in knowledge is the identity of the adhesive receptors that embryonic neurons use to extend axons in the spinal cord. Here we test the roles of L1/NgCAM, beta1 integrin, and cadherins, using a coculture system in which embryonic chick brainstem neurons regenerate axons into an explant of embryonic spinal cord. By in vivo and in vitro methods, we found that brainstem neurons reduce axonal expression of L1 as they mature. Disrupting either L1 or beta1 integrin function individually in our coculture system partially inhibited growth of brainstem axons in spinal cords, while disrupting cadherin function alone had no effect. However, when all three adhesive receptors were blocked simultaneously, axon growth in the spinal cord was reduced by 90%. Using immunohistochemistry and in situ hybridization we show that during the period when neurons lose their regenerative capacity they reduce expression of mRNA for N-cadherin, and reduce axonal L1/NgCAM protein through a post-transcriptional mechanism. These data show that embryonic neurons use L1/NgCAM, beta1 integrin, and cadherin receptors for axon regeneration in the embryonic spinal cord, and raise the possibility that a reduced expression of these essential receptors may contribute to the low-regenerative capacity of older neurons.
Blackmore
Murray
M
Department of Neuroscience, University of Minnesota, Minneapolis, Minnesota 55455, USA. mblackmore@med.miami.edu
Letourneau
Paul C
PC
D000367:Age Factors; D000596:Amino Acids; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001369:Axons / Q000187:drug effects / Q000502:physiology*; D001933:Brain Stem; D015820:Cadherins / Q000276:immunology / Q000502:physiology*; D002118:Calcium / Q000494:pharmacology; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry / Q000379:methods; D019012:Integrin beta1 / Q000276:immunology / Q000502:physiology*; D039842:Neural Cell Adhesion Molecule L1 / Q000276:immunology / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology; D009924:Organ Culture Techniques; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D013116:Spinal Cord / Q000166:cytology* / Q000196:embryology; D014162:Transfection / Q000379:methods
D000596:Amino Acids; D000906:Antibodies; D015820:Cadherins; D019012:Integrin beta1; D039842:Neural Cell Adhesion Molecule L1; C098463:dolaisoleucine; D002118:Calcium