Changes in cytoskeletal gene expression affect the composition of regenerating axonal sprouts elaborated by dorsal root ganglion neurons in vivo.
The effect of a change in neurofilament (NF) and tubulin gene expression on the elongation of axonal sprouts by adult rat sensory neurons was examined. Distal sciatic nerve crush axotomy was used to initiate changes in cytoskeletal gene expression in lumbar dorsal root ganglion (DRG) neurons. In situ hybridization of DRG neurons with 35S-labeled cDNA probes revealed a significant reduction in the level of mRNAs for the low-molecular weight-NF protein and a significant increase in the level of beta tubulin mRNAs by 2 weeks after axotomy. A novel modification of the axonal transport paradigm was used to examine the biochemical composition of the regenerating axons formed by primed and unprimed DRG neurons. Primed neurons (which had sustained a crush axotomy of the distal sciatic nerve 2 weeks earlier) and unprimed (normal) neurons were labeled by microinjection of 35S-methionine and then stimulated to regenerate axons by a crush located very close to the DRG. In this paradigm, axonal sprouts that formed after the proximal crush axotomy incorporated radiolabeled, slow axonally transported proteins as they elongated. Fluorographs of SDS-PAGE revealed that the regenerating axonal sprouts of primed DRG cells incorporated and conveyed significantly less labeled NF protein than did the regenerating axons of unprimed DRG neurons. Electron microscopy revealed that the regenerating axonal sprouts of primed DRG cells contained numerous microtubules but very few identifiable NFs compared with the regenerating sprouts of unprimed DRG neurons.(ABSTRACT TRUNCATED AT 400 WORDS)
Oblinger
M M
MM
Department of Cell Biology and Anatomy, Chicago Medical School, Illinois 60064.
Szumlas
R A
RA
Wong
J
J
Liuzzi
F J
FJ
D000818:Animals; D001370:Axonal Transport; D001369:Axons / Q000378:metabolism / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology*; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D005786:Gene Expression Regulation; D007382:Intermediate Filaments / Q000502:physiology; D008297:Male; D008854:Microscopy, Electron; D009416:Nerve Regeneration; D009474:Neurons / Q000502:physiology / Q000648:ultrastructure; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D014404:Tubulin / Q000235:genetics
D012333:RNA, Messenger; D014404:Tubulin