Axo-glial interactions at the dorsal root transitional zone regulate neurofilament protein synthesis in axotomized...

After dorsal root crush, dramatic ultrastructural differences are observed between regenerated dorsal root axonal endings that are physically blocked at a ligation neuroma and those that are allowed to form axo-glial endings among the astrocytes at the dorsal root transitional zone (DRTZ). Physically blocked axonal endings swell immensely with membranous organelles and neurofilaments (NFs) while axo-glial endings do not, suggesting that DRTZ astrocytes stop axonal growth by activating a physiological stop pathway within those endings. Since protease-dependent NF degradation at axonal endings is a part of this pathway, this study addresses the question of whether NF subunit synthesis in the dorsal root ganglion (DRG) is regulated by the pathway. Lumbar dorsal roots were crushed and, at various postinjury times, the attached DRGs were removed and pulse-labeled in vitro with 35S-methionine for subsequent analysis of protein synthesis by electrophoresis and fluorography. Within 24 hr of axotomy, there was a down-regulation of the 68 kDa (NF-L) and 145 kDa (NF-M) NF subunits. At 14 d postcrush, a time when most of the regenerating axons have reached and been stopped by DRTZ astrocytes, NF protein synthesis returned to control levels. By contrast, when the axons were prevented from reaching the DRTZ by ligating or removing segments of the roots, NF synthesis failed to return to normal levels. These data suggest that activation of the physiological stop pathway by DRTZ astrocytes regulates NF protein synthesis in the DRG.