Intracellular transport and neuronal activation of phospholipid and glycoprotein synthesis during axonal regeneration of cranio-spinal nerves

In the present work, the hypothesis that the increased rapid intracellular transport of newly-synthesized material along the axons of a regenerating system is sustained by an alteration of the transport of proteolipid complexes through subcellular compartments of a neuronal cell body was tested by a biochemical methodology. The motoneurons of spinal cord ventral horn, 4 wk after unilateral lesion (crush) of cervico-thoracic nerves of the rabbit at the level of brachial plexus, were chosen as the model system of regeneration. A time-staggered procedure of in vivo and in vitro double labeling with metabolic precursors, such as [³H]-choline, [¹⁴C]-choline, [³H]-fucose, and [¹⁴C]-fucose, was used. Subcellular fractions (RER, SER, Golgi apparatus, and plasma membranes) of ventral horn tissue, taken from spinal cord hemisections (regenerating and contralateral side), were further isolated. Twenty-eight days after axotomy, we did not observe any change of intracellular transport kinetics (¹⁴C/³H ratio) of newly-synthesized choline-phospholipids and glycoproteins in regenerating motoneurons compared to controls. However, associated with regenerating phenomenon in Golgi apparatus, we observed an increase of labeled choline-phospholipid and glycoprotein material that could contribute to the increased fast axonal transport and delivery of membrane proteolipid complexes to plasma membrane and axonal compartments. The increase of glycoprotein labeling was more pronounced in the SER portion (vesicles and elements of smooth membranes). This result is in favor of the hypothesis that membrane-bound proteins are transported from the Golgi to the axon through the perikaryal SER.