Axonal transport and metabolism of [ 3H]fucose- and [ 35S]- Sulfate-labeled macromolecules in the rat visual system

Abstract

The axonal transport of labeled macromolecules in retinal ganglion cells of rats was investigated from 1 to 20 days following intraocular injection of [ 3H]fucose and [ 35S]sulfate. Maximal incorporation of [ 3H]fucose into acid insoluble material in the retina was at 8 h, followed by a biphasic decline. Transported [ 3H]fucose (98% as glycoprotein) was in the optic nerve at 1 h, the optic tract and lateral geniculate body by 2 h, and the superior colliculus by 3 h after injection, indicating a rate of transport of approximately 200 mm/day. Radioactivity continued to accumulate in the superior colliculus for at least 8 h and began to decline rapidly by 24 h. Between 3 and 6 days levels rose again in both optic tract and superior colliculus before starting a gradual decline, indicating that a wave of rapidly transported material was delayed in leaving the retina. When proteins in the superior colliculus were fractionated by gel electrophoresis, the composition of the two fucosylated protein transport phases could be partially resolved. Radioactivity in individual gel peaks represented primarily in the first phase decayed with an average half-life of one day, although that in one prominent protein of molecular weight 280,000 turned over with a half-life of the order of 12 h. Radioactive peaks primarily in the second phase decayed with an average half-life of more than a week. Incorporation of [ 35S]sulfate into acid insoluble material in the retina was maximal at 1–2 h, after which there was a rapid loss of label. The appearance of [ 35S]sulfate in the optic tract, loateral geniculate body and superior colliculus preceded by a short time that of the [ 3H]fucose; indicating a shorter retinal processing time for this label. The total transported [ 35S]sulfate in the superior colliculus peaked by 4–8 h and had fallen by 65% at one day; no prominent second wave of transport was observed as was the case for [ 3H]fucose. Acid insoluble [ 35S]sulfate in the superior colliculus was equally divided between glycopeptides and glycosaminoglycans at all times examined, indicating that these macromolecules are transported at the same rate. [ 35S]Sulfate incorporated into various proteins fractionated by gel electrophoresis had heterogeneous turnover rates, the average being around 12 h. Radioactivity in one group of proteins, of molecular weight around 90,000, decayed with a half-life of only a few hours.