Evidence that all newly synthesized proteins destined for fast axonal transport pass through the Golgi apparatus.
Effects of the sodium ionophore, monensin, were examined on the passage from neuronal cell body to axon of materials undergoing fast intracellular transport. In vitro exposure of bullfrog dorsal root ganglia to concentrations of drug less than 1.0 micron led to a dose-dependent depression in the amount of fast-transported [3H]leucine- or [3H]glycerol-labeled material appearing in the nerve trunk. Incorporation of either precursor was unaffected. Exposure of a desheathed nerve trunk to similar concentrations of monensin, while ganglia were incubated in drug-free medium, had no effect on transport. With [3H]fucose as precursor, fast transport of labeled glycoproteins was depressed to the same extent as with [3H]leucine; synthesis, again, was unaffected. By contrast, with [3H]galactose as precursor, an apparent reduction in transport of labeled glycoproteins was accounted for by a marked depression in incorporation. The inference from these findings, that monensin acts to block fast transport at the level of the Golgi apparatus, was supported by ultrastructural examination of the drug-treated neurons. An extensive and selective disruption of Golgi saccules was observed, accompanied by an accumulation of clumped smooth membranous cisternae. Quantitative analyses of 48 individual fast-transported protein species, after separation by two-dimensional gel electrophoresis, revealed that monensin depresses all proteins to a similar extent. These results indicate that passage through the Golgi apparatus is an obligatory step in the intracellular routing of materials destined for fast axonal transport.
D000818:Animals; D001370:Axonal Transport / Q000187:drug effects; D005643:Fucose / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D005727:Ganglia, Spinal / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D007930:Leucine / Q000378:metabolism; D008985:Monensin / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D011892:Rana catesbeiana
D006023:Glycoproteins; D009419:Nerve Tissue Proteins; D005643:Fucose; D008985:Monensin; D007930:Leucine; D005990:Glycerol; D005690:Galactose