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Volume 16, Number 19, Issue of October 1, 1996 pp. 5967-5978
Copyright ©1996 Society for NeuroscienceProtein Synthesis within Dendrites: Glycosylation of Newly Synthesized Proteins in Dendrites of Hippocampal Neurons in Culture
Received June 5, 1996; revised July 9, 1996; accepted July 11, 1996.
Enrique R. Torre and Oswald Steward Department of Neuroscience, University of Virginia School of Medicine, Charlottesville, Virginia 22908
There is increasing evidence that certain mRNAs are present in dendrites and can be translated there. The present study uses two strategies to evaluate whether dendrites also possess the machinery for protein glycosylation. First, precursor labeling techniques were used in conjunction with autoradiography to visualize glycosyltransferase activities that are characteristic of the rough endoplasmic reticulum (RER) (mannose) or the Golgi apparatus (GA) (galactose and fucose) in dendrites that had been separated from their cell bodies and in intact neurons treated with brefeldin A or low temperature. Second, immunocytochemical techniques were used to define the subcellular distribution of proteins that are considered markers of the RER (ribophorin I) and GA (p58,
-mannosidase II, galactosyltransferase, and TGN38/41). Autoradiographic analysis revealed that isolated dendrites incorporated sugar precursors in a tunicamycin-sensitive and protein synthesis-dependent manner. Moreover, when intact neurons were pulse-labeled with 3H-labeled sugars at low temperature or after treatment with brefeldin A, labeling was distributed over proximal and sometimes distal dendrites. Immunolabeling for RER markers was predominantly localized in cell bodies but extended for a considerable distance into dendrites of all neurons. Immunolabeling for GA markers was confined to the cell body in ~70% of the neurons, but in 30% of the neurons, the staining extended into proximal and middle dendrites. These results indicate that the machinery for glycosylation extends well into dendrites in many neurons.
Key words: rough endoplasmic reticulum; Golgi apparatus; TGN; dendrites; glycosylation; hippocampal neurons; dendritic RNA
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