Identification of an axolemma-enriched fraction from peripheral nerve.
A method has been devised for the fractionation of whole peripheral nerve. The procedure utilizes differential centrifugation and separation on a linear sucrose gradient (10-40%, wt/wt). A membrane fraction localized between 26% and 29% sucrose was not only enriched for the plasma membrane markers, 5'-nucleotidase and acetylcholinesterase (AChE), but also possessed the highest binding of [3H]saxitoxin, a specific marker for sodium channels. Neurons in the lumbar dorsal roots and ventral horns of rats were injected with [3H]fucose to label glycoproteins associated with the axolemma from sciatic nerve. Fractionation of the labeled nerves demonstrated a coincidence in the distribution of [3H]fucose-labeled material and AChE activity in the sucrose density gradient. The increase in the specific activity of marker enzymes for plasma membrane, sodium channels, and labeled membrane, previously demonstrated to be of axolemmal origin, identified the 26-29% region of the sucrose gradient as enriched for axolemma derived from peripheral nerve.
D015720:5'-Nucleotidase; D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D001369:Axons / Q000378:metabolism / Q000648:ultrastructure*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D005643:Fucose / Q000378:metabolism; D009708:Nucleotidases / Q000378:metabolism; D051381:Rats; D012530:Saxitoxin / Q000378:metabolism; D012584:Sciatic Nerve / Q000648:ultrastructure*
D005643:Fucose; D012530:Saxitoxin; D000110:Acetylcholinesterase; D009708:Nucleotidases; D015720:5'-Nucleotidase