Rapid turnover of microtubule-associated protein MAP2 in the axon revealed by microinjection of biotinylated MAP2 into cultured neurons

Abstract

We studied the mechanism of compartmentation of microtubule-associated protein 2 (MAP2) in the dendrites and cell bodies by using microinjection of biotin-labeled MAP2 into mature spinal cord neurons in culture. MAP2 molecules microinjected into the nerve cell body were distributed not only throughout the cytoplasm of the cell body and dendrites, but also in the axon as far as a few millimeters from the cell body within 24 hr after injection. However, when injected cells were incubated for more than 3 days, the amount of biotin-labeled MAP2 in the axon decreased remarkably compared with that in the dendrites. This indicates that there is no sorting mechanism in the cell body for the transport of MAP2 selectively into the dendrites but that the turnover rate of MAP2 in the axons differs from that in the dendrites. To further characterize the mechanism of MAP2 compartmentation, we performed immunoelectron microscopy of injected cells and detegent extraction of microinjected cells prior to immunocytochemistry with anti-biotin. The results strongly suggest that a large part of axonal MAP2 is not associated with cytoskeleton and that this weak association of MAP2 favors selective loss of MAP2 from the axon.