Acute inactivation of MAP1b in growing sympathetic neurons destabilizes axonal microtubules.
Microtubule-associated-protein 1b (MAP1b) is abundant in neurons actively extending axons. MAP1b is present on microtubules throughout growing axons, but is preferentially concentrated on microtubule polymer in the distal axon and growth cone. Although MAP1b has been implicated in axon growth and pathfinding, its specific functions are not well understood. Biochemical and transfection studies suggest that MAP1b has microtubule-stabilizing activity, but recent studies with neurons genetically deficient in MAP1b have not confirmed this. We have explored MAP1b functions in growing sympathetic neurons using an acute inactivation approach. Neurons without axons were injected with polyclonal MAP1b antibodies and then stimulated to extend axons. Injected cells were compared to controls in terms of axon growth behavior and several properties of axonal microtubules. The injected antibodies rapidly and quantitatively sequestered MAP1b in the cell body, making it unavailable to perform its normal functions. This immunodepletion of MAP1b had no statistically significant effect on axon growth, the amount of microtubule polymer in the axon, and the relative tyrosinated tubulin content of this polymer, and this was true in sympathetic neurons from rat, wild type mice, and tau knockout mice. Thus, robust axon growth can occur in the absence of MAP1b alone or both MAP1b and tau. However, immunodepletion of MAP1b significantly increased the sensitivity of microtubules in the distal axon and growth cone to nocodazole-induced depolymerization. These results indicate that MAP1b has microtubule-stabilizing activity in growing axons. This stabilizing activity may be required for some axonal functions, but it is not necessary for axon growth.
Tint
Irina
I
Department of Anatomy and Cell Biology, Temple University School of Medicine, Philadelphia, PA 19140, USA. mark.black@temple.edu
Fischer
Itzhak
I
Black
Mark
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001369:Axons / Q000502:physiology*; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D005728:Ganglia, Sympathetic / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008845:Microinjections; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism; D009474:Neurons / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D014404:Tubulin / Q000032:analysis
D000911:Antibodies, Monoclonal; D008869:Microtubule-Associated Proteins; D014404:Tubulin; C085284:microtubule-associated protein 1B