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PMID: 8320258 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Microtubule nucleation and release from the neuronal centrosome.

The Journal of cell biology ·Vol. 122 ·No. 2 ·1993-07-00 ·Pages 349-59

Yu W, Centonze VE, Ahmad FJ, Baas PW

Abstract

We have proposed that microtubules (MTs) destined for axons and dendrites are nucleated at the centrosome within the cell body of the neuron, and are then released for translocation into these neurites (Baas, P. W., and H. C. Joshi. 1992. J. Cell Biol. 119:171-178). In the present study, we have tested the capacity of the neuronal centrosome to act as a generator of MTs for relocation into other regions of the neuron. In cultured sympathetic neurons undergoing active axonal outgrowth, MTs are present throughout the cell body including the region around the centrosome, but very few (< 10) are directly attached to the centrosome. These results indicate either that the neuronal centrosome is relatively inactive with regard to MT nucleation, or that most of the MTs nucleated at the centrosome are rapidly released. Treatment for 6 h with 10 micrograms/ml nocodazole results in the depolymerization of greater than 97% of the MT polymer in the cell body. Within 5 min after removal of the drug, hundreds of MTs have assembled in the region of the centrosome, and most of these MTs are clearly attached to the centrosome. A portion of the MTs are not attached to the centrosome, but are aligned side-by-side with the attached MTs, suggesting that the unattached MTs were released from the centrosome after nucleation. In addition, unattached MTs are present in the cell body at decreasing levels with increasing distance from the centrosome. By 30 min, the MT array of the cell body is indistinguishable from that of controls. The number of MTs attached to the centrosome is once again diminished to fewer than 10, suggesting that the hundreds of MTs nucleated from the centrosome after 5 min were subsequently released and translocated away from the centrosome. These results indicate that the neuronal centrosome is a highly potent MT-nucleating structure, and provide strong indirect evidence that MTs nucleated from the centrosome are released for translocation into other regions of the neuron.

MeSH Terms
Animals Cells, Cultured Centrioles/metabolism,ultrastructure Ganglia, Sympathetic/cytology Microscopy, Electron Microscopy, Fluorescence Microtubules/chemistry,metabolism,ultrastructure Neurons/chemistry,metabolism,ultrastructure Nocodazole/pharmacology Rats Trimethoprim, Sulfamethoxazole Drug Combination/analysis Tubulin/analysis
Chemicals
Tubulin Trimethoprim, Sulfamethoxazole Drug Combination Nocodazole
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Yu W
Department of Anatomy, University of Wisconsin Medical School, Madison 53706.
Centonze V E
Ahmad F J
Baas P W
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1993-07-00
Pages
349-59
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2119640
Subset
IM
Grants
NINDS NIH HHS · NS 28785 · United States
NCRR NIH HHS · RR00570 · United States
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