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The Journal of Cell Biology, Vol 59, 643-660, Copyright © 1973 by Rockefeller University Press

ARTICLE

THE FINE STRUCTURE AND IMMUNOLOGICAL LABELING OF THE ACHROMATIC MITOTIC APPARATUS AFTER DISRUPTION OF CELL MEMBRANES



Samuel Dales 1, Konrad C. Hsu 1, and Ariaki Nagayama 1

1 From the Department of Cytobiology, The Public Health Research Institute of the City of New York, Inc., New York 10016 and the Department of Urology, Columbia University College of Physicians and Surgeons, New York 10032.

Dr. Nagayama's present address is the Department of Microbiology, School of Dentistry, Kyushu University, Fukuoka, Japan.

After treatment of HeLa and L cells with vinblastine sulfate the material of microtubules (tubulin) was reorganized into (a) large paracrystals (PC) of tightly packed tubules; (b) smaller aggregates of tubules with greater diameter whose walls are constituted from well defined, helically arranged morphological subunits; and (c) microtubules associated with helices of polyribosomes of uniform size. All of these structures survived disruption of cellular membranes by means of a nonionic detergent. Following a thorough stripping of membranes there remained a subcellular fraction sedimenting at 1,500 g for 15 min, in which were contained nuclei, centrioles, and the above mentioned microtubular elements, maintained as a complex of organelles by an interconnecting network of 80 Å microfibrils.

As a result of membrane disruption it was possible to localize precisely in the electron microscope the binding of ferritin antibody conjugates. Specific labeling at the surface of PC and microtubule aggregates could be demonstrated. This result was substantiated by means of the immunoperoxidase method of labeling the PC.

A concentrated deposit of ferritin was also found in the vicinity of centrioles and related structures, the annuli of the nuclear pore complex and the annulate lamellae. However, the specificity of the label on these organelles remains questionable because ferritin, albeit in lower concentration, was also present on them in control preparations reacted with preimmune sera.

Submitted on June 22, 1973
Revised on July 23, 1973


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