Changes in the organization of tubulin during meiosis in the eggs of the surf clam, Spisula solidissima

Abstract

Polymerized tubulin can be stabilized in Kane's spindle isolation medium (HGL solution), isolated by differential centrifugation and then assayed by colchicine binding activity. In the eggs of the surf clam, Spisula solidissima, the level of particulate tubulin undergoes a series of specific changes during first meiotic division. In either unactivated ("interphase") eggs or metaphase eggs the amount of particulate tubulin was about 13% of the total at 23 degrees C. The amount of particulate tubulin decreased shortly after activation, reaching a minimum value at about 5 min, the time of nuclear membrane breakdown. The particulate tubulin concentration then rose, reaching a maximum at metaphase, and then decreased again during anaphase, reaching a minimum at first polar body formation. In HGL homogenates of unactivated eggs a structure is present which has been shown to contain the interphase particulate tubulin (IPT). This structure consists essentially of a 10-20 micro granular sphere attached to a membranous material which is probably part of the egg cortex. These particles are absent at the time of nuclear membrane breakdown, when the level of particulate tubulin is minimal and when the first signs of spindle formation are visible. Electron microscopy of these particles by negative staining indicates that they are composed of microtubules associated with a granular matrix which may be a polymorphic aggregate of tubulin.