Alkaline pH favors microtubule self-assembly in surf clam, Spisula solidissima, oocyte extracts.
Microtubule assembly in surf clam oocytes is dependent upon events that occur during fertilization. Prior to fertilization there are few, if any microtubules, but within minutes after fertilization microtubules assemble to form the meiotic apparatus. This study demonstrates that the assembly of microtubules after fertilization may be dependent on the fertilization-induced pH change of the cytoplasm. Since the magnitude of the intracellular pH (pHi) change in Spisula oocytes has not been determined, surf clam microtubule assembly was examined at pH values that reflect the pHi change that occurs during sea urchin fertilization. The results indicate that microtubule assembly in crude oocyte extracts is favored at alkaline pH. In contrast, purified surf clam tubulin assembles to a greater extent at pH 6.6 than at pH 7.2. These results reveal that the tubulin in unfertilized oocytes can assemble into microtubules at pH 6.6 but that they are prevented from doing so by pH-dependent cytoplasmic regulatory factors in the oocyte.
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Marine Biological Laboratory, Woods Hole, Massachusetts 02543.
D000818:Animals; D002474:Cell-Free System; D003593:Cytoplasm / Q000502:physiology / Q000648:ultrastructure; D005306:Fertilization; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D008970:Molecular Weight; D008974:Mollusca; D009865:Oocytes; D011485:Protein Binding; D014404:Tubulin / Q000378:metabolism*
D008869:Microtubule-Associated Proteins; D014404:Tubulin