Dithiothreitol prevents membrane fusion but not centrosome or microtubule organization during the first cell cycles in sea urchins.
Dithiothreitol (DTT), a disulfide reducing agent, inhibits the fusion of male and female pronuclei within the activated cytoplasm of sea urchin eggs. The migrations of the pronuclei are not affected by DTT, indicating that microtubule function is not impaired. Centrosomal antigens are detected in the sperm aster and in all subsequent microtubule-based configurations. Nuclear membranes never fuse and the chromatin of male and female pronuclei never mix in the DTT-treated cells. During prophase, when nuclear envelopes break down to undergo mitosis, both sets of chromosomes undergo condensation cycles independent from each other. Both pronuclei initially stain for centrosomal material and surrounding microtubules. With time, the female's centrosomal material as well as the microtubules disappear while the male forms a bipolar spindle. Interestingly, one pole of the paternal mitotic apparatus communicates with the separate maternal chromatin, forming a half spindle which moves the egg-derived chromatin towards its pole. At the time for cell division, the individual karyomeres are not able to fuse their nuclear membranes to reconstitute the blastomere nuclei. When DTT is applied at prometaphase of the first cell cycle, the chromosome cycle continues until next metaphase. Centrosomes also continue their cycle and undergo somewhat atypical splitting during the time for second telophase. Division furrows are initiated but aborted. These results support the hypothesis that disulfide groups are required for membrane fusion of the pronuclei, for membrane fusion of the karyomeres, and for the completion of the division furrow to achieve successful cell division.
Schatten
H
H
Department of Zoology, University of Wisconsin, Madison 53706.
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects*; D002455:Cell Division; D003553:Cystine / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology*; D005260:Female; D005306:Fertilization; D008297:Male; D008561:Membrane Fusion / Q000187:drug effects*; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000187:drug effects*; D012617:Sea Urchins / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure; D015053:Zygote / Q000187:drug effects* / Q000378:metabolism
D003553:Cystine; D004229:Dithiothreitol