Taxol-induced assembly of brain and testis tubulins, and ovarian follicle tubulin dynamics in the frog (genus Rana), in vitro.
1. Tubulins from brain and testes of the frog (genus Rana) were purified by taxol and cyclic assembly-disassembly. 2. Attempts to purify ovarian tubulins by this same method were unsuccessful. 3. Oocyte fractions isolated in microtubule-stabilizing detergent buffer were analyzed by SDS-PAGE and Western blots were probed with monoclonal antibodies to tubulins. 4. In the fully-grown ovarian follicle (diameter approx. 1.7 mm), the majority of tubulin was present in the supernatant fraction. 5. Tubulin was routinely detected in the medium speed (16,000 g) prophase I oocyte pellet. 6. Nocodazole treatment of ovarian follicles in vitro reduced the pelletable tubulin detected, while taxol increased tubulin in the pellet and decreased the supernatant tubulin. 7. These results indicate that taxol is useful in the purification of brain and testes tubulin from Rana tissue; while oocyte tubulin appears to respond to taxol in vivo, another method will be required for in vitro assembly and purification of ovarian tubulin.
Lessman
C A
CA
Department of Biology, Memphis State University, TN 38152.
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D001921:Brain / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008297:Male; D008854:Microscopy, Electron; D008870:Microtubules / Q000378:metabolism; D006080:Ovarian Follicle / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D017239:Paclitaxel / Q000494:pharmacology*; D011898:Ranidae; D013737:Testis / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D014404:Tubulin / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D014404:Tubulin; D017239:Paclitaxel