Microtubule tracks can be detected in mouse oocytes with an antibody directed against a calcium transporter.
In metaphase II-arrested mouse oocytes, most microtubules are found in the meiotic spindle, a structure that remains stable for hours despite microtubule instability. Microtubule organizing centres (MTOCs) are present at the poles of the spindle and in the cytoplasm, but the latter nucleate very few microtubules. This particular organization of the microtubule network enabled us to observe the unexpected behaviour of a protein that can associate with microtubules. We compared the distribution of a mitosis-activated calcium transport system with that of the microtubule network, by immunofluorescence, using two monoclonal antibodies, one directed against a component of the calcium transport system (7/13), and the other against the common tyrosinated form of alpha-tubulin (YL1/2). The 7/13 staining was associated with the spindle microtubules and with the kinetochore area. In addition, we observed many asters in the cytoplasm, around the cytoplasmic MTOCs. The majority of these asters were not stained with the antitubulin antibody. Moreover, these 7/13 asters either disappeared after nocodazole treatment or were enlarged after taxol treatment. Using a confocal microscope, we observed single fibres that were stained with both antibodies: the extremity furthest from the MTOC (corresponding to the + end of the microtubule) being detected by the 7/13 antibody only. All these observations suggest that the 7/13 antigen is associated with microtubule tracks that persist a few minutes after microtubule depolymerization. The possible role of these tracks in microtubule regrowth is discussed.
de Pennart
H
H
Département de Biologie du Développement, Institut Jacques Monod, CNRS-Université Paris 7, France.
Cibert
C
C
Petzelt
C
C
Maro
B
B
D000818:Animals; D000906:Antibodies; D015415:Biomarkers / Q000032:analysis; D002135:Calcium-Binding Proteins / Q000032:analysis*; D003599:Cytoskeleton / Q000648:ultrastructure; D005260:Female; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D008540:Meiosis; D008677:Metaphase; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D017239:Paclitaxel / Q000494:pharmacology
D000906:Antibodies; D015415:Biomarkers; D002135:Calcium-Binding Proteins; D017239:Paclitaxel