This proposal deals with the study of the mitotic spindles of invertebrates zygotes. The major goal is to purify tektin-like proteins from mitotic spindles and to determine how these proteins interact with the microtubules of the spindle and the spindle matrix. Tektin antibodies will be used to identify and purify similar proteins from the spindles and these proteins will be used to generate antibodies to be used as specific probes. Laser confocal light microscopy and transmission and scanning electron microscopy will be used to determine the 3-dimensional organization of these matrix polypeptides. Antibodies will be microinjected into living cells to determine effects on spindle structure and function. These studies will advance our understanding of microtubule assembly, polarity and stability in the mitotic spindle and how the matrix provides a framework for the operation of the cell division process.
