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motley/birc5b mutants fail to effectively multimerize GP RNPs. Animal views of blastodisc cortex. In wild-type embryos (A–D) the GP RNP aggregation wave (bracket, A) coincides with cortical astral microtubule ends (B, bracket indicates the wavefront within the wave). GP RNP multimerization in wild-type embryos result in large GP RNP aggregates (C, D). In motley/birc5b mutants (E–H), cortical microtubules are disorganized (F), and GP RNPs fail to multimerize effectively (G, H). In nocodazole-treated embryos (I–L), cortical microtubules are absent (J) and GP RNP multimerization also fails (K, L). Semi-quantitative analysis of GP RNP aggregation showing similar multimerization failures in motley/birc5b and nocodazole-treated embryos compared to wild-type (M). Inset labels in panels A, E, I and C, G, and K indicate ROIs shown at higher magnifications next to the respective panels. Arrows in A, E and I indicate the radial direction, from the center of the blastodisc, along which microtubules in B tend to be oriented in wild-type embryos. (N–R) consecutive confocal sections acquired with a z-step of 0.5 microns: z5 (N, most cortical), z6 (O) and z7 (P, Q, most internal). Yellow arrow shows a single microtubule observed in Z6 (O), Z7 (P) and more internal (not shown) planes but not in z5 (N), which appears associated with an RNP in a multimerized aggregate (Q, R). White arrow indicates a pair of microtubules that appear to converge on a different set of RNPs in the same aggregate (Q,R). The merge panel in R corresponds to microtubules and RNPs for z7 to show the association of microtubule tips with RNP aggregates.
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