Fig. 3

Localization of cyclin B1 reporter mRNAs in oocytes of transgenic zebrafish. Whole-mount in situ hybridization of full-grown oocytes probed with cyclin B1 (A) or gfp ((B)?(I)). Shown are oocytes expressing tgo3′ (B), 1?523 (C), 524?1197 (D), 524?736 (E), 737?949 (F), 949?1197 (G), tgo^M3′ (H) and SV40 (I) mRNAs. The tgo32, 524?1197 and 524?736 mRNAs were aggregated in the animal polar cytoplasm ((B), (D), (E)). In contrast, the 1?523, 737?949, 949?1197 and tgo^M3′ mRNAs were dispersed in the animal hemisphere ((C), (F), (G), (H)) and the SV40 mRNAs were dispersed throughout the oocyte (I). Arrows indicate signals of cyclin B1 (A) and reporter mRNAs ((B)?(H)). Bars, 100 μm. (J) Structure of the SV40 reporter gene, which contains cyclin B1 5′ UTR (5′UTR), coding sequences of TC-tag (TC) and EGFP (GFP), a stop codon, cyclin B1 coding region and SV40 3′ UTR (SV40). (K) Quantification of reporter mRNAs (normalized to β-actin mRNA) in full-grown transgenic oocytes by real-time PCR. No statistically significant difference was found in the contents of reporter mRNAs. Error bars indicate mean±s.e.m. (n=3).