ZFIN Image: Bontems et al., 2009, Fig. 3

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Figure Caption

Fig. 3 buc mRNA Translation Is Required to Assemble Germ Plasm, and Buc-GFP Localizes to the Balbiani Body

(A–E) Buc is required in the oocyte for germ plasm localization. Wild-type (A and C) and buc^p106re mutant oocytes (B and D) after microinjection of wild-type (A and B) or mutant buc^p106re mRNA (C and D) into stage I zebrafish oocytes. (E) Quantification of dazl mRNA localization from three independent experiments in WT oocytes, uninjected (75% ± 6.0%, n = 63), injected with WT buc (84% ± 14%, n = 67), or mutant buc^p106re mRNA (83% ± 12%, n = 47) compared to buc^p106re mutant oocytes, uninjected (6% ± 3.9%, n = 122), injected with WT buc (26% ± 5.1%, n = 103), or mutant buc^p106re mRNA (7% ± 6.4%, n = 26). Note the rescue of dazl mRNA localization in mutant oocytes after injection of wild-type buc mRNA (B and E). Error bars represent SD. ^***p = 0.0008.

(F–I) Buc needs to be translated to assemble germ plasm. buc^p106re mutant stage I oocytes injected with mutant buc^p106re (F and H) or WT buc mRNA (G and I) with control (+ctrl-MO) (F and G) or translation inhibition morpholino (+buc-MO) (H, I), which inhibited translation of Buc-GFP fusion mRNA and, hence, GFP-fluorescence efficiently in embryos (Figure S5).

(A–D and F–I) Whole-mount in situ hybridization of dazl mRNA (blue). Scale bars: 25 μm.

(J) Quantification of dazl mRNA localization in mutant oocytes injected with mutant buc^p106re mRNA plus ctrl-MO (9%, n = 11) or plus buc-MO (10%, n = 21) and WT mRNA plus ctrl-MO (25%, n = 20) or plus buc-MO (11%, n = 27). Note that the rescue of dazl mRNA localization is blocked by a morpholino-inhibiting buc mRNA translation (I and J).

(K–M) Buc localizes to the Balbiani body. Living WT oocytes injected with mRNA encoding a Buc-GFP fusion (green) at mid (K) and late-stage Ib (L), as well as stage II (M) of oogenesis. Scale bars represent 25 μm.

(N) Model of the role of Buc in early-stage I oocytes. Buc inhibits the premature localization of foxH1, vg1RBP, and buc mRNA at the animal pole (A) and concomitantly recruits the germ plasm RNAs dazl, nanos, and vasa in the Balbiani body at the vegetal pole (V).

Acknowledgments

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