FIGURE SUMMARY
Title

Zebrafish Numb homologue: Phylogenetic evolution and involvement in regulation of left-right asymmetry

Authors
Niikura, Y., Tabata, Y., Tajima, A., Inoue, I., Arai, K.I., and Watanabe, S.
Source
Full text @ Mech. Dev.

Expression of znumb, notch1a, deltaD in zebrafish development. (A) Semi-quantitative RT-PCR analysis of znumb mRNA expression. Total RNA was isolated from various tissues of adult zebrafish, and znumb transcripts were amplified by using specific primers. μ-actin was used as a control. (B) Whole-mount in situ hybridization patterns of znumb expression during zebrafish embryonic development. Zebrafish embryos at various stages as indicated were hybridized with the antisense probe. Upper (3 hpf) panels are lateral views with the animal pole at the top. For the second (12 hpf) and third (15 hpf) rows, left panels are lateral view of embryos with anterior to the left, middle panels are frontal view with dorsal at the top, and right panels are a view of embryos from the posterior with dorsal at the top. A panel of 20 hpf in the fourth row is a lateral view with anterior to the left. Enlarged image is shown to the right side. (C,D) Whole-mount in situ hybridization patterns of notch1a (C) and deltaD (D). Left panels are lateral view of embryos with anterior to the left, middle panels are frontal view with dorsal at the top, and right panels are views from caudal to anterior with dorsal at the top. nk, neural keel; t, telencephalon; d, diencephalon; m, midbrain; h, hindbrain; kv, Kupffer′s vesicle; s, somites; tb, tail bud; nr, neural retina; sc, spinal cord. Sense probe hybridization was done for all samples, and no significant signals were observed (data not shown).

Whole-mount in situ analysis of left–right asymmetry markers. Expression of bmp4 (A), nkx2.5 (B) lefty2 (C), no tail (D) and shh (E) in control (gal4) and znumb mRNA-injected embryos. In C, deltaD and NICD injected embryos were also examined. Zebrafish fertilized eggs were injected with the indicated mRNA, and embryos were hybridized with anti-sense probes for bmp4 (A), nkx2.5 (B) and lefty2 (C) at around 32 hpf and for no tail and shh at around 9–10 hpf. Injected mRNAs are indicated in each panel, and 2 representative samples of znumb-mRNA-injected embryos are shown in A, B and C. Panels of A, B and C are dorsal views of head portion, and arrowheads indicate signals. In D and E, dorsal views with animal pole to top are shown. Sense probe hybridization was done for all samples, and no significant signal was observed (data not shown).

Acknowledgments
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Reprinted from Mechanisms of Development, 123(5), Niikura, Y., Tabata, Y., Tajima, A., Inoue, I., Arai, K.I., and Watanabe, S., Zebrafish Numb homologue: Phylogenetic evolution and involvement in regulation of left-right asymmetry, 407-414, Copyright (2006) with permission from Elsevier. Full text @ Mech. Dev.