FIGURE

Fig. 9

ID

ZDB-FIG-210414-78

Publication

Takahashi et al., 2020 - A globin-family protein, cytoglobin 1, is involved in the development of neural crest-derived tissues and organs in zebrafish

Other Figures

All Figure Page

Back to All Figure Page

Fig. 9

Knockdown of cygb1 phenocopied the kzt mutation. (A) Morpholino oligos were designed against the translational start codon (cygb1-MO1) and the donor site at the exon 1-intron 1 junction (cygb1-MO2). As a negative control, cygb1-MO1 with five mismatches was used in parallel (5mis-MO). (B) Embryos injected with 5mis-MO at 1 pmol/embryo (a, b) and cygb1-MO1 at 0.04 pmol/embryo (c, d), 0.2 pmol/embryo (e), and 1 pmol/embryo (f) were observed at 28 hpf. cygb1-MO1 at 0.04 pmol/embryo caused head flattening (two-headed arrows), small eyes (two-headed broken arrows), trunk-tail bending (arrowhead), and heart edema (arrow), thus phenocopying the kzt mutation. A higher concentration (1 pmol/embryo) resulted in phenotypes much more severe than those of kzt embryos, whereas 5mis-MO1 resulted in few anomalies. (C) Dose-dependent effects of cygb-MO1 on development. Embryos with normal morphology, kzt-phenotypes, amorphous embryos, and death were scored at different doses of cygb1-MO1. (D) Development of the cranial cartilage in embryos injected with morpholino oligos. Cranial cartilage was stained with Alcian blue in embryos injected with 5mis-MO (a, a’) or cygb1-MO1 (b, b’). (E) Knockdown of cygb1 resulted in molecular effects in embryos similar to those of the kzt mutation. The expression of myod in adaxial cells and crestin in NCCs was examined in embryos injected with 5mis-MO (a–c), cygb1-MO1 (a’–c’), or cygb1-MO2 (a’’, b’’). The pattern of myod expression was particularly affected in the anterior adaxial cells (arrows), and crestin expression was particularly decreased in pharyngeal arch-derived cartilage (brackets). (B, Da, Db) Lateral views with anterior to the left and dorsal to the top. (Da’, Db’, E) Dorsal views with anterior to the left. Percentages of abnormal embryos and the numbers of embryos examined are shown at the bottom right or at the top right. ac, adaxial cell; ch, ceratohyal; cb, ceratobranchial; cnc, cranial neural crest cell; h, heart; hb, hindbrain; hm, hyomandibular; mb, midbrain; mc, Meckel’s cartilage; pa, pharyngeal arch; t, trabecula, te, telencephalon. Scale bars, 200 μm (B, E) or 100 μm (D).

Expression Data

Genes:	crestin myod1 nkx2.5
Fish:	RW RW + MO1-cygb1 RW + MO2-cygb1
Knockdown Reagents:	MO1-cygb1 MO2-cygb1
Anatomical Terms:	head heart tube paraxial mesoderm paraxial mesoderm anterior region pharyngeal arch
Stage Range:	1-4 somites to Prim-5

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	RW + MO1-cygb1 RW + MO2-cygb1
Knockdown Reagents:	MO1-cygb1 MO2-cygb1
Observed In:	axis ceratobranchial cartilage eye head Meckel's cartilage paraxial mesoderm anterior region pericardium trabecula cranii
Stage Range:	1-4 somites to Day 4

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image.

Reprinted from Developmental Biology, 472, Takahashi, K., Ito, Y., Yoshimura, M., Nikaido, M., Yuikawa, T., Kawamura, A., Tsuda, S., Kage, D., Yamasu, K., A globin-family protein, cytoglobin 1, is involved in the development of neural crest-derived tissues and organs in zebrafish, 1-17, Copyright (2020) with permission from Elsevier. Full text @ Dev. Biol.