FIGURE SUMMARY
Title

Brorin is required for neurogenesis, gliogenesis, and commissural axon guidance in the zebrafish forebrain

Authors
Miyake, A., Mekata, Y., Fujibayashi, H., Nakanishi, K., Konishi, M., Itoh, N.
Source
Full text @ PLoS One
Fig. 2

Pattern of brorin expression in zebrafish embryos.

(A) Amplification of brorin by RT-PCR at the indicated stages (the lower panel shows ef1α as a control). (B-K) Expression of brorin in zebrafish embryos at the indicated stages as detected by whole-mount in situ hybridization. B-D, F, H, and J are lateral views, anterior to the left; E is the ventral view; G and I are dorsal views; K is the frontal view. Arrows in panels D and H indicate the pituitary gland. Arrows in panels E and I indicate the olfactory placode. ac, anterior commissure; di, diencephalon primordium; hb, hindbrain; hy, hypothalamus; mb, midbrain; p, prethalamus; pt, posterior tuberculum; te, telencephalon.
Fig. 3

Inhibition of brorin function in zebrafish embryos.

(A) The coding region of brorin is divided into two introns, with open boxes and black lines indicating exons and introns, respectively. MO indicates the target position of brorin MO. (B) brorin cDNA was amplified from the cDNA of wild-type embryos or brorin MO-injected embryos by RT-PCR using the P1 and P2 primers, the positions of which are indicated by arrows (A). (C) The nucleotide sequences of brorin cDNAs were elucidated. Numbers show the nucleotide sequence of the coding region and amino acid sequence, and arrowheads indicate splice sites between exons one and two. (D-G) Lateral views of control MO-injected (D), brorin MO1-injected (E), brorin MO2-injected (F), and brorin MO1- and brorin RNA-injected (G) embryos at 24 hpf. Arrows and arrowheads indicate the tectal ventricle and telencephalic ventricle, respectively. di, diencephalon; mb, midbrain; te, telencephalon.
Fig. 4

pSmad distribution and expression of the Wnt target gene in brorin morphants.

(A, B) Pattern of pSmad expression in wild-type embryos (A) and brorin morphants (B) at 24 hpf. (C, D) Expression of axin2 in wild-type embryos (C) and brorin morphants (D) at 24 hpf. Lateral view with the anterior to the left.

EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Term:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Prim-5
Fig. 5

pSmad distribution in brorin RNA-injected embryos.

(A-D) Lateral views of wild-type (A, C) and brorin RNA-injected (B, D) embryos at 24 hpf. (E, F) Pattern of pSmad expression in wild-type (E) and brorin RNA-injected (F) embryos at 8 hpf. (G-J) Pattern of pSmad expression in wild-type (G, I) and brorin RNA-injected (H, J) embryos at 24 hpf. G and H are optical cross-sections; I and J are lateral views anterior to the left. Scale bar: 50 μm.
Fig. 6

Telencephalic and diencephalic gene expression in brorin morphants.

(A-D) The expression of emx1 (A, B) and tbr1 (C, D) in wild-type embryos (A, C) and brorin morphants (B, D) at 24 hpf. Arrows in panels B and D indicate the ectopic expression of emx1 or tbr1 in the subpallial domain of the telencephalon. (E-H) The expression of dlx2a in wild-type embryos (E, G) and brorin morphants (F, H) at 8 (G, H) and 24 (E, F) hpf. (I, J) The expression of shh in wild-type embryos (I) and brorin morphants (J) at 24 hpf. hy, hypothalamus; p, prethalamus; pa, pallial telencephalon; sp, subpallial telencephalon.
Fig. 7

Specification of GABAergic interneurons and oligodendrocytes, and astroglial development in brorin morphants.

The expression of ascl1a (A, B), gad1 (C, D), ngn1 (E, F), plp (G, H), and glula (I, J) in wild-type embryos (A, C, E, G, I) and brorin morphants (B, D, F, H, J) is displayed at the indicated stages. di, diencephalon; pa, pallial telencephalon; poc, postoptic commissure; sp, subpallial telencephalon.

EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Day 4
Fig. 8

Defects in axon guidance in brorin morphants.

Fluorescent immunolabeling of axons (αAT) in wild-type embryos (A, C) and brorin morphants (B, D) at 28 hpf. The arrow and arrowhead in panel A indicate the AC and POC, respectively. The arrow in panel C indicates the SOT. A and B are frontal views; C and D are lateral views, with the anterior to the left.
Fig. 9

Expression of axon growth substrates in brorin morphants.

Expression of netrin1a (A, B) and sema3d (C, D) in wild-type embryos (A, C) and brorin morphants (B, D) at 24 hpf.

EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Prim-5
Fig. S1

Effects of brorin knockdown on the specification of GABAergic interneurons, oligodendrocytes, and astrocytes, and axon guidance.

The expression of gad1 (A, B), plp (C, D), glula (E, F), and netrin1a (G, H) in brorin MO2-injected (A, C, E, G) and brorin MO1- and brorin RNA-injected (B, D, F, H) embryos is displayed at the indicated stages.
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. Full text @ PLoS One
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