Expression pattern of lhx9 in zebrafish embryogenesis and retinogenesis. (A–G′) lhx9 expressed in the zebrafish embryo at 24 hpf, 48 hpf, 60 hpf, 72 hpf, 96 hpf, and 5 dpf. t, telencephalon; E, eye; e, epiphysis; d, diencephalon; m, mesencephalon; mhb, mid-hindbrain boundary; hb, hindbrain; ht, heart; pf, pectoral fin; asterisk, hypothalamus; Scale bar, 500 μm. (H1–H8) The expression of lhx9 in the zebrafish retina at 24 hpf, 36 hpf, 48 hpf, 60 hpf, 72 hpf, 96 hpf, 5 dpf, and 6 dpf. Figures are horizontal sections along the temporal-nasal axis (T–N). ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; OPL, outer plexiform layer; IPL, inner plexiform layer. Scale bar=20 μm.

Identification of the efficiency of lhx9 MO in the zebrafish embryo. (A) Schematic diagram of lhx9 MO blocking Lhx9 mRNA splicing and protein translation. Exon 2 was deleted in the lhx9 transcript by MO, the LIM1 domain of Lhx9 protein was removed, and the LIM2 and HD domain can't be assembled because of the wrong amino acid sequences and the early stop codon. (B) RT-PCR confirming the effect of splice-blocking MO on lhx9 transcript at 24 hpf, 48 hpf, 60 hpf, 72 hpf, and 96 hpf. MO lanes (MO cDNA as the template) show two PCR bands, band 1 and 2 (736 bp). (C) Sequence alignment of band 1 (939 bp) and band 2 (736 bp).

Immunofluorescence staining analysis of the retinal neuronal differentiation in the INL. All figures are horizontal sections along the temporal- nasal axis (T-N). (A–D″) Immunofluorescence staining with different ACs markers, GABA, Parvalbumin, Calretinin, and TH, in WT, CT, and MO retinas at 96 hpf. (a-d) statistical analysis of GABA⁺, Parvalbumin⁺, Calretinin⁺ and TH⁺ cells in WT, CT, and MO retinas at 96 hpf. Blue, DAPI staining of the nuclei. Scale bar=20 μm. (a–d) Statistical analysis of GABA⁺, Parvalbumin⁺, Calretinin⁺, and TH⁺ cells in WT (n=16), CT (n=15), and MO (n=16) retinas at 96 hpf. Results are represented as the mean±s.e.m. ns, P>0.05; assessed by one-way ANOVA followed by Tukey's multiple comparisons.

Response of the zebrafish larvae to the light stimulus at 5 dpf. (A) The average swimming speed of the larvae during the last 2 min dark and 2 min light period. (B) The swimming speed during the last 2 min dark and the first 10 s of the 2 min light period. Results are represented as the mean±s.e.m. (WT, n=35; CT, n=34; MO, n=32). **P<0.01; assessed by one-way ANOVA followed by Tukey's multiple comparisons.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.