ZFIN ID: ZDB-FIG-191230-769
Mano et al., 2019 - Brain-specific homeobox Bsx specifies identity of pineal gland between serially homologous photoreceptive organs in zebrafish. Communications biology   2:364 Full text @ Commun Biol
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage Range: Protruding-mouth to Days 7-13
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Fig. 2

MO-mediated knockdown of bsx. a Schematic illustration of the target site of bsx MO. b RT-PCR of bsx mRNA in 48-hpf embryos injected with control or bsx MO. Full images of the electrophoreses are shown in Supplementary Fig. 8. c Apparent views of 3.5 dpf larvae injected with control or bsx MO. No gross morphological difference was detected. dk Effects of bsx MO on the pineal gland. All the larvae are viewed dorsally with anterior to the top. Three images in each row (d, f, h) were taken from the same larva at different stages. d Size reduction of the pineal gland in bsx morphants. White dashed lines indicate the position of the pineal gland. f Reduction of pineal EGFP fluorescence in Tg(exorh:egfp) by Bsx depletion. h Induction of EGFP expression in Tg(rho:egfp) by Bsx depletion (indicated by white arrowheads). e, g, i Quantification of pineal size and fluorescence signals in bsx MO- or control MO-injected larvae. Horizontal bars indicate mean values. The difference between bsx MO-injected and control groups was statistically significant for each case [P < 2.2 × 10−16 (3.5, 6.5, and 9.5 dpf, e), P = 2.8 × 10−12 (3.5 dpf, g), P < 2.2 × 10−16 (6.5 and 9.5 dpf, g), P = 2.0 × 10−8 (3.5 dpf, i), P = 1.3 × 10−7 (6.5 dpf, i), P = 0.011 (9.5 dpf, i) by Welch’s two-sided t-test]. j Effects of bsx MO on Tg(exorh:egfp);Tg(rho:ntr-mCherry) double reporter line at 5.5 dpf. In control larva, EGFP (green) and NTR-mCherry (magenta) were localized in the pineal gland and the retina, respectively. In bsx morphant, both EGFP (green) and NTR-mCherry (magenta) signals were detected in the pineal gland. k A confocal image of pineal cells in Bsx-depleted Tg(exorh:egfp);Tg(rho:ntr-mCherry), showing mutually exclusive expression of EGFP (green) and NTR-mCherry (magenta). Scale bars, 1 mm (c), 30 μm (d, f), 100 μm (h,j), 10 μm (k)

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP ja1Tg standard conditions Protruding-mouth epiphysis IFL
Day 6 epiphysis IFL
Days 7-13 epiphysis IFL
ja1Tg + MO1-bsx standard conditions Protruding-mouth epiphysis IFL
Day 6 epiphysis IFL
Days 7-13 epiphysis IFL
ja1Tg; ja83Tg control Day 5 epiphysis IFL
ja1Tg; ja83Tg + MO2-bsx standard conditions Day 5 epiphysis IFL
ja2Tg control Protruding-mouth Not Detected epiphysis IFL
Day 6 Not Detected epiphysis IFL
ja2Tg + MO1-bsx standard conditions Protruding-mouth epiphysis IFL
Day 6 epiphysis IFL
mCherry ja1Tg; ja83Tg control Day 5 Not Detected epiphysis IFL
ja1Tg; ja83Tg + MO2-bsx standard conditions Day 5 epiphysis IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-bsx standard conditions Protruding-mouth epiphysis decreased area, abnormal
Protruding-mouth epiphysis decreased size, abnormal
Day 6 epiphysis decreased area, abnormal
Day 6 epiphysis decreased size, abnormal
Days 7-13 epiphysis decreased area, abnormal
Days 7-13 epiphysis decreased size, abnormal
ja1Tg; ja83Tg + MO2-bsx standard conditions Day 5 epiphysis EGFP expression increased amount, abnormal
Day 5 epiphysis mCherry expression increased amount, abnormal
ja2Tg + MO1-bsx standard conditions Protruding-mouth epiphysis EGFP expression increased amount, abnormal
Day 6 epiphysis EGFP expression increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Communications biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Commun Biol