ZFIN ID: ZDB-FIG-130423-37
Tay et al., 2013 - Lethal giant larvae 2 regulates development of the ciliated organ Kupffer's vesicle. Development (Cambridge, England)   140(7):1550-1559 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 2-cell to Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 5-9 somites to Long-pec

Fig. 1 Lgl2 controls development of ciliated organs. (A-H) Whole-mount RNA in situ hybridizations. At the 2-cell stage, antisense lgl2 probes show that lgl2 mRNA is maternally supplied (A). Control lgl2 sense probes showed little background staining (B). lgl2 expression localized in the enveloping layer and dorsal forerunner cells (DFCs) at the 80% epiboly stage (C) and was prominently expressed in Kupffer’s vesicle (KV) at the 2-3 somite(s) stages (D,E). At 24 hpf, lgl2 is highly expressed in the ciliated nasal placodes (np), otic vesicles (ov) and pronephric ducts (pnd) (arrows in F). G and H show higher magnification images of lgl2 expression in the nasal placode and otic vesicle (G) and pronephric duct (H). (I-L) Embryo morphology at 2 dpf. Control embryos (I) had a straight body, whereas Lgl2 MO-1 (J) and Lgl2 MO-2 (K) embryos often showed a curved body. Lgl2 MO embryos also developed hydrocephalus (arrows in J-L) and in some cases had additional severe axial defects (L). (M) Percentage of embryos with body axis defects. n, number of embryos analyzed. (N-S) Fluorescence immunostaining using aPKC antibodies (red) to mark apical membranes of epithelial cells and acetylated tubulin antibodies (green) to label cilia revealed defects in the development of ciliated epithelia in KV (N,O), otic vesicles (P,Q) and pronephric ducts (R,S) in Lgl2 MO embryos (O,Q,S) relative to controls (N,P,R). White boxes indicate areas enlarged in ‘Merge & Zoom’ panels. Scale bars: 20 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
llgl2 AB/TU standard conditions 2-cell whole organism ISH
75%-epiboly EVL ISH
75%-epiboly forerunner cell group ISH
1-4 somites Kupffer's vesicle ISH
Prim-5 olfactory placode ISH
Prim-5 otic vesicle ISH
Prim-5 pronephric duct ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-prkca IHC AB/TU control 5-9 somites Kupffer's vesicle apical plasma membrane
IHC Prim-5 otic vesicle apical plasma membrane
IHC Prim-5 pronephric duct apical plasma membrane
IHC AB/TU + MO1-llgl2 standard conditions 5-9 somites Kupffer's vesicle apical plasma membrane
IHC Prim-5 otic vesicle apical plasma membrane
IHC Prim-5 pronephric duct apical plasma membrane
Ab1-tuba IHC AB/TU control 5-9 somites Kupffer's vesicle motile cilium
IHC Prim-5 otic vesicle motile cilium
IHC Prim-5 pronephric duct motile cilium
IHC AB/TU + MO1-llgl2 standard conditions 5-9 somites Kupffer's vesicle motile cilium
IHC Prim-5 otic vesicle motile cilium
IHC Prim-5 pronephric duct motile cilium
Phenotype Details
Fish Conditions Stage Phenotype
AB/TU + MO1-llgl2 standard conditions 5-9 somites Kupffer's vesicle decreased volume, abnormal
5-9 somites Kupffer's vesicle has fewer parts of type Kupffer's vesicle motile cilium, abnormal
5-9 somites Kupffer's vesicle development decreased process quality, abnormal
5-9 somites Kupffer's vesicle motile cilium decreased length, abnormal
5-9 somites Kupffer's vesicle motile cilium assembly decreased process quality, abnormal
Prim-5 otic vesicle decreased volume, abnormal
Prim-5 otic vesicle has fewer parts of type otic vesicle motile cilium, abnormal
Prim-5 otic vesicle motile cilium decreased length, abnormal
Prim-5 otic vesicle motile cilium assembly decreased process quality, abnormal
Prim-5 pronephric duct morphogenesis decreased process quality, abnormal
Prim-5 pronephric duct motile cilium decreased length, abnormal
Prim-5 pronephric duct motile cilium disorganized, abnormal
Long-pec brain hydrocephalic, abnormal
Long-pec post-vent region curved ventral, abnormal
AB/TU + MO3-llgl2 standard conditions Long-pec brain hydrocephalic, abnormal
Long-pec post-vent region curved ventral, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development