ZFIN ID: ZDB-FIG-191230-1484
Duchemin et al., 2019 - Mechanically activated Piezo channels modulate outflow tract valve development through the Yap1 and Klf2-Notch signaling axis. eLIFE   8 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 5
PHENOTYPE:
Fish:
Observed In:
Stage Range: Protruding-mouth to Day 5

Figure 8 Flow and mechanosensitive channels are necessary for proper OFT valve formation. (A) Z-sections and quantifications of the valves phenotypes (normal, thick, delayed) of the Tg(fli:lifeaAct-EGFP; kdrl:nls-mCherry) at 72 hpf (B), 96 hpf (C) and 120 hpf (D) in trpp2+/+(n = 11, n = 10, n = 10), trpp2-/- (n = 8, n = 13, n = 13 from three independent experiments), piezo1+/+ (n = 10, n = 10, n = 9), piezo1-/-(n = 10, n = 10, n = 9 from two independent experiments), piezo1+/+; trpp2+/+ (n = 7), trpp2-morpholino injected embryos (n = 9) and piezo1-/-; trpp2-morpholino injected embryos (n = 7). Scale bar: 10 µm. (D) Z-section of the OFT stained with DAF-FMDA in piezo1+/+ and piezo1-/-. Scale bar: 20 µm. (E) Fibronectin1 (Fn1), elastin (Elnb) and Yap1 staining on Tg(fli:lifeact-eGFP) in piezo1+/+ (n = 12, n = 4 and n = 10 respectively) and piezo1-/- (n = 12, n = 4 and n = 10 respectively from three independent experiments). Scale bar: 20 µm. Z-sections (F) and quantification (F’) of the klf2a reporter (GFP over background) in the anterior and posterior parts of the valves in piezo1+/+ (n = 6) and piezo1-/- (n = 6) obtained from two independent experiments). Scale bar: 20 µm. Z-sections (G) and quantification (G’) of the klf2a reporter (GFP over background) in the anterior and posterior parts of the valves in trpp2+/+ (n = 6) and trpp2-/- (n = 6) (obtained from two independent experiments). Scale bar: 20 µm. Student’s t-test. Boxplot: Center lines show the medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles, outliers are represented by dots.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP ig11Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
ig11Tg ; pkd2tc321/tc321 standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
piezo1sa12608/sa12608; ig11Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
piezo1sa12608/sa12608; zf495Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
y173Tg; zf495Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
Day 4 bulbus arteriosus endocardial cushion IFL
Day 5 ventriculo bulbo valve IFL
zf495Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
mCherry y173Tg; zf495Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion IFL
Day 4 bulbus arteriosus endocardial cushion IFL
Day 5 ventriculo bulbo valve IFL
yap1 piezo1sa12608/sa12608; zf495Tg standard conditions Protruding-mouth bulbus arteriosus vascular smooth muscle IHC
zf495Tg standard conditions Protruding-mouth bulbus arteriosus vascular smooth muscle IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-elnb IHC piezo1sa12608/sa12608; zf495Tg standard conditions Protruding-mouth bulbus arteriosus vascular smooth muscle
IHC zf495Tg standard conditions Protruding-mouth bulbus arteriosus vascular smooth muscle
Ab2-fn IHC piezo1sa12608/sa12608; zf495Tg standard conditions Protruding-mouth bulbus arteriosus vascular smooth muscle
IHC zf495Tg standard conditions Protruding-mouth bulbus arteriosus vascular smooth muscle
Phenotype Details
Fish Conditions Stage Phenotype
ig11Tg ; pkd2tc321/tc321 standard conditions Protruding-mouth bulbus arteriosus endocardial cushion EGFP expression decreased amount, abnormal
Protruding-mouth ventriculo bulbo valve development decreased process quality, abnormal
piezo1sa12608/sa12608; ig11Tg standard conditions Protruding-mouth bulbus arteriosus endocardial cushion EGFP expression increased amount, abnormal
Protruding-mouth ventriculo bulbo valve development decreased process quality, abnormal
piezo1sa12608/sa12608; zf495Tg standard conditions Protruding-mouth bulbus arteriosus smooth muscle tissue development decreased process quality, abnormal
Protruding-mouth bulbus arteriosus vascular smooth muscle ab1-elnb labeling decreased amount, abnormal
Protruding-mouth bulbus arteriosus vascular smooth muscle yap1 expression decreased amount, abnormal
Protruding-mouth bulbus arteriosus vascular smooth muscle ab2-fn labeling decreased amount, abnormal
pkd2tc321/tc321; y173Tg; zf495Tg standard conditions Day 5 ventriculo bulbo valve morphogenesis decreased process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife