Limits...
Yes-associated protein (Yap) is necessary for ciliogenesis and morphogenesis during pronephros development in zebrafish (Danio Rerio).

He L, Xu W, Jing Y, Wu M, Song S, Cao Y, Mei C - Int. J. Biol. Sci. (2015)

Bottom Line: We examined expression of Yap during zebrafish embryogenesis, and its transcripts were detected in pronephric duct, while Yap protein was found to be localized in the cytoplasm and apical membrane in kidney epithelium cells.With transgenic Tg(Na(+)/K(+) ATPase:EGFP), we found that lacking Yap led to expansion and discontinuities of pronephric duct, as well as disorganization of cloaca during pronephros morphogenesis.Taken together, our data reveals that Yap is required for pronephric duct integrity, maintenance of baso-lateral cell polarity, and ciliogenesis during zebrafish kidney development.

View Article: PubMed Central - PubMed

Affiliation: 1. Kidney Institute of CPLA, Division of Nephrology, Changzheng Hospital, Second Military Medical University, NO.415 Fengyang Road, Shanghai 200003, China.

ABSTRACT
The Hippo signaling pathway and its transcriptional co-activator Yap are known as essential regulators for cell proliferation and organ size. However, little is known about their roles in kidney development and ciliogenesis. We examined expression of Yap during zebrafish embryogenesis, and its transcripts were detected in pronephric duct, while Yap protein was found to be localized in the cytoplasm and apical membrane in kidney epithelium cells. By morpholino (MO) knockdown of yap expression in zebrafish, the injected larve exhibits pronephic cysts and many aspects of ciliopathy, which can be rescued by full-length yap mRNA, but not yap (S127A) mRNA. With transgenic Tg(Na(+)/K(+) ATPase:EGFP), we found that lacking Yap led to expansion and discontinuities of pronephric duct, as well as disorganization of cloaca during pronephros morphogenesis. Mis-located Na(+)/K(+) ATPase and ciliary abnormalities are also detected in pronephric duct of yap morphants. In addition, genetic analysis suggests that yap interacts with ift20, ift88 and arl13b in pronephric cyst formation. Taken together, our data reveals that Yap is required for pronephric duct integrity, maintenance of baso-lateral cell polarity, and ciliogenesis during zebrafish kidney development.

No MeSH data available.


Related in: MedlinePlus

yap morphants have ciliary defects. (A) In the eye, the outer segment of photoreceptor cell, which is a modified cilium, is hard to detect in the morphants at 3 d.p.f (the left arrow indicates the control, the right arrow shows defect in yap morphant). Bar: 25 μm (B) Compared with the injection control, cilia defects are observed in medial and posterior segment of the pronephric duct in the morphant. Bar: 10 μm. (C) Statistical analysis of the single cilia number per high power field and single cilia length in control and morphant embryos. The cilia are shown by staining with antibody against acetylated α-tublin. α-tub, anti-acetylated α-tublin; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; MD, medial segment of pronephric duct; PD, posterior segment f pronephric duct; MC, multicilia; SC, single cilia; HPF, high power field.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4495411&req=5

Figure 5: yap morphants have ciliary defects. (A) In the eye, the outer segment of photoreceptor cell, which is a modified cilium, is hard to detect in the morphants at 3 d.p.f (the left arrow indicates the control, the right arrow shows defect in yap morphant). Bar: 25 μm (B) Compared with the injection control, cilia defects are observed in medial and posterior segment of the pronephric duct in the morphant. Bar: 10 μm. (C) Statistical analysis of the single cilia number per high power field and single cilia length in control and morphant embryos. The cilia are shown by staining with antibody against acetylated α-tublin. α-tub, anti-acetylated α-tublin; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; MD, medial segment of pronephric duct; PD, posterior segment f pronephric duct; MC, multicilia; SC, single cilia; HPF, high power field.

Mentions: Defects in ciliary genes often lead to pronephric cyst, curved tails, heart edema, smaller eyes in zebrafish as yap morphants exhibit. To examine the effect of Yap in ciliogenesis, we stained the eyes and pronephric ducts of yap morphants and control embryos with acetylated α-tublin. In zebrafish, the outer segment of the photoreceptor is a highly modified cilium giving access to visual transduction, it begins to form by 54 h.p.f and persists thereafter. We found that the outer segments were almost discernable in smaller eyes of 3 d.p.f yap morphants (Fig.5A; arrows). The pronephric duct features multiciliated cells (MCCs) and single ciliated cells (SCCs), which form clustered cilia bundles and single cilia respectively. The single cilia are observed as early as 20 h.p.f while the bundling cilia can only be detected from about 36 h.p.f. We examined cilia formation at 3 d.p.f in at least 7 embryos each group, and found that the MCCs in the mid region of the duct did not form normal bundles but only disordered cilia, which had also been seen in other zebrafish cilia mutants 9, 39, while in the posterior segments, the number (20.3 ± 1.5 in morphant versus 30.3 ± 2.7 in wild-type embryos, mean ± SEM; P<0.01) and length (3.9 ± 0.2 μm in morphant versus 6.2 ± 0.2 μm in wild-type embryos, mean ± SEM; P<0.001) of single cilia decreased significantly (Fig.5B-C). More importantly, cilia defects in yap morphants could be rescued by overexpression of full-length yap mRNA or ΔTAD yap mRNA, but not by yapS127AmRNA (Supplementary Fig.S4).


Yes-associated protein (Yap) is necessary for ciliogenesis and morphogenesis during pronephros development in zebrafish (Danio Rerio).

He L, Xu W, Jing Y, Wu M, Song S, Cao Y, Mei C - Int. J. Biol. Sci. (2015)

yap morphants have ciliary defects. (A) In the eye, the outer segment of photoreceptor cell, which is a modified cilium, is hard to detect in the morphants at 3 d.p.f (the left arrow indicates the control, the right arrow shows defect in yap morphant). Bar: 25 μm (B) Compared with the injection control, cilia defects are observed in medial and posterior segment of the pronephric duct in the morphant. Bar: 10 μm. (C) Statistical analysis of the single cilia number per high power field and single cilia length in control and morphant embryos. The cilia are shown by staining with antibody against acetylated α-tublin. α-tub, anti-acetylated α-tublin; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; MD, medial segment of pronephric duct; PD, posterior segment f pronephric duct; MC, multicilia; SC, single cilia; HPF, high power field.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4495411&req=5

Figure 5: yap morphants have ciliary defects. (A) In the eye, the outer segment of photoreceptor cell, which is a modified cilium, is hard to detect in the morphants at 3 d.p.f (the left arrow indicates the control, the right arrow shows defect in yap morphant). Bar: 25 μm (B) Compared with the injection control, cilia defects are observed in medial and posterior segment of the pronephric duct in the morphant. Bar: 10 μm. (C) Statistical analysis of the single cilia number per high power field and single cilia length in control and morphant embryos. The cilia are shown by staining with antibody against acetylated α-tublin. α-tub, anti-acetylated α-tublin; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; MD, medial segment of pronephric duct; PD, posterior segment f pronephric duct; MC, multicilia; SC, single cilia; HPF, high power field.
Mentions: Defects in ciliary genes often lead to pronephric cyst, curved tails, heart edema, smaller eyes in zebrafish as yap morphants exhibit. To examine the effect of Yap in ciliogenesis, we stained the eyes and pronephric ducts of yap morphants and control embryos with acetylated α-tublin. In zebrafish, the outer segment of the photoreceptor is a highly modified cilium giving access to visual transduction, it begins to form by 54 h.p.f and persists thereafter. We found that the outer segments were almost discernable in smaller eyes of 3 d.p.f yap morphants (Fig.5A; arrows). The pronephric duct features multiciliated cells (MCCs) and single ciliated cells (SCCs), which form clustered cilia bundles and single cilia respectively. The single cilia are observed as early as 20 h.p.f while the bundling cilia can only be detected from about 36 h.p.f. We examined cilia formation at 3 d.p.f in at least 7 embryos each group, and found that the MCCs in the mid region of the duct did not form normal bundles but only disordered cilia, which had also been seen in other zebrafish cilia mutants 9, 39, while in the posterior segments, the number (20.3 ± 1.5 in morphant versus 30.3 ± 2.7 in wild-type embryos, mean ± SEM; P<0.01) and length (3.9 ± 0.2 μm in morphant versus 6.2 ± 0.2 μm in wild-type embryos, mean ± SEM; P<0.001) of single cilia decreased significantly (Fig.5B-C). More importantly, cilia defects in yap morphants could be rescued by overexpression of full-length yap mRNA or ΔTAD yap mRNA, but not by yapS127AmRNA (Supplementary Fig.S4).

Bottom Line: We examined expression of Yap during zebrafish embryogenesis, and its transcripts were detected in pronephric duct, while Yap protein was found to be localized in the cytoplasm and apical membrane in kidney epithelium cells.With transgenic Tg(Na(+)/K(+) ATPase:EGFP), we found that lacking Yap led to expansion and discontinuities of pronephric duct, as well as disorganization of cloaca during pronephros morphogenesis.Taken together, our data reveals that Yap is required for pronephric duct integrity, maintenance of baso-lateral cell polarity, and ciliogenesis during zebrafish kidney development.

View Article: PubMed Central - PubMed

Affiliation: 1. Kidney Institute of CPLA, Division of Nephrology, Changzheng Hospital, Second Military Medical University, NO.415 Fengyang Road, Shanghai 200003, China.

ABSTRACT
The Hippo signaling pathway and its transcriptional co-activator Yap are known as essential regulators for cell proliferation and organ size. However, little is known about their roles in kidney development and ciliogenesis. We examined expression of Yap during zebrafish embryogenesis, and its transcripts were detected in pronephric duct, while Yap protein was found to be localized in the cytoplasm and apical membrane in kidney epithelium cells. By morpholino (MO) knockdown of yap expression in zebrafish, the injected larve exhibits pronephic cysts and many aspects of ciliopathy, which can be rescued by full-length yap mRNA, but not yap (S127A) mRNA. With transgenic Tg(Na(+)/K(+) ATPase:EGFP), we found that lacking Yap led to expansion and discontinuities of pronephric duct, as well as disorganization of cloaca during pronephros morphogenesis. Mis-located Na(+)/K(+) ATPase and ciliary abnormalities are also detected in pronephric duct of yap morphants. In addition, genetic analysis suggests that yap interacts with ift20, ift88 and arl13b in pronephric cyst formation. Taken together, our data reveals that Yap is required for pronephric duct integrity, maintenance of baso-lateral cell polarity, and ciliogenesis during zebrafish kidney development.

No MeSH data available.


Related in: MedlinePlus