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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.

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Related in: MedlinePlus

The Na+/K+ ATPase is mis-located to the apical surface of pronephric duct in yap morphants. (A-B) Whole-mount staining shows that the inner diameter of the enlarged duct indicated by aPKC is about fourfold of the control, however, the change of the outer diameter indicated with α-6F is not that dramatic. (C-D') The cross section view demonstrates increased cell number surrounding the dilated duct compared with the control duct (white dashed circles represent the cross sections of the ducts), the baso-lateral staining of Na+/K+ ATPase is not obvious in a part of enlarged tubules (Fig.4D'; red arrow), and even is mis-targeted to the apical surface adjacent to aPKC (D'; arrowhead). (C' and D') Arrows indicate basal surface and asterisks indicate lateral surface. (A-B) Bar: 10 μm. (C-D) Bar: 5 μm.
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Figure 4: The Na+/K+ ATPase is mis-located to the apical surface of pronephric duct in yap morphants. (A-B) Whole-mount staining shows that the inner diameter of the enlarged duct indicated by aPKC is about fourfold of the control, however, the change of the outer diameter indicated with α-6F is not that dramatic. (C-D') The cross section view demonstrates increased cell number surrounding the dilated duct compared with the control duct (white dashed circles represent the cross sections of the ducts), the baso-lateral staining of Na+/K+ ATPase is not obvious in a part of enlarged tubules (Fig.4D'; red arrow), and even is mis-targeted to the apical surface adjacent to aPKC (D'; arrowhead). (C' and D') Arrows indicate basal surface and asterisks indicate lateral surface. (A-B) Bar: 10 μm. (C-D) Bar: 5 μm.

Mentions: In yap morphants, cystic dilation in the glomeruli-neck region were first detected under light microscopy at about 50-55 h.p.f. Pronephric cyst is sometimes accompanied by tubular dilation, so we further examined pronephric duct with immunostaining. aPKC indicates the apical membrane as inner diameter of the pronephric duct, and α6F subunit of Na+/K+ ATPase represents basolateral surface as outer diameter. We found tubular dilation develop in both proximal and distal segments of pronephros in 3 d.p.f morphants, and the proximal tubule remained straight with no sign of “hairpin” convolution (Fig.4A-B and S2A). Some studies have reported that altered apico-basal cell polarity in renal epithelia may lead to tubular expansion and cyst formation, e.g. mis-sorting or insertion of transporters and receptors as Na+/K+ ATPase, NKCC1 and EGFR38. Thus we analyzed Na+/K+ ATPase localization in 3 d.p.f embryos with cross section staining. The baso-lateral staining of Na+/K+ ATPase was not obvious in a part of enlarged tubules (Fig.4D'; red arrow), and even apical expression was detected (Fig.4D'; arrowhead). Mis-localization of Na+/K+ ATPase in renal cells can result in secretion of sodium into the lumen and fluid accumulation39.Meanwhile, location of the apical marker, such as aPKC and F-actin, appeared intact (Fig. 4D and S2B). The results suggest that disruption of baso-lateral cell polarity, but not apical polarity, may give rise to cyst formation in yap morphants.


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)

The Na+/K+ ATPase is mis-located to the apical surface of pronephric duct in yap morphants. (A-B) Whole-mount staining shows that the inner diameter of the enlarged duct indicated by aPKC is about fourfold of the control, however, the change of the outer diameter indicated with α-6F is not that dramatic. (C-D') The cross section view demonstrates increased cell number surrounding the dilated duct compared with the control duct (white dashed circles represent the cross sections of the ducts), the baso-lateral staining of Na+/K+ ATPase is not obvious in a part of enlarged tubules (Fig.4D'; red arrow), and even is mis-targeted to the apical surface adjacent to aPKC (D'; arrowhead). (C' and D') Arrows indicate basal surface and asterisks indicate lateral surface. (A-B) Bar: 10 μm. (C-D) Bar: 5 μm.
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Related In: Results  -  Collection

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Figure 4: The Na+/K+ ATPase is mis-located to the apical surface of pronephric duct in yap morphants. (A-B) Whole-mount staining shows that the inner diameter of the enlarged duct indicated by aPKC is about fourfold of the control, however, the change of the outer diameter indicated with α-6F is not that dramatic. (C-D') The cross section view demonstrates increased cell number surrounding the dilated duct compared with the control duct (white dashed circles represent the cross sections of the ducts), the baso-lateral staining of Na+/K+ ATPase is not obvious in a part of enlarged tubules (Fig.4D'; red arrow), and even is mis-targeted to the apical surface adjacent to aPKC (D'; arrowhead). (C' and D') Arrows indicate basal surface and asterisks indicate lateral surface. (A-B) Bar: 10 μm. (C-D) Bar: 5 μm.
Mentions: In yap morphants, cystic dilation in the glomeruli-neck region were first detected under light microscopy at about 50-55 h.p.f. Pronephric cyst is sometimes accompanied by tubular dilation, so we further examined pronephric duct with immunostaining. aPKC indicates the apical membrane as inner diameter of the pronephric duct, and α6F subunit of Na+/K+ ATPase represents basolateral surface as outer diameter. We found tubular dilation develop in both proximal and distal segments of pronephros in 3 d.p.f morphants, and the proximal tubule remained straight with no sign of “hairpin” convolution (Fig.4A-B and S2A). Some studies have reported that altered apico-basal cell polarity in renal epithelia may lead to tubular expansion and cyst formation, e.g. mis-sorting or insertion of transporters and receptors as Na+/K+ ATPase, NKCC1 and EGFR38. Thus we analyzed Na+/K+ ATPase localization in 3 d.p.f embryos with cross section staining. The baso-lateral staining of Na+/K+ ATPase was not obvious in a part of enlarged tubules (Fig.4D'; red arrow), and even apical expression was detected (Fig.4D'; arrowhead). Mis-localization of Na+/K+ ATPase in renal cells can result in secretion of sodium into the lumen and fluid accumulation39.Meanwhile, location of the apical marker, such as aPKC and F-actin, appeared intact (Fig. 4D and S2B). The results suggest that disruption of baso-lateral cell polarity, but not apical polarity, may give rise to cyst formation in yap morphants.

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