A specific isoform of Pyd/ZO-1 mediates junctional remodeling and formation of slit diaphragms

Carrasco‐Rando, Marta; Prieto-Sánchez, Silvia; Culí, Joaquim; Tutor, Antonio S.; Ruiz‐Gómez, Mar

doi:10.1083/jcb.201810171

Cited by 25 publications

(34 citation statements)

References 48 publications

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“…Indeed, several proteins of the adherens-and tight junctions can also be found to be components of the slit diaphragm, e.g. ZO-1, Crumbs, PAR/aPKC-complex (Huber et al, 2003b;Hartleben et al, 2008;Hirose et al, 2009;Weavers et al, 2009;Hartleben et al, 2013;Itoh et al, 2014;Satoh et al, 2014;Hochapfel et al, 2017;Weide et al, 2017;Carrasco-Rando et al, 2019;Moller-Kerutt et al, 2021). Thus, it is likely that changes in PI(4,5)P2 and PI(3,4,5)P3 affect slit diaphragm formation and maintenance/stability like they affect adherens junctions/tight junctions in classical epithelia.…”

Section: Discussionmentioning

confidence: 99%

“…Apart from the core components of the Nephrin/Neph1 family, the slit diaphragm is stabilized by adapter proteins, e.g. the Podocin homologue Mec2 (Weavers et al, 2009;Zhang et al, 2013b) and the ZO-1 homologue Polychaetoid (Weavers et al, 2009;Carrasco-Rando et al, 2019). Furthermore, we recently showed, that regulators of classical apical-basal polarity in epithelia are partly localized to slit diaphragm complexes (Heiden et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

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PI(4,5)P2 controls slit diaphragm formation and endocytosis in Drosophila nephrocytes

Gass

Borkowsky

Lotz

et al. 2021

Preprint

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Drosophila nephrocytes are an emerging model system for mammalian podocytes and podocyte-associated diseases. Like podocytes, nephrocytes exhibit characteristics of epithelial cells, but the role of phospholipids in polarization of these cells is yet unclear. In epithelia phosphatidylinositol(4,5)bisphosphate (PI(4,5)P2) and phosphatidylinositol(3,4,5)-trisphosphate (PI(3,4,5)P3) are asymmetrically distributed in the plasma membrane and determine apical-basal polarity. Here we demonstrate that both phospholipids are present in the plasma membrane of nephrocytes, but only PI(4,5)P2 accumulates at slit diaphragms. Knockdown of Skittles, a phosphatidylinositol(4)phosphate 5-kinase, which produces PI(4,5)P2, abolished slit diaphragm formation and led to strongly reduced endocytosis. Notably, reduction in PI(3,4,5)P3 by overexpression of PTEN or expression of a dominant-negative phosphatidylinositol-3-Kinase did not affect nephrocyte function, whereas enhanced formation of PI(3,4,5)P3 by constitutively active phosphatidylinositol-3-Kinase resulted in strong slit diaphragm and endocytosis defects by ectopic activation of the Akt/mTOR pathway. Thus, PI(4,5)P2 but not PI(3,4,5)P3 is essential for slit diaphragm formation and nephrocyte function. However, PI(3,4,5)P3 has to be tightly controlled to ensure nephrocyte development.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

PI(4,5)P2 controls slit diaphragm formation and endocytosis in Drosophila nephrocytes

Gass

Borkowsky

Lotz

et al. 2021

Preprint

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“…Guinea-pig anti-Duf extracellular (1:100), rabbit anti-Duf extracellular (1:400) (Weavers et al, 2009), rabbit anti-Pyd (1:100), rat anti-PydEx5 (1:200) (Carrasco-Rando et al, 2019), rabbit anti-GFP (1:300) (ThermoFisher, Cat # A-6455), rabbit Cy TM 3-anti-HRP (1:100) (Jackson ImmunoResearch, Cat# AB_2340262), mouse anti-V5 (1:500) (ThermoFisher, Cat # R960-25), mouse anti-cMYC 9E-10 (1:50 from DSHB; 1:1000 from BAbCO Cat# 1244636002), rat anti-HA High Affinity (1:500) (Merk ROAHAHA Roche, Cat# 11867423001), guinea-pig anti-Boca (1:200) (Culi and Mann, 2003), guinea-pig anti-Sec15 (1:100) (Mehta et al, 2005), guinea-pig anti-dAMN (1.100) (Ivy et al, 2015), guinea-pig anti-Cubn-2 (1:200), rat anti-Cubn (1:400), guinea-pig anti-Mgl (1:500) (this study). (http://bacpacresources.org//), hAMN cDNA clone (Pedersen et al, 2010).…”

Section: Antibodiesmentioning

confidence: 99%

“…The seminal discovery that Drosophila nephrocytes, the excretory cells engaged in haemolymph ultrafiltration, had filtration diaphragms analogous to the SD of vertebrate podocytes (Weavers et al, 2009;Zhuang et al, 2009) opened the door to many studies that used them to examine different aspects of SD formation and stability (Carrasco-Rando et al, 2019;Tutor et al, 2014). Moreover, Drosophila nephrocytes also helped investigate the effects on podocyte function of specific mutations associated to steroid-resistant nephrotic syndromes by generating the corresponding fly avatars (Gonçalves et al, 2018;Lovric et al, 2017;Zhao et al, 2019;Zhu et al, 2017) and to model nephropathies (Hermle et al, 2016;Na et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Endocytosis mediated by an atypical CUBAM complex modulates slit diaphragm dynamics in nephrocytes

et al. 2021

Self Cite

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The vertebrate endocytic receptor CUBAM, consisting of three cubilin monomers complexed with a single amnionless molecule, plays a major role in protein reabsorption in the renal proximal tubule. Here, we show that Drosophila CUBAM is a tripartite complex composed of Amnionless and two cubilin paralogues, Cubilin and Cubilin2, and that it is required for nephrocyte slit diaphragm (SD) dynamics. Loss of CUBAM-mediated endocytosis induces dramatic morphological changes in nephrocytes and promotes enlarged ingressions of the external membrane and SD mislocalisation. These phenotypes result in part from an imbalance between endocytosis, which is strongly impaired in CUBAM mutants, and exocytosis in these highly active cells. Of note, rescuing receptor-mediated endocytosis by Megalin/LRP2 or Rab5 expression only partially restores SD positioning in CUBAM mutants, suggesting a specific requirement of CUBAM in SD degradation and/or recycling. This finding and the reported expression of CUBAM in podocytes suggest a possible unexpected conserved role for this endocytic receptor in vertebrate SD remodelling.

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“…For podocytes it has already been shown, that protein complexes, which are essential for TJ assembly and -function, are also important regulators of slit diaphragm assembly/maintenance. One example is the adaptor protein Zonula Occludens protein 1 [22,23], whose Drosophila homologue Polychaetoid (Pyd) has recently been confirmed to regulate slit diaphragm assembly in nephrocytes, too [24]. PARtitioning defective 3 (PAR-3) and atypical Protein Kinase C (aPKC) both control TJ formation and are also involved in slit diaphragm assembly [25][26][27][28][29][30].…”

Section: Introductionmentioning

confidence: 99%

Apical-basal polarity regulators are essential for slit diaphragm assembly and endocytosis in Drosophila nephrocytes

Heiden

Siwek

Lotz

et al. 2021

Cell. Mol. Life Sci.

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Apical-basal polarity is a key feature of most epithelial cells and it is regulated by highly conserved protein complexes. In mammalian podocytes, which emerge from columnar epithelial cells, this polarity is preserved and the tight junctions are converted to the slit diaphragms, establishing the filtration barrier. In Drosophila, nephrocytes show several structural and functional similarities with mammalian podocytes and proximal tubular cells. However, in contrast to podocytes, little is known about the role of apical-basal polarity regulators in these cells. In this study, we used expansion microscopy and found the apical polarity determinants of the PAR/aPKC and Crb-complexes to be predominantly targeted to the cell cortex in proximity to the nephrocyte diaphragm, whereas basolateral regulators also accumulate intracellularly. Knockdown of PAR-complex proteins results in severe endocytosis and nephrocyte diaphragm defects, which is due to impaired aPKC recruitment to the plasma membrane. Similar, downregulation of most basolateral polarity regulators disrupts Nephrin localization but had surprisingly divergent effects on endocytosis. Our findings suggest that morphology and slit diaphragm assembly/maintenance of nephrocytes is regulated by classical apical-basal polarity regulators, which have distinct functions in endocytosis.

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A specific isoform of Pyd/ZO-1 mediates junctional remodeling and formation of slit diaphragms

Abstract: Podocyte slit diaphragms are key components of the glomerular filtration barrier. Using Drosophila nephrocytes, Carrasco-Rando et al. propose a conserved role for Pyd/ZO-1 in triggering junctional remodeling leading to the formation of slit diaphragms.

Cited by 25 publications

References 48 publications

PI(4,5)P2 controls slit diaphragm formation and endocytosis in Drosophila nephrocytes

PI(4,5)P2 controls slit diaphragm formation and endocytosis in Drosophila nephrocytes

Endocytosis mediated by an atypical CUBAM complex modulates slit diaphragm dynamics in nephrocytes

Apical-basal polarity regulators are essential for slit diaphragm assembly and endocytosis in Drosophila nephrocytes

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