The fast-recycling receptor Megalin defines the apical recycling pathway of epithelial cells

Bay, Andrés E. Perez; Schreiner, Ryan; Benedicto, Ignacio; Marzolo, Marı́a Paz; Banfelder, Jason R.; Weinstein, Alan M.; Rodríguez-Boulan, Enrique

doi:10.1038/ncomms11550

Cited by 76 publications

(90 citation statements)

References 61 publications

(124 reference statements)

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“…In polarized epithelial cells, Rab11a is also known to mediate vesicle trafficking and recycling ( Perez Bay et al, 2016 ; Weisz and Rodriguez-Boulan, 2009 ). At this time, it is unclear if corin undergoes endocytosis and recycling.…”

Section: Discussionmentioning

confidence: 99%

A conserved LDL-receptor motif regulates corin and CD320 membrane targeting in polarized renal epithelial cells

Zhang

Chen

Sun

et al. 2020

eLife

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Selective protein distribution on distinct plasma membranes is important for epithelial cell function. To date, how proteins are directed to specific epithelial cell surface is not fully understood. Here we report a conserved DSSDE motif in LDL-receptor (LDLR) modules of corin (a transmembrane serine protease) and CD320 (a receptor for vitamin B12 uptake), which regulates apical membrane targeting in renal epithelial cells. Altering this motif prevents specific apical corin and CD320 expression in polarized Madin–Darby canine kidney (MDCK) cells. Mechanistic studies indicate that this DSSDE motif participates in a Rab11a-dependent mechanism that specifies apical sorting. In MDCK cells, inhibition of Rab11a, but not Rab11b, expression leads to corin and CD320 expression on both apical and basolateral membranes. Together, our results reveal a novel molecular recognition mechanism that regulates LDLR module-containing proteins in their specific apical expression in polarized renal epithelial cells.

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Section: Discussionmentioning

confidence: 99%

A conserved LDL-receptor motif regulates corin and CD320 membrane targeting in polarized renal epithelial cells

Zhang

Chen

Sun

et al. 2020

eLife

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“…Reduction of LRP2 expression was attained in mice, but a concomitant protein expression decrease was not observed, even after serial administration. A number of factors may contribute to this finding, including (1) the high proportion of membrane-bound protein as compared to cytoplasmic protein, (2) the modest degree of mRNA reduction, (3) the rapid rebound and increase in mRNA expression following siRNA delivery, and (4) the relatively long half-life of the protein (4.8 h) [ 25 ]. These results support the theoretical use of siRNA to transiently reduce baseline expression of transcripts in the kidney; however, the results also suggest that the practical application of this technique will prove difficult.…”

Section: Discussionmentioning

confidence: 99%

“…While this may appear counterintuitive to knock down the very protein responsible uptake, in this scenario, it is unlikely to have had a remarkable effect since (1) megalin's uptake capacity is high, and (2) protein expression reduction was never achieved. A second factor that may have affected efficacy is the rapid and repetitive recycling of the protein [ 25 ]. Megalin recycles from the cytoplasm to the cell membrane every 1.2 minutes.…”

Section: Discussionmentioning

confidence: 99%

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In Vivo siRNA Delivery and Rebound of Renal LRP2 in Mice

Eadon

Cheng

Hato

et al. 2017

Journal of Drug Delivery

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siRNA stabilized for in vivo applications is filtered and reabsorbed in the renal proximal tubule (PT), reducing mRNA expression transiently. Prior siRNA efforts have successfully prevented upregulation of mRNA in response to injury. We proposed reducing constitutive gene and protein expression of LRP2 (megalin) in order to understand its molecular regulation in mice. Using siRNA targeting mouse LRP2 (siLRP2), reduction of LRP2 mRNA expression was compared to scrambled siRNA (siSCR) in mouse PT cells. Mice received siLRP2 administration optimized for dose, administration site, carrier solution, administration frequency, and administration duration. Kidney cortex was collected upon sacrifice. Renal gene and protein expression were compared by qRT-PCR, immunoblot, and immunohistochemistry (IHC). Compared to siSCR, siLRP2 reduced mRNA expression in PT cells to 16.6% ± 0.6%. In mouse kidney cortex, siLRP2 reduced mRNA expression to 74.8 ± 6.3% 3 h and 70.1 ± 6.3% 6 h after administration. mRNA expression rebounded at 12 h (160.6 ± 11.2%). No megalin renal protein expression reduction was observed by immunoblot or IHC, even after serial twice daily dosing for 3.5 days. Megalin is a constitutively expressed protein. Although LRP2 renal mRNA expression reduction was achieved, siRNA remains a costly and inefficient intervention to reduce in vivo megalin protein expression.

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“…Megalin, or LDL receptor-related protein 2 (LRP2), is a member of the LDL receptor (LDLR) family and is an endocytic receptor expressed on the apical surface of several epithelial cells that internalizes a variety of ligands [ 15 ]. Megalin is considered to define the apical recycling pathway of epithelial cells and to have an influence on the high-protein absorption capacity of the kidney proximal tubule, and on genetic proteinuric syndromes [ 16 ]. Megalin is involved in the uptake of drugs and toxic substances such as denatured protein, and also in the renal tubule cell disorder by the excessive load of plasma protein [ 17–19 ].…”

Section: Introductionmentioning

confidence: 99%

The effect of cholesterol overload on mouse kidney and kidney-derived cells

et al. 2018

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Introduction: Dyslipidemia is one of the onset and risk factors of chronic kidney disease and renal function drop is seen in lipoprotein abnormal animal models. However, the detailed molecular mechanism of renal lipotoxicity has not been clarified. Therefore, the present study aimed to investigate the influence of cholesterol overload using mouse kidney tissue and kidney-derived cultured cells.Methods: C57BL/6 mice were fed normal diet (ND) or 1.25% cholesterol-containing high-cholesterol diet (HCD) for 11 weeks, and we used megalin as a proximal tubule marker for immunohistology. We added beta-very low density lipoprotein (βVLDL) to kidney-derived cells and examined the effect of cholesterol overload on megalin protein and mRNA expression level, cell proliferation and cholesterol content in cells.Results: In the kidney of HCD mice, the gap between glomerulus and the surrounding Bowman’s capsule decreased and the expression level of megalin decreased. After βVLDL treatment to the cells, the protein expression and mRNA expression level of megalin decreased and cell proliferation was restrained. We also observed an increase in cholesterol accumulation in the cell and free cholesterol/phospholipid ratios increased.Conclusions: These findings suggest that the increased cholesterol load on kidney contribute to the decrease of megalin and the overloaded cholesterol is taken into the renal tubule epithelial cells, causing suppression on cell proliferation, which may be the cause of kidney damage.

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The fast-recycling receptor Megalin defines the apical recycling pathway of epithelial cells

Cited by 76 publications

References 61 publications

A conserved LDL-receptor motif regulates corin and CD320 membrane targeting in polarized renal epithelial cells

A conserved LDL-receptor motif regulates corin and CD320 membrane targeting in polarized renal epithelial cells

In Vivo siRNA Delivery and Rebound of Renal LRP2 in Mice

The effect of cholesterol overload on mouse kidney and kidney-derived cells

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