A common strategy to improve transmembrane transport in polarized epithelial cells based on sorting signals: Guiding nanocarriers to TGN rather than to the basolateral plasma membrane directly

2021 ◽  
Author(s):  
Runyu Zhang ◽  
Hailiang Deng ◽  
Yuxing Lin ◽  
Xing Wang ◽  
Bing He ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Transmembrane Transport ◽  
Sorting Signals ◽  
Common Strategy ◽  
Basolateral Plasma Membrane ◽  
Polarized Epithelial Cells

scholarly journals Chlorpromazine Induces Basolateral Aquaporin-2 Accumulation via F-Actin Depolymerization and Blockade of Endocytosis in Renal Epithelial Cells

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 1057
Author(s):  
Richard Bouley ◽  
Naofumi Yui ◽  
Abby Terlouw ◽  
Pui W. Cheung ◽  
Dennis Brown
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Mdck Cells ◽  
Rat Kidney ◽  
Apical Plasma Membrane ◽  
Rhodamine Phalloidin ◽  
Renal Epithelial Cells ◽  
Aquaporin 2 ◽  
Basolateral Plasma Membrane

We previously showed that in polarized Madin–Darby canine kidney (MDCK) cells, aquaporin-2 (AQP2) is continuously targeted to the basolateral plasma membrane from which it is rapidly retrieved by clathrin-mediated endocytosis. It then undertakes microtubule-dependent transcytosis toward the apical plasma membrane. In this study, we found that treatment with chlorpromazine (CPZ, an inhibitor of clathrin-mediated endocytosis) results in AQP2 accumulation in the basolateral, but not the apical plasma membrane of epithelial cells. In MDCK cells, both AQP2 and clathrin were concentrated in the basolateral plasma membrane after CPZ treatment (100 µM for 15 min), and endocytosis was reduced. Then, using rhodamine phalloidin staining, we found that basolateral, but not apical, F-actin was selectively reduced by CPZ treatment. After incubation of rat kidney slices in situ with CPZ (200 µM for 15 min), basolateral AQP2 and clathrin were increased in principal cells, which simultaneously showed a significant decrease of basolateral compared to apical F-actin staining. These results indicate that clathrin-dependent transcytosis of AQP2 is an essential part of its trafficking pathway in renal epithelial cells and that this process can be inhibited by selectively depolymerizing the basolateral actin pool using CPZ.

scholarly journals Apical Scaffolding Protein NHERF2 Modulates the Localization of Alternatively Spliced Plasma Membrane Ca2+Pump 2B Variants in Polarized Epithelial Cells

2010 ◽  
Vol 285 (41) ◽  
pp. 31704-31712 ◽  
Author(s):  
Rita Padányi ◽  
Yuning Xiong ◽  
Géza Antalffy ◽  
Krisztina Lór ◽  
Katalin Pászty ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Scaffolding Protein ◽  
Alternatively Spliced ◽  
Polarized Epithelial Cells

scholarly journals Arf6 regulates AP-1B–dependent sorting in polarized epithelial cells

2011 ◽  
Vol 194 (6) ◽  
pp. 873-887 ◽  
Author(s):  
Elina Shteyn ◽  
Lucy Pigati ◽  
Heike Fölsch
Keyword(s):  
Epithelial Cells ◽  
Dominant Negative ◽  
Recycling Endosomes ◽  
Membrane Recruitment ◽  
Important Regulator ◽  
Basolateral Plasma Membrane ◽  
Polarized Epithelial Cells ◽  
Clathrin Adaptor ◽  
Basolateral Targeting

The epithelial cell–specific clathrin adaptor complex AP-1B facilitates the sorting of various transmembrane proteins from recycling endosomes (REs) to the basolateral plasma membrane. Despite AP-1B’s clear importance in polarized epithelial cells, we still do not fully understand how AP-1B orchestrates basolateral targeting. Here we identify the ADP-ribosylation factor 6 (Arf6) as an important regulator of AP-1B. We show that activated Arf6 pulled down AP-1B in vitro. Furthermore, interfering with Arf6 function through overexpression of dominant-active Arf6Q67L or dominant-negative Arf6D125N, as well as depletion of Arf6 with short hairpin RNA (shRNA), led to apical missorting of AP-1B–dependent cargos. In agreement with these data, we found that Arf6 colocalized with AP-1B and transferrin receptor (TfnR) in REs. In addition, we observed specific recruitment of AP-1B into Arf6-induced membrane ruffles in nonpolarized cells. We conclude that activated Arf6 directs membrane recruitment of AP-1B, thus regulating AP-1B’s functions in polarized epithelial cells.

scholarly journals Taking the Scenic Route: Biosynthetic Traffic to the Plasma Membrane in Polarized Epithelial Cells

Traffic ◽  
2009 ◽  
Vol 10 (8) ◽  
pp. 972-981 ◽  
Author(s):  
Heike Fölsch ◽  
Polly E. Mattila ◽  
Ora A. Weisz
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Polarized Epithelial Cells

scholarly journals Plasma membrane protein sorting in polarized epithelial cells.

1992 ◽  
Vol 116 (3) ◽  
pp. 577-583 ◽  
Author(s):  
K Mostov ◽  
G Apodaca ◽  
B Aroeti ◽  
C Okamoto
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Membrane Protein ◽  
Protein Sorting ◽  
Plasma Membrane Protein ◽  
Polarized Epithelial Cells

The ΔF508 mutation shortens the biochemical half-life of plasma membrane CFTR in polarized epithelial cells

2001 ◽  
Vol 280 (1) ◽  
pp. C166-C174 ◽  
Author(s):  
Ghanshyam D. Heda ◽  
Mridul Tanwani ◽  
Christopher R. Marino
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Half Life ◽  
Immunoblot Analysis ◽  
Wild Type ◽  
Δf508 Cftr ◽  
Polarized Epithelial Cells ◽  
Physical And Chemical ◽  
Biochemical Stability

Although the biosynthetic arrest of the ΔF508 mutant of cystic fibrosis transmembrane conductance regulator (CFTR) can be partially reversed by physical and chemical means, recent evidence suggests that the functional stability of the mutant protein after reaching the cell surface is compromised. To understand the molecular basis for this observation, the current study directly measured the half-life of ΔF508 and wild-type CFTR at the cell surface of transfected LLC-PK1 cells. Plasma membrane CFTR expression over time was characterized biochemically and functionally in these polarized epithelial cells. Surface biotinylation, streptavidin extraction, and quantitative immunoblot analysis determined the biochemical half-life of plasma membrane ΔF508 CFTR to be ∼4 h, whereas the plasma membrane half-life of wild-type CFTR exceeded 48 h. This difference in biochemical stability correlated with CFTR-mediated transport function. These findings indicate that the ΔF508 mutation decreases the biochemical stability of CFTR at the cell surface. We conclude that the ΔF508 mutation triggers more rapid internalization of CFTR and/or its preferential sorting to a pathway of rapid degradation.

Sign in / Sign up

Export Citation Format

Share Document