Exocytosis of Vacuolar Apical Compartment (VAC) in Madin-Darby Canine Kidney Epithelial Cells: cAMP Is Involved as Second Messenger

Brignoni, Mirtha; Podestá, Ernesto J.; Mele, Pablo G.; Rodriguez, Marcelo L.; Vega-Salas, Dora E.; Salas, Pedro J.

doi:10.1006/excr.1993.1072

Cited by 18 publications

(12 citation statements)

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“…Fourth, we observed VACs and syntaxin 4 -positive basolateral organelles in MDCK cells that are grown either in regular-calcium medium for brief periods (Figure 3) or in single cells or cells at the edge of cell patches after sparse seeding and growth for 16 h (Figure 8). The VAC has been described and characterized previously in nonpolarized MDCK cells and other epithelial cell lines as well as in carcinomas (Vega-Salas et al, 1987Gilbert and Rodriguez-Boulan, 1991;Brignoni et al, 1993). In contrast, to our knowledge, the basolateral compartment that we identified here is a novel organelle that has not been described previously, perhaps because of the lack of availability of a marker protein such as syntaxin 4.…”

Section: Sh Low Et Almentioning

confidence: 52%

“…The finding that neither the apical nor the basolateral vacuole appears to be a degradative, lysosomal compartment indicates that proteins may be stored in them for later use once cell contacts have been reestablished. This is supported by the observation that VACs can be rapidly exocytosed as a whole from MDCK cells after reestablishment of cell-cell contacts (Vega-Salas et al, 1988) or after increasing the intracellular cAMP concentration (Brignoni et al, 1993).…”

Section: Sh Low Et Almentioning

confidence: 74%

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Intracellular Redirection of Plasma Membrane Trafficking after Loss of Epithelial Cell Polarity

Low

Miura²,

Roche

et al. 2000

MBoC

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In polarized Madin-Darby canine kidney epithelial cells, components of the plasma membrane fusion machinery, the t-SNAREs syntaxin 2, 3, and 4 and SNAP-23, are differentially localized at the apical and/or basolateral plasma membrane domains. Here we identify syntaxin 11 as a novel apical and basolateral plasma membrane t-SNARE. Surprisingly, all of these t-SNAREs redistribute to intracellular locations when Madin-Darby canine kidney cells lose their cellular polarity. Apical SNAREs relocalize to the previously characterized vacuolar apical compartment, whereas basolateral SNAREs redistribute to a novel organelle that appears to be the basolateral equivalent of the vacuolar apical compartment. Both intracellular plasma membrane compartments have an associated prominent actin cytoskeleton and receive membrane traffic from cognate apical or basolateral pathways, respectively. These findings demonstrate a fundamental shift in plasma membrane traffic toward intracellular compartments while protein sorting is preserved when epithelial cells lose their cell polarity. INTRODUCTIONTraffic between membranous compartments is mediated by the soluble N-ethylmaleimide-sensitive factor attachment protein (SNARE) machinery in virtually all membrane traffic pathways investigated so far (Rothman and Warren, 1994;Hanson et al., 1997;Hay and Scheller, 1997;Nichols and Pelham, 1998). During vesicle docking, membrane proteins on the vesicle membrane (v-SNAREs) and the target membrane (t-SNAREs) bind to each other to form a complex that ultimately leads to fusion of the lipid bilayers. One aspect of the SNARE hypothesis is that successful membrane fusion requires the binding of matching combinations of v-and t-SNAREs, thereby ensuring the necessary specificity of vesicle fusion. Accordingly, each membrane organelle and each class of transport vesicles should be defined by a certain set of t-and v-SNARE isoforms. Many SNAREs have been identified to date, and protein sequence analysis has shown that v-and t-SNAREs of the currently known SNARE subfamilies are evolutionarily related to each other and belong to a common superfamily (Weimbs et al., 1997b(Weimbs et al., , 1998. It is conceivable that the specificity of vesicle fusion is not directly determined by t-SNARE/v-SNARE interactions per se but rather by interactions involving larger complexes, including SNAREs and their regulatory proteins, such as those of the rab and sec1 protein families (Christoforidis et al., 1999;Gonzalez and Scheller, 1999;Pfeffer, 1999).Epithelial cells display an additional layer of complexity in that they are typically polarized and possess two distinct plasma membrane domains (Louvard et al., 1992;Simons et al., 1992;Le Gall et al., 1995;Drubin and Nelson, 1996;Yeaman et al., 1999). The apical and basolateral plasma membrane domains have different protein and lipid compositions that reflect the different functions of these domains. This plasma membrane polarity is established and maintained by protein sorting and specific vesicle trafficking routes in th...

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Section: Sh Low Et Almentioning

confidence: 52%

Section: Sh Low Et Almentioning

confidence: 74%

Intracellular Redirection of Plasma Membrane Trafficking after Loss of Epithelial Cell Polarity

Low

Miura²,

Roche

et al. 2000

MBoC

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“…In Caco-2 cysts, Cdc42 knockdown correlates with disrupted spindle orientation, without notable apical transport defects29. Caco-2 lumen formation requires artificially increasing cAMP, which strongly promotes apical exocytosis34, potentially masking detection of apical transport defects. As Rab8/11, exocyst, and Par3/aPKC also have roles in spindle orientation and/or cytokinesis35–37, the extent to which their function in cell division is separate from apical trafficking is unknown.…”

Section: Discussionmentioning

confidence: 99%

A molecular network for de novo generation of the apical surface and lumen

et al. 2010

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To form epithelial organs cells must polarize and generate de novo an apical domain and lumen. Epithelial polarization is masterminded by polarity complexes, which are thought to direct downstream events such as polarized membrane traffic, though this interconnection is not well understood. We report that Rab11a regulates apical traffic and lumen formation via the Rab GEF Rabin8, and its target Rab8a. Rab8a/11a act via the exocyst to target Par3 to the apical surface, and control apical Cdc42 activation via the Cdc42 GEF, Tuba. These components assemble at a transient apical membrane initiation site to form the lumen. This Rab11a-directed network directs Cdc42-dependent apical exocytosis during lumen formation, revealing a novel interplay of the machineries of vesicular transport and polarization.Most internal epithelial organs consist of a monolayer of polarized epithelial cells surrounding a central lumen. Polarization requires the interaction of the signaling complexes and scaffolds that define cortical domains with the polarized membrane sorting machinery 1 . In yeast, traffic from the trans-Golgi network to the surface is regulated by Ypt32p and Sec4p 2 , homologs of mammalian Rab11 and Rab8, respectively. These are linked by Sec2p (homolog of mammalian Rabin8), a guanine nucleotide exchange factor (GEF) for Sec4p, which is recruited by Ypt32p. Sec2p and Sec4p in turn interact with the exocyst, which docks vesicles to the surface 3 .Definition of cortical domains in metazoa involves a complex of Par3, Par6, atypical PKC (aPKC), and the GTPase Cdc42 4 . This complex is a master regulator of polarity, conventionally depicted upstream of membrane trafficking machinery. How this complex interfaces with membrane transport is poorly understood.Here we show a molecular mechanism for lumen and apical surface formation, linking Rab8a/ 11a, exocyst, annexin2, Cdc42 and its GEF Tuba, and the Par3/aPKC complex. This novel NIH Public Access Author ManuscriptNat Cell Biol. Author manuscript; available in PMC 2010 November 8. Published in final edited form as:Nat Cell Biol. 2010 November ; 12(11): 1035-1045. doi:10.1038/ncb2106. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript pathway shows how the membrane traffic and cortical polarity machineries cooperate to generate the apical surface and lumen. RESULTS Apical polarization during lumen formationUpon plating into 3D culture, individual MDCK cells proliferate and assemble into cyst structures -a polarized spherical monolayer surrounding a central lumen. Lumenogenesis requires the apical membrane determinant gp135/podocalyxin 5 (PCX in figures). Initially, MDCK aggregates have podocalyxin at the ECM-contacting surface (Fig. 1a, 12 h; Fig. S1a), before polarity inversion occurs, with β-catenin and Na/K-ATPase at cell-cell junctions and podocalyxin now at the lumen (Fig. 1a, 24-48 h, arrows; Fig. S1d) 6,7 . Early lumens occur at a site previously termed the "Pre-Apical Patch" (PAP), where opposing plasma membranes are separated, but the po...

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“…A major difference between non-polarized cells of epithelial and non-epithelial origin may therefore be that in the former plasma membrane and secretory proteins are retained inside the cell rather than delivered to the surface. The vacuolar apical compartment has been described and characterized previously in nonpolarized MDCK cells and other epithelial cell lines as well as in a variety of carcinomas (3,11,(38)(39)(40). In contrast, to our knowledge, the basolateral compartment that we identified here is a novel organelle that has not been described previously, perhaps because of the lack of availability of a marker protein such as syntaxin 4.…”

Section: Bodymentioning

confidence: 50%

“…The finding that neither the apical nor basolateral vacuoles appear to be degradative, lysosomal compartments indicates that proteins may be stored in them for later use once cell contacts have been re-established. This is supported by the observation that VACs can be rapidly exocytosed as a whole from MDCK cells after re-establishment of cell-cell contacts (39) or after raising the intracellular cAMP concentration (3). A basolateral equivalent of the VAC may be used to temporarily store proteins such as integrins.…”

Section: Bodymentioning

confidence: 88%

Effects of c-Myc and TGF-Alpha on Polarized Membrane Traffic

Altschuler¹,

Mostov²

2000

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1 069u r REPORT DOCUMENTATION PAGE I 0MBNo. Public reporting burde~nfor this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Washington Headquarters Services, Directorate for Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302, and to the Office of Management and Budget, Paperwork Reduction Project (0704-0188), Washington, DC 20503

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Exocytosis of Vacuolar Apical Compartment (VAC) in Madin-Darby Canine Kidney Epithelial Cells: cAMP Is Involved as Second Messenger

Cited by 18 publications

References 0 publications

Intracellular Redirection of Plasma Membrane Trafficking after Loss of Epithelial Cell Polarity

Intracellular Redirection of Plasma Membrane Trafficking after Loss of Epithelial Cell Polarity

A molecular network for de novo generation of the apical surface and lumen

Effects of c-Myc and TGF-Alpha on Polarized Membrane Traffic

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