Abstract
We report on the characteristics of raft domains in the apical membrane from human placental syncytiotrophoblast (hSTB), an epithelium responsible for maternal–fetal exchange. Previously, we described two isolated fractions of the hSTB apical membrane: a classical microvillous membrane (MVM) and a light microvillous membrane (LMVM). Detergent-resistant microdomains (DRMs) from MVM and LMVM were prepared with Triton X-100 followed by flotation in a sucrose gradient and tested by Western and dot blot with raft markers (placental alkaline phosphatase, lipid ganglioside, annexin 2) and transferrin receptor as a nonraft marker. DRMs from both fractions showed a consistent peak for these markers, except that the DRMs from MVM had no annexin 2 mark. Cholesterol depletion modified the segregation in both groups of DRMs. Our results show two distinguishable lipid raft subsets from MVM and LMVM. Additionally, we found significant differences between MVM and LMVM in cholesterol content and in expression of cytoskeletal proteins. MVM is enriched in ezrin and β-actin; in contrast, cholesterol and cytokeratin-7 are more abundant in LMVM. These differences may explain the distinct properties of the lipid raft subtypes.
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Abbreviations
- hSTB:
-
Syncytiotrophoblast
- PLAP:
-
Alkaline phosphatase
- Anx-2:
-
Annexin A2
- MVM:
-
Classical microvillous membrane
- LMVM:
-
Light microvillous membrane
- CK-7:
-
Cytokeratin-7
- htf-R:
-
Human transferrin receptor
- DRMs:
-
Detergent-resistant membranes
- mβ-CD:
-
Methyl β-cyclodextrin
References
Arvanitis DN, Min W, Gong Y, Heng YM, Boggs JM (2005) Two types of detergent-insoluble, glycosphingolipid/cholesterol-rich membrane domains from isolated myelin. J Neurochem 94:1696–1710
Babiychuk EB, Draeger A (2000) Annexins in cell membrane dynamics. Ca2+-regulated association of lipid microdomains. J Cell Biol 150:1113–1124
Babiychuk EB, Draeger A (2006) Biochemical characterization of detergent-resistant membranes: a systematic approach. Biochem J 397:407–416
Babiychuk EB, Monastyrskaya K, Burkhard FC, Wray S, Draeger A (2002) Modulating signaling events in smooth muscle: cleavage of annexin 2 abolishes its binding to lipid rafts. FASEB J 16:1177–1184
Bernucci L, Umana F, Llanos P, Riquelme G (2003) Large chloride channel from pre-eclamptic human placenta. Placenta 24:895–903
Berrios N, Diaz P, Riquelme G (2008) Functional incorporation of potassium channels from syncytiotrophoblast apical membrane into Xenopus laevis oocytes. Placenta 29:119
Berryman M, Gary R, Bretscher A (1995) Ezrin oligomers are major cytoskeletal components of placental microvilli: a proposal for their involvement in cortical morphogenesis. J Cell Biol 131:1231–1242
Braccia A, Villani M, Immerdal L, Niels-Christiansen LL, Nystrom BT, Hansen GH, Danielsen EM (2003) Microvillar membrane microdomains exist at physiological temperature. Role of galectin-4 as lipid raft stabilizer revealed by “superrafts”. J Biol Chem 278:15679–15684
Brown DA, Rose JK (1992) Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell 68:533–544
Chatterjee S, Smith ER, Hanada K, Stevens VL, Mayor S (2001) GPI anchoring leads to sphingolipid-dependent retention of endocytosed proteins in the recycling endosomal compartment. EMBO J 20:1583–1592
Crane JM, Tamm LK (2004) Role of cholesterol in the formation and nature of lipid rafts in planar and spherical model membranes. Biophys J 86:2965–2979
Danielsen EM, Hansen GH (2003) Lipid rafts in epithelial brush borders: atypical membrane microdomains with specialized functions. Biochim Biophys Acta 1617:1–9
Delacour D, Jacob R (2006) Apical protein transport. Cell Mol Life Sci 63:2491–2505
Fantini J, Garmy N, Mahfoud R, Yahi N (2002) Lipid rafts: structure, function and role in HIV, Alzheimers and prion diseases. Expert Rev Mol Med 2002:1–22
Gaus K, Rodriguez M, Ruberu KR, Gelissen I, Sloane TM, Kritharides L, Jessup W (2005) Domain-specific lipid distribution in macrophage plasma membranes. J Lipid Res 46:1526–1538
Hanada K, Nishijima M, Akamatsu Y, Pagano RE (1995) Both sphingolipids and cholesterol participate in the detergent insolubility of alkaline phosphatase, a glycosylphosphatidylinositol-anchored protein, in mammalian membranes. J Biol Chem 270:6254–6260
Hanono A, Garbett D, Reczek D, Chambers DN, Bretscher A (2006) EPI64 regulates microvillar subdomains and structure. J Cell Biol 175:803–813
Harder T, Gerke V (1994) The annexin II2p11(2) complex is the major protein component of the triton X–100-insoluble low-density fraction prepared from MDCK cells in the presence of Ca2+. Biochim Biophys Acta 1223:375–382
Harder T, Scheiffele P, Verkade P, Simons K (1998) Lipid domain structure of the plasma membrane revealed by patching of membrane components. J Cell Biol 141:929–942
Ikeda S, Fujimori M, Shibata S, Okajima M, Ishizaki Y, Kurihara T, Miyata Y, Iseki M, Shimizu Y, Tokumoto N, Ozaki S, Asahara T (2006) Combined immunohistochemistry of beta-catenin, cytokeratin 7, and cytokeratin 20 is useful in discriminating primary lung adenocarcinomas from metastatic colorectal cancer. BMC Cancer 6:31
Ikonen E (2001) Roles of lipid rafts in membrane transport. Curr Opin Cell Biol 13:470–477
Janich P, Corbeil D (2007) GM1 and GM3 gangliosides highlight distinct lipid microdomains within the apical domain of epithelial cells. FEBS Lett 581:1783–1787
Jimenez V, Henriquez M, Llanos P, Riquelme G (2004) Isolation and purification of human placental plasma membranes from normal and pre-eclamptic pregnancies: a comparative study. Placenta 25:422–437
Lagerholm BC, Weinreb GE, Jacobson K, Thompson NL (2005) Detecting microdomains in intact cell membranes. Annu Rev Phys Chem 56:309–336
Lichtenberg D, Goni FM, Heerklotz H (2005) Detergent-resistant membranes should not be identified with membrane rafts. Trends Biochem Sci 30:430–436
Llanos P, Henriquez M, Riquelme G (2002) A low conductance, non-selective cation channel from human placenta. Placenta 23:184–191
Macdonald JL, Pike LJ (2005) A simplified method for the preparation of detergent-free lipid rafts. J Lipid Res 46:1061–1067
Mazzone A, Tietz P, Jefferson J, Pagano R, LaRusso NF (2006) Isolation and characterization of lipid microdomains from apical and basolateral plasma membranes of rat hepatocytes. Hepatology 43:287–296
Meder D, Moreno MJ, Verkade P, Vaz WL, Simons K (2006) Phase coexistence and connectivity in the apical membrane of polarized epithelial cells. Proc Natl Acad Sci USA 103:329–334
Montalbetti N, Li Q, Wu Y, Chen XZ, Cantiello HF (2007) Polycystin–2 cation channel function in the human syncytiotrophoblast is regulated by microtubular structures. J Physiol 579:717–728
Morales FC, Takahashi Y, Kreimann EL, Georgescu MM (2004) Ezrin-radixin-moesin (ERM)-binding phosphoprotein 50 organizes ERM proteins at the apical membrane of polarized epithelia. Proc Natl Acad Sci USA 101:17705–17710
Nguyen HT, Amine AB, Lafitte D, Waheed AA, Nicoletti C, Villard C, Letisse M, Deyris V, Roziere M, Tchiakpe L, Danielle CD, Comeau L, Hiol A (2006) Proteomic characterization of lipid rafts markers from the rat intestinal brush border. Biochem Biophys Res Commun 342:236–244
Paku S, Dezso K, Kopper L, Nagy P (2005) Immunohistochemical analysis of cytokeratin 7 expression in resting and proliferating biliary structures of rat liver. Hepatology 42:863–870
Paradela A, Bravo SB, Henriquez M, Riquelme G, Gavilanes F, Gonzalez-Ros JM, Albar JP (2005) Proteomic analysis of apical microvillous membranes of syncytiotrophoblast cells reveals a high degree of similarity with lipid rafts. J Proteome Res 4:2435–2441
Pike LJ (2006) Rafts defined: a report on the Keystone symposium on lipid rafts and cell function. J Lipid Res 47:1597–1598
Rajendran L, Masilamani M, Solomon S, Tikkanen R, Stuermer CA, Plattner H, Illges H (2003) Asymmetric localization of flotillins/reggies in preassembled platforms confers inherent polarity to hematopoietic cells. Proc Natl Acad Sci USA 100:8241–8246
Rajendran L, Simons K (2005) Lipid rafts and membrane dynamics. J Cell Sci 118:1099–1102
Riquelme G, Parra M (1999) Regulation of human placental chloride channel by arachidonic acid and other cis unsaturated fatty acids. Am J Obstet Gynecol 180:469–475
Riquelme G, Stutzin A, Barros LF, Liberona JL (1995) A chloride channel from human placenta reconstituted into giant liposomes. Am J Obstet Gynecol 173:733–738
Riquelme G, Llanos P, Tischner E, Neil J, Campos B (2004) Annexin 6 modulates the maxi-chloride channel of the apical membrane of syncytiotrophoblast isolated from human placenta. J Biol Chem 279:50601–50608
Roper K, Corbeil D, Huttner WB (2000) Retention of prominin in microvilli reveals distinct cholesterol-based lipid micro-domains in the apical plasma membrane. Nat Cell Biol 2:582–592
Schroeder RJ, Ahmed SN, Zhu Y, London E, Brown DA (1998) Cholesterol and sphingolipid enhance the Triton X–100 insolubility of glycosylphosphatidylinositol-anchored proteins by promoting the formation of detergent-insoluble ordered membrane domains. J Biol Chem 273:1150–1157
Schuck S, Simons K (2004) Polarized sorting in epithelial cells: raft clustering and the biogenesis of the apical membrane. J Cell Sci 117:5955–5964
Simons K, Ehehalt R (2002) Cholesterol, lipid rafts, and disease. J Clin Invest 110:597–603
Simons K, Ikonen E (1997) Functional rafts in cell membranes. Nature 387:569–572
Simons K, Toomre D (2000) Lipid rafts and signal transduction. Nat Rev Mol Cell Biol 1:31–39
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85
Taieb N, Yahi N, Fantini J (2004) Rafts and related glycosphingolipid-enriched microdomains in the intestinal epithelium: bacterial targets linked to nutrient absorption. Adv Drug Deliv Rev 56:779–794
Tyska MJ, Mackey AT, Huang JD, Copeland NG, Jenkins NA, Mooseker MS (2005) Myosin–1a is critical for normal brush border structure and composition. Mol Biol Cell 16:2443–2457
Vallejos C, Riquelme G (2007) The maxi-chloride channel in human syncytiotrophoblast: a pathway for taurine efflux in placental volume regulation? Placenta 28:1182–1191
Vallejos C, Guerrero I, Riquelme G (2008) Potassium channels in syncytiotrophoblast: an electrophysiological challenge. Placenta 29:123
van der Goot FG, Harder T (2001) Raft membrane domains: from a liquid-ordered membrane phase to a site of pathogen attack. Semin Immunol 13:89–97
Volonte D, Galbiati F, Li S, Nishiyama K, Okamoto T, Lisanti MP (1999) Flotillins/cavatellins are differentially expressed in cells and tissues and form a hetero-oligomeric complex with caveolins in vivo. Characterization and epitope-mapping of a novel flotillin–1 monoclonal antibody probe. J Biol Chem 274:12702–12709
Wald FA, Oriolo AS, Casanova ML, Salas PJ (2005) Intermediate filaments interact with dormant ezrin in intestinal epithelial cells. Mol Biol Cell 16:4096–4107
Wiechelman KJ, Braun RD, Fitzpatrick JD (1988) Investigation of the bicinchoninic acid protein assay: identification of the groups responsible for color formation. Anal Biochem 175:231–237
Xu W, Yoon SI, Huang P, Wang Y, Chen C, Chong PL, Liu-Chen LY (2006) Localization of the kappa opioid receptor in lipid rafts. J Pharmacol Exp Ther 317:1295–1306
Acknowledgement
We are grateful to Dr. M. Pérez and the staff at the San José Hospital Maternity Unit for assistance in obtaining the biological material. We also thank Dr. V. Illanes for critical reading of the manuscript and Mr. Aldo Valdebenito for technical assistance. This research was supported by grant Fondecyt–Chile 1070695.
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Godoy, V., Riquelme, G. Distinct Lipid Rafts in Subdomains from Human Placental Apical Syncytiotrophoblast Membranes. J Membrane Biol 224, 21–31 (2008). https://doi.org/10.1007/s00232-008-9125-5
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DOI: https://doi.org/10.1007/s00232-008-9125-5