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Distinct Lipid Rafts in Subdomains from Human Placental Apical Syncytiotrophoblast Membranes

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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

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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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Authors and Affiliations

  1. Departamento de Fisiología y Biofísica, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile

    Valeria Godoy

  2. Fisiología y Biofísica, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Casilla, 70005, Santiago 7, Chile

    Gloria Riquelme

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  1. Valeria Godoy
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Correspondence to Gloria Riquelme.

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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

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