Proteins of the apical and basal plasma membranes of the human placental syncytiotrophoblast: Immunochemical and electrophoretic studies

Vanderpuye, Oluseyi A.; Smith, Carl H.

doi:10.1016/0143-4004(87)90030-0

Cited by 14 publications

(2 citation statements)

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“…Such a structural surface polarity is a common property of epithelia, which function in vectorial transport (Simons and Fuller, 1985) . These data are consistent with Vanderpuye et al (1987Vanderpuye et al ( , 1988, who reported structural differences between apical and basal plasma membranes isolated from After incubation, cells were incubated with DAB, with or without H202 as indicated. After DAB cytochemistry, cells were processed for SDS-PAGE as described in Materials and Methods.…”

Section: Discussionsupporting

confidence: 90%

Apical and basolateral transferrin receptors in polarized BeWo cells recycle through separate endosomes.

Cerneus

Ende

1991

The Journal of Cell Biology

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Abstract. Contrary to most other epithelia, trophoblasts in the human placenta, which form the physical barrier between the fetal and the maternal blood circulation, express high numbers of transferrin receptors on their apical cell surface. This study describes the establishment of a polarized trophoblast-like cell line BeWo, which exhibit a high expression of transferrin receptors on the apex of the cells. Cultured on permeable filter supports, BeWo cells formed a polarized monolayer with microvilli on their apical cell surface. Across the monolayer a transepithelial resistance developed of -600 SZ-cm2 within 4 d. Depletion of Call from the medium decreased the resistance to background levels, showing its dependence on the integrity of tight junctions . Within the same period of time the secretion of proteins became polarized . In addition, the compositions of integral membrane proteins at the apical and basolateral plasma membrane domains were distinct as determined by domain-selective iodination . Similar to placental trophoblasts, binding of 125 1-labeled transferrin to BeWo monolayers revealed T 11E plasma membrane of epithelial cells has differentiated into an apical and a basolateral domain, which are morphologically and biochemicallY distinct . The establishment and maintenance of this cell surface polarity is achieved by several mechanisms. These include proper cell-substrate attachment, intensive cell-cell contact, where tight junctions prevent lateral diffusion ofproteins and lipids from one domain to the other, and targeting of newly synthesized proteins to the appropriate domain (Rodriguez-Boulan and Nelson, 1989;Simons and Wandinger-Ness, 1990) . Transcytosis from both domains may contribute to the formation of plasma membrane domains (Hubbard, 1989), but may also compromise the maintenance of cell surface polarity. However, efficient sorting of recycling proteins from proteins which are transcytosed (Mostov and Simister, 1985 ;Hubbard, 1989;Rodman et al., 1990), as well as separate apical and basolateral early endosomes (Oliver, 1982;Von Bonsdorff et al., 1985;Bomsel et al., 1989;Parton et al ., 1989), prevent intracellular mixing of proteins from different domains .© The Rockefeller University Press, 0021-9525/91/09/1149/10 $2.00 The Journal of Cell Biology, Volume 114, Number 6, September 19911149-1158 1149 that the transferrin receptor was expressed at both plasma membrane domains. Apical and basolateral transferrin receptors were found in a 1:2 surface ratio and exhibited identical dissociation constants and molecular weights . After uptake, transferrin recycled predominantly to the domain of administration, indicating separate recycling pathways from the apical and basolateral domain . This was confirmed by using diaminobenzidine cytochemistry, a technique by which colocalization of endocytosed 125 1-labeled and HRR conjugated transferrin can be monitored . No mixing of the two types of ligands was observed, when both ligands were simultaneously internalized for 10 or 60 min from opposite do...

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Section: Discussionsupporting

confidence: 90%

Apical and basolateral transferrin receptors in polarized BeWo cells recycle through separate endosomes.

Cerneus

Ende

1991

The Journal of Cell Biology

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“…The infection is relatively inefficient even in the presence of a high virus inoculum and progresses slowly, and progeny virus remains predominantly cell associated with limited release into the supernatant (22). However, ST cells are epithelial cells with distinct apical and basal surfaces (7,12,23,26,42,51). Since release of virus in a basal direction could result in direct transmission to fetal tissue, a more physiologic assessment of a productive HCMV infection of ST would be measurement of basal release.…”

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confidence: 99%

Polarized Release of Human Cytomegalovirus from Placental Trophoblasts

Hemmings

Guilbert

2002

J Virol

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Human cytomegalovirus (HCMV) is a ubiquitous infectious pathogen that, when transmitted to the fetus in utero, can result in numerous sequelae, including late-onset sensorineural damage. The villous trophoblast, the cellular barrier between maternal blood and fetal tissue in the human placenta, is infected by HCMV in vivo. Primary trophoblasts cultured on impermeable surfaces can be infected by HCMV, but release of progeny virus is delayed and minimal. It is not known whether these epithelial cells when fully polarized can release HCMV and, if so, if release is from the basal membrane surface toward the fetus. We therefore ask whether, and in which direction, progeny virus release occurs from HCMV-infected trophoblasts cultured on semipermeable (3.0-m-pore-size) membranes that allow functional polarization. We show that infectious HCMV readily diffuses across cell-free 3.0-m-pore-size membranes and that apical infection of confluent and multilayered trophoblasts cultured on these membranes reaches cells at the membrane surface. Using two different infection and culture protocols, we found that up to 20% of progeny virus is released but that <1% of released virus is detected in the basal culture chamber. These results suggest that very little, if any, HCMV is released from an infected villous trophoblast into the villous stroma where the virus could ultimately infect the fetus.Human cytomegalovirus (HCMV) is endemic, infecting 40 to 80% of urban populations in developed countries (10). The incidence of congenital HCMV infection is between 0.2 and 2.2% of all live-born infants. Such infections are one of the most common causes of mental retardation and nonhereditary sensorineural deafness in children (46,48). Only 40% of pregnant women with primary HCMV infections give birth to infected infants (47) suggesting an effective fetal barrier, either physical or immunological. How HCMV is transmitted to the fetus during pregnancy is unknown; however, congenital infections are associated with chronic villitis (38, 41) and infection of the placenta (1, 4, 16, 32, 36-39, 41, 44, 45), which may also act as a viral reservoir (19). Passage through the placenta (2, 3) could occur in two directions: through the cytotrophoblast (CT) columns in anchoring villi or across the villous syncytiotrophoblast (ST) in the intervillous space.Infection originating in the uterine wall could lead to infection of extravillous CT involved in either interstitial or endovascular invasion. Infection could then progress in a cell-to-cell manner through the CT columns of anchoring villi to the chorionic stroma and eventually infect the fetus (15). Although this route is possible during a reactivated uterine infection, endovascular CT comes into direct contact with maternal blood and therefore could also be exposed to a primary infection.The ST lines all chorionic villi within the intervillous space, thereby interfacing maternal blood and fetal tissues. The ST is thus positioned to either physically prevent or to actively participate in the dissemina...

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Basic Structure of the Villous Trees

Castellucci¹,

Kaufmann²

Pathology of the Human Placenta

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Proteins of the apical and basal plasma membranes of the human placental syncytiotrophoblast: Immunochemical and electrophoretic studies

Cited by 14 publications

References 40 publications

Apical and basolateral transferrin receptors in polarized BeWo cells recycle through separate endosomes.

Apical and basolateral transferrin receptors in polarized BeWo cells recycle through separate endosomes.

Polarized Release of Human Cytomegalovirus from Placental Trophoblasts

Basic Structure of the Villous Trees

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