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Properties of two multisubstate Cl− channels from human syncytiotrophoblast reconstituted on planar lipid bilayers

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Abstract

We describe the first successful reconstitution of placental ionic channels on planar lipid bilayers. An apical plasma membrane-enriched vesicle fraction from human syncytiotrophoblast at term was prepared by following isotonic agitation, differential centrifugation, and Mg2+-induced selective precipitation of nonapical membranes, and its purity was assessed by biochemical and morphological marker analysis. We have already reported that, unlike previous patch-clamp studies, nonselective cation channels were incorporated in most cases, a result consistent with the higher permeability for cations as compared with Cl− and with the low apical membrane potential difference at term revealed by fluorescent probe partition studies, and microelectrode techniques. In this paper, we report that Cl−-selective channels were incorporated in 4% of successful reconstitutions (14 out of 353) and that their analysis revealed two types of activity. One of them was consistent with a voltagedependent, 100-pS channel while the other was consistent with the lateral association of 47-pS conductive units, giving rise to multibarrelled, DIDS-sensitive channels of variable conductance (300 to 650 pS). The latter displayed a very complex behavior which included cooperative gating of conductive units, long-lived substates, voltage-dependent entry into an apparent inactivated state, and flickering activity. The role of the reported Cl− channels in transplacental ion transport and/or syncytium homeostasis remains to be determined.

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Authors and Affiliations

  1. Departamento de Química Analítica y Fisicoquímica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, 1113, Buenos Aires, Argentina

    C. Grosman & I. L. Reisin

  2. Servicio de Microscopía Electrónica, Inst. Nac. de Microbiol, Dr. C.G. Malbrán, Av. Vélez Sarsfield 563, 1281, Buenos Aires, Argentina

    M. I. Mariano & J. P. Bozzini

Authors
  1. C. Grosman
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  2. M. I. Mariano
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  3. J. P. Bozzini
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  4. I. L. Reisin
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Grosman, C., Mariano, M.I., Bozzini, J.P. et al. Properties of two multisubstate Cl− channels from human syncytiotrophoblast reconstituted on planar lipid bilayers. J. Membrane Biol. 157, 83–95 (1997). https://doi.org/10.1007/s002329900218

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  • DOI: https://doi.org/10.1007/s002329900218

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