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Epithelial Barrier Modulation by a Channel Forming Peptide

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Abstract

NC-1059 is a synthetic channel-forming peptide that provides for ion transport across, and transiently reduces the barrier integrity of, cultured epithelial monolayers derived from canine kidney (MDCK cells). Experiments were conducted to determine whether epithelial cells derived from other sources were similarly affected. Epithelial cells derived from human intestine (T-84), airway (Calu-3), porcine intestine (IPEC-J2) and reproductive duct (PVD9902) were grown on permeable supports. Basal short circuit current (I sc) was <3 μA cm−2 for T-84, IPEC-J2 and PVD9902 cell monolayers and <8 μA cm−2 for Calu-3 cells. Apical NC-1059 exposure caused, in all cell types, an increase in I sc to >15 μA cm−2, indicative of net anion secretion or cation absorption, which was followed by an increase in transepithelial conductance (in mS cm−2: T-84, 1.6 to 62; PVD9902, 0.2 to 51; IPEC-J2, 0.3 to 26; Calu-3, 2.3 to 13). These results are consistent with the peptide affecting transcellular ion movement, with a likely effect also on the paracellular route. NC-1059 exposure increased dextran permeation when compared to basal permeation, which documents an effect on the paracellular pathway. In order to evaluate membrane ion channels, experiments were conducted to study the dose dependence and stability of the NC-1059-induced membrane conductance in Xenopus laevis oocytes. NC-1059 induced a dose-dependent increase in oocyte membrane conductance that remained stable for greater than 2 h. The results demonstrate that NC-1059 increases transcellular conductance and paracellular permeation in a wide range of epithelia. These effects might be exploited to promote drug delivery across barrier epithelia.

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Acknowledgements

We thank Gary Radke and Ryan Carlin for their technical assistance. This article is contribution 08-76-J from the Kansas Agriculture Experiment Station. This study was supported in part by Public Health Service grants GM 066620 and GM 074096 (both to J. M. T.).

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Authors and Affiliations

  1. Yale School of Medicine Cellular and Molecular Physiology, New Haven, CT, USA

    Suma Somasekharan

  2. Department of Biochemistry, Kansas State University, Manhattan, KS, USA

    Robert Brandt, Takeo Iwamoto & John M. Tomich

  3. Department of Anatomy and Physiology, Kansas State University, 1600 Denison Avenue, Coles Hall 228, Manhattan, KS, 66506, USA

    Bruce D. Schultz

Authors
  1. Suma Somasekharan
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  2. Robert Brandt
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  3. Takeo Iwamoto
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  4. John M. Tomich
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  5. Bruce D. Schultz
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Correspondence to Bruce D. Schultz.

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Somasekharan, S., Brandt, R., Iwamoto, T. et al. Epithelial Barrier Modulation by a Channel Forming Peptide. J Membrane Biol 222, 17–30 (2008). https://doi.org/10.1007/s00232-008-9099-3

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  • DOI: https://doi.org/10.1007/s00232-008-9099-3

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