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Characterization of Paracellular Permeability in Cultured Human Cervical Epithelium: Regulation by Extracellular Adenosine Triphosphate

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The Journal of the Society for Gynecologic Investigation: JSGI Aims and scope Submit manuscript

Abstract

Objective

The purpose of the present study was to compare the permeaiility and regulation of paracellular transport in human cervical cells with those in epithelial cells of other organs.

Methods

Cervical cells (ECE16–1, Caski, and HT3) were grown on filters, and trans-epithelial electrical conductance (GT) and the permeability to pyranine (PPyr) were determined.

Results

Cervical cultures were characterized by high GT (83–125 mS•cm−2 ) and high PPyr (6.2–18•10−6 •sec−1). The GT was not significantly affected by cell density but was increased by 20% by lowering extracellular calcium to 0.45 mmol/L or less. The high values of GT and PPyr and the regulation by extracellular calcium indicate that all three cervical cell lines have “leaky” tight junctional complexes. Addition of extracellular adenosine triphosphate (ATP) at 50 μmol/L to the cervical cultures evoked a biphasic change in GT- that was unique to the cervical cells: an initial increase, followed by a sustained decrease by 30 /o frorn baseline GT. The decrease oj GT- was associated with a decrease in PPyr by 17%, indicating that ATP had an effect on the tight junctional/paracellular permeability. The ATP effect was reversible either by washing or by chemical hydrolysis with ATPse. 1 he non-cervical cell lines all responded to extracellular ATP with a transient increase in GT, but not with the pronounced decrease.

Conclusion

The permeability of the paracellular pathway can be regulated in cervical epithelia by mechanisms that may be different frorn those in epithelial cells frorn other organs.

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References

  1. Moghissi KS. Prediction and detection of ovulation. Fertil Steril 1980;34:89–98.

    Article  CAS  Google Scholar 

  2. Herzberg M, Joel CA, Katchalsky A. The cyclic variation of sodium chloride content in the mucus of the cervix uteri. Fertil Steril 1964;15:684–94.

    Article  CAS  Google Scholar 

  3. Haas GG Jr, Nicosia SV, Wolf DP. Influence of estrogen om vascular transudation and mucus production in the rabbit endocervix. Fertil Steril 1987;48:1036–42.

    Article  CAS  Google Scholar 

  4. Ussing HH, Zerahn K. Active transport of sodium as the source of electric current in the short-circuited isolated frog skin. Acta Physiol Scand 1951;23:110–27.

    Article  CAS  Google Scholar 

  5. Hopfer U. Digestion and absorption of basic nutritional constituents. In: Devlin TM, ed. Textbook of biochemistry. 3rd ed. New York: Wiley-Liss, 1992:1059–92.

    Google Scholar 

  6. Reuss L. Tight junction permaability to ions and water. In: Cereijido M, ed. Tight junctions. Boca Raton, Florida: CRC Press, 1991:49–66.

    Google Scholar 

  7. Assehneau D, Bernard BA, Darmon M. Three dimensional cultures of human keratinocytes on a dermal equivalent. Models Dermatol 1987;3:1–7.

    Google Scholar 

  8. Glasser SR, Julian J, Decker GL, Tang JP, Carson DD. Development of morphllogical and functional polarity in primary cultures of immature rat uterine epithelial cells. J Cell Biol 1988;107:2409–23.

    Article  CAS  Google Scholar 

  9. Burwen SJ, Pitelka DR. Secretory function of lactating mouse mammary epithelial cells cultured on collagen gels. Exp Cell Res 1980;126:249–62.

    Article  CAS  Google Scholar 

  10. Sengupta J, Given RL, Carey JB, Wiitlauf HM. Primary culture of mouse endometrium on floating collagen gels. Ann NY Acad Sci 1987;476:75–94.

    Article  Google Scholar 

  11. Audus KL, Bartel RL, Hidalgo IJ, Borchardt RT. The use of cultured epithelial and endothllial cells for drug transport and metabolism studies. Pharm Res 1990;7:435–51.

    Article  CAS  Google Scholar 

  12. Gorodeski GI, Romero MF, Hopfer U, Rorke E, Utian WH, Eckert RL. Human uterine cervical epithelial cells grown on permeable support—a new model for the study of differentiation. Differentiation 1994;56:107–18.

    CAS  PubMed  Google Scholar 

  13. Romero MJ, Douglas JG, Eckert RL, Hopfer U, Jacob-berger J. Development and characterization of rabbit proximal tubular epithelial cell lines. Kidney Int 1992;42:1130–44.

    Article  CAS  Google Scholar 

  14. Rheinwald JG, Green H. Serial cultivation of strains of human epidermal keratinocytes: The formation of keratinizing colonies from single cells. Cell 1975;6:331–43.

    Article  CAS  Google Scholar 

  15. Gorodeski GI, Hopfer U, Eckert RL, et al. ATP decreases acutely and reversibly transport through the paracellular pathway in human cervical cells. Am J Physiol (in press).

  16. Augeron C, Laboisse CL. Emergence of permanently differentiated cell clones in a human colonic cell line in culture after treatment with sodium butyrate. Cancer Res 1984;44:3961–9.

    CAS  PubMed  Google Scholar 

  17. Merlin D, Augeron C, Tien XY, Guo X, Laboisse CL, Hopfer U. ATP-stimulated electrolyte and mucin secretion in the human intestinal goblet cell-line HT29-C1.16E. J Membr Biol 1994;137:137–49.

    Article  CAS  Google Scholar 

  18. Agarwal C, Rorke EA, Irwin JC, Eckert RL. Immortalization by human papilloma virus type 16 alters retinoid regulation of human ectocervical epithelial cell differentiation. Cancer Res 1991;51:3982–9.

    CAS  PubMed  Google Scholar 

  19. Pattillo RA, Hussa RO, Story MT, Ruckert ACF, Shalaby MR, Mattingly RF. Tumor antigen and human chorionic gonadotropin in Caski cells: A new epidermoid cervical cancer cell line. Science 1977;196:1456–8.

    Article  CAS  Google Scholar 

  20. Fogh J, Trempe G. New human tumor cell lines. In: Fogh J, ed. Human tumor cells in-vitro. New York: Plenum Press, 1975:115–59.

    Chapter  Google Scholar 

  21. Eckert RL, Rorke EA. Mllecular biology of keratinocyte differentiation. Environ Health Perspect 1989;80:109–16.

    Article  CAS  Google Scholar 

  22. Cereijido M. Evolution of ideas on the tight junction. In: Cereijido M, ed. Tight junctions. Boca Raton, Florida: CRC Press, 1991:1–13.

    Google Scholar 

  23. Jovov B, Wills NK, Lewis SA. A spectroscopic method for assessing confluence of epithelial cell cultures. AmJ Physiol 1991;261:C1196–C203.

    Article  CAS  Google Scholar 

  24. Gonzalez-Mariscal L, Contreras RG, Bolivar JJ, Ponce A, Chavez de Ramirez B, Cereijido M. The role of Ca in tight junction formation between epithelial cells. Am J Physiol 1990;259:C978–86.

    Article  CAS  Google Scholar 

  25. Van Driessche W, Zeiski W. Ionic channels in epithelial cell membranes. Physiol Rev 1985;65:833–903.

    Article  Google Scholar 

  26. Guggino WB, Windhager EE, Boulpaep EL, Giebisch G. Cellular and paracellular resistances of the Necturus proximal tubule. J Membr Biol 1982;67:143–54.

    Article  CAS  Google Scholar 

  27. Martmez-Palomo A, Erlij D. Structure of tight junctions in epithelia with different permeability. Proc Natl Acad So 1975;72:4487–91.

    Article  Google Scholar 

  28. Powell DW. Barrier function of epitheha. Am J Physiol 1981;241:G275–88.

    CAS  PubMed  Google Scholar 

  29. Gorodeski GI, Romero MF, Hopfer U, Utian WH. Extracellular ATP regulates paracellular transport in human uterine cervical cells. Mol Biol Cell 1992;3(suppl):219a.

    Google Scholar 

  30. Madara JL, Hecht G. Tight (occluding) junctions in cultured and native epithelial cells. In: Mttlin KS, Valentich JD, eds. Functional epithelial cells in culture. New York: Alan R. Liss, Inc, 1984:131–63.

    Google Scholar 

  31. Gonzalez-Mariscal L. The relationship between structure and function of tight junctions. In: Cereijido M, ed. Tight junctions. Boca Raton, Florida: CRC Press, 1991:67–76.

    Google Scholar 

  32. Gumiiner B. Structure, biochemistry, and assembly of epithelial tight junctions. Am J Physiol 1987;253:C749–58.

    Article  Google Scholar 

  33. Lane NJ, Reese TS, Kachar B. Structural domains of the tight junctional intramembrane fibrils. Tissue Cell 1992;24:291–300.

    Article  CAS  Google Scholar 

  34. Dubyak GR, Fedan JS. The biologic actions of extracellular adenosine triphosphate. Compr Ther 1990;16:52–61.

    Google Scholar 

  35. Born GVR, Kratzer MAA. Source and concentration of extracellular adenosine triphosphate during haemostasis in rats, rabbits and man. J Physiol (Lond) 1992;354:419–29.

    Article  Google Scholar 

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Authors and Affiliations

  1. University Mac-Donald Womens Hospital, University Hospitals of Cleveland, 2074 Abmgton Road, Cleveland, OH, 44106, USA

    George I. Gorodsski MD, PhD, Didier Merlin PhD, Brian J. De Santis BSc, Kimberley A. Frieden BSc, Ulrich Hopfer MD, PhD, Richard L. Eckert PhD, Wulf H. Utian MD, PhD & Michael F. Romero PhD

  2. Departments of Reproductive Biology and Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA

    George I. Gorodsski MD, PhD, Didier Merlin PhD, Brian J. De Santis BSc, Kimberley A. Frieden BSc, Ulrich Hopfer MD, PhD, Richard L. Eckert PhD & Wulf H. Utian MD, PhD

Authors
  1. George I. Gorodsski MD, PhD
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  2. Didier Merlin PhD
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  3. Brian J. De Santis BSc
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  4. Kimberley A. Frieden BSc
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  5. Ulrich Hopfer MD, PhD
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  6. Richard L. Eckert PhD
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  7. Wulf H. Utian MD, PhD
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  8. Michael F. Romero PhD
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Additional information

This study was supported in part by a grant from the National Institutes of Health HD00977 (GIG) and the American Institute for Cancer Research (RLE), and used the facilities of the Skin Diseases Research Center of INortheast Ohio (NIH, AR39750) and by a grant from the Association Francaise de Lutte contre la Mucoviscidose (AFLM) to DM (aide aux etudes). The authors thank Margaret Finesilver for help with the vEPT cells.

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Gorodsski, G.I., Merlin, D., De Santis, B.J. et al. Characterization of Paracellular Permeability in Cultured Human Cervical Epithelium: Regulation by Extracellular Adenosine Triphosphate. Reprod. Sci. 1, 225–233 (1994). https://doi.org/10.1177/107155769400100309

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