Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Oct 15;88(20):9365–9369. doi: 10.1073/pnas.88.20.9365

Hydrophobic ion transfer between membranes of adjacent hepatocytes: a possible probe of tight junction structure.

L Turin 1, P Béhé 1, I Plonsky 1, A Dunina-Barkovskaya 1
PMCID: PMC52715  PMID: 1924400

Abstract

The topology of the tight junction is probed by introducing dipicrylamine (dpa-), a lipid-soluble anion, into the membranes of hepatocyte pairs in culture. Once partitioned into the membrane, dpa- ions are free to move in the hydrophobic core of the membrane, where their mobile charges greatly increase membrane capacitance. If tight junctions are lines of membrane fusion, dpa- will cross the tight junction without traversing a polar headgroup layer. Furthermore, the electric potential across the tight junction will be equal to the difference in membrane potentials of the two cells. dpa- can therefore be expected to move electrophoretically from cell membrane to cell membrane across the junction in response to an intercellular voltage difference. Experiments performed under double whole-cell clamp show that this transfer occurs as follows: First, dpa- causes an intercellular current unrelated to gap junctions to flow in response to an intercellular voltage difference. Second, this electrophoretic removal or addition of dpa- from a cell's membrane through the tight junction must reduce or increase its dpa- content and thus its capacitance. Experiments confirm this prediction: We detect rapid, symmetric, and reversible changes in membrane capacitance in response to changes in the membrane potential of the neighboring cell. Finally, we find that hepatocyte membranes contain a negatively charged endogenous molecule that contain a negatively charged endogenous molecule that can move from cell to cell like dpa- under the influence of an intercellular potential difference. We conclude that membrane fusion occurs at tight junctions and that this hydrophobic intercellular pathway can play a role in intercellular communication.

Full text

PDF
9365

Images in this article

9366Image
on p.9366
9366Image
on p.9366

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berridge M. J. Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem. 1987;56:159–193. doi: 10.1146/annurev.bi.56.070187.001111. [DOI] [PubMed] [Google Scholar]
  2. Feltkamp C. A., Van der Waerden A. W. Junction formation between cultured normal rat hepatocytes. An ultrastructural study on the presence of cholesterol and the structure of developing tight-junction strands. J Cell Sci. 1983 Sep;63:271–286. doi: 10.1242/jcs.63.1.271. [DOI] [PubMed] [Google Scholar]
  3. Fernández J. M., Taylor R. E., Bezanilla F. Induced capacitance in the squid giant axon. Lipophilic ion displacement currents. J Gen Physiol. 1983 Sep;82(3):331–346. doi: 10.1085/jgp.82.3.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Frankel J. Positional information in unicellular organisms. J Theor Biol. 1974 Oct;47(2):439–481. doi: 10.1016/0022-5193(74)90209-4. [DOI] [PubMed] [Google Scholar]
  5. Hertzberg E. L., Spray D. C., Bennett M. V. Reduction of gap junctional conductance by microinjection of antibodies against the 27-kDa liver gap junction polypeptide. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2412–2416. doi: 10.1073/pnas.82.8.2412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hirsch M., Montcourrier P. A further iconographic argument in favour of the 'offset two-fibril' model of the tight junction. Cell Biol Int Rep. 1983 Jun;7(6):406–406. doi: 10.1016/0309-1651(83)90128-5. [DOI] [PubMed] [Google Scholar]
  7. Johnston M. F., Simon S. A., Ramón F. Interaction of anaesthetics with electrical synapses. Nature. 1980 Jul 31;286(5772):498–500. doi: 10.1038/286498a0. [DOI] [PubMed] [Google Scholar]
  8. Kachar B., Reese T. S. Evidence for the lipidic nature of tight junction strands. Nature. 1982 Apr 1;296(5856):464–466. doi: 10.1038/296464a0. [DOI] [PubMed] [Google Scholar]
  9. Kikkawa U., Nishizuka Y. The role of protein kinase C in transmembrane signalling. Annu Rev Cell Biol. 1986;2:149–178. doi: 10.1146/annurev.cb.02.110186.001053. [DOI] [PubMed] [Google Scholar]
  10. Meyer H. W. Tight junction strands are lipidic cylinders. Naturwissenschaften. 1983 May;70(5):251–252. doi: 10.1007/BF00405446. [DOI] [PubMed] [Google Scholar]
  11. Mittelman L. A., Sharovskaya J. J., Vasiliev J. M. Toxic effect of 7,12-dimethylbenz-alpha-anthracene on neoplastic cells grown in mixed cultures with normal fibroblasts. Int J Cancer. 1972 Nov;10(3):667–674. doi: 10.1002/ijc.2910100326. [DOI] [PubMed] [Google Scholar]
  12. Nichols G. E., Borgman C. A., Young W. W., Jr On tight junction structure: Forssman glycolipid does not flow between MDCK cells in an intact epithelial monolayer. Biochem Biophys Res Commun. 1986 Aug 14;138(3):1163–1169. doi: 10.1016/s0006-291x(86)80404-1. [DOI] [PubMed] [Google Scholar]
  13. Pickar A. D., Brown W. C. Capacitance of bilayers in the presence of lipophilic ions. Biochim Biophys Acta. 1983 Aug 24;733(1):181–185. doi: 10.1016/0005-2736(83)90104-9. [DOI] [PubMed] [Google Scholar]
  14. Pinto da Silva P., Kachar B. On tight-junction structure. Cell. 1982 Mar;28(3):441–450. doi: 10.1016/0092-8674(82)90198-2. [DOI] [PubMed] [Google Scholar]
  15. Turin L., Warner A. E. Intracellular pH in early Xenopus embryos: its effect on current flow between blastomeres. J Physiol. 1980 Mar;300:489–504. doi: 10.1113/jphysiol.1980.sp013174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Warner A. E., Guthrie S. C., Gilula N. B. Antibodies to gap-junctional protein selectively disrupt junctional communication in the early amphibian embryo. Nature. 1984 Sep 13;311(5982):127–131. doi: 10.1038/311127a0. [DOI] [PubMed] [Google Scholar]
  17. Wolpert L. Positional information revisited. Development. 1989;107 (Suppl):3–12. doi: 10.1242/dev.107.Supplement.3. [DOI] [PubMed] [Google Scholar]
  18. van Meer G., Gumbiner B., Simons K. The tight junction does not allow lipid molecules to diffuse from one epithelial cell to the next. Nature. 1986 Aug 14;322(6080):639–641. doi: 10.1038/322639a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES