Skip to main content
NCBI home page
Molecular Biology of the Cell logoLink to Molecular Biology of the Cell
. 1993 Jan;4(1):7–20. doi: 10.1091/mbc.4.1.7

Connexin40, a component of gap junctions in vascular endothelium, is restricted in its ability to interact with other connexins.

R Bruzzone 1, J A Haefliger 1, R L Gimlich 1, D L Paul 1
PMCID: PMC300896  PMID: 8382974

Abstract

The cellular distribution of connexin40 (Cx40), a newly cloned gap junction structural protein, was examined by immunofluorescence microscopy using two different specific anti-peptide antibodies. Cx40 was detected in the endothelium of muscular as well as elastic arteries in a punctate pattern consistent with the known distribution of gap junctions. However, it was not detected in other cells of the vascular wall. By contrast, Cx43, another connexin present in the cardiovascular system, was not detected in endothelial cells of muscular arteries but was abundant in the myocardium and aortic smooth muscle. We have tested the ability of these connexins to interact functionally. Cx40 was functionally expressed in pairs of Xenopus oocytes and induced the formation of intercellular channels with unique voltage dependence. Unexpectedly, communication did not occur when oocytes expressing Cx40 were paired with those expressing Cx43, although each could interact with a different connexin, Cx37, to form gap junction channels in paired oocytes. These findings indicate that establishment of intercellular communication can be spatially regulated by the selective expression of different connexins and suggest a mechanism that may operate to control the extent of communication between cells.

Full text

PDF
7

Images in this article

9Image
on p.9
11Image
on p.11
12Image
on p.12
14Image
on p.14

Selected References

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

  1. Baker C., Holland D., Edge M., Colman A. Effects of oligo sequence and chemistry on the efficiency of oligodeoxyribonucleotide-mediated mRNA cleavage. Nucleic Acids Res. 1990 Jun 25;18(12):3537–3543. doi: 10.1093/nar/18.12.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrio L. C., Suchyna T., Bargiello T., Xu L. X., Roginski R. S., Bennett M. V., Nicholson B. J. Gap junctions formed by connexins 26 and 32 alone and in combination are differently affected by applied voltage. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8410–8414. doi: 10.1073/pnas.88.19.8410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett M. V., Barrio L. C., Bargiello T. A., Spray D. C., Hertzberg E., Sáez J. C. Gap junctions: new tools, new answers, new questions. Neuron. 1991 Mar;6(3):305–320. doi: 10.1016/0896-6273(91)90241-q. [DOI] [PubMed] [Google Scholar]
  4. Beyer E. C., Kistler J., Paul D. L., Goodenough D. A. Antisera directed against connexin43 peptides react with a 43-kD protein localized to gap junctions in myocardium and other tissues. J Cell Biol. 1989 Feb;108(2):595–605. doi: 10.1083/jcb.108.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beyer E. C., Paul D. L., Goodenough D. A. Connexin43: a protein from rat heart homologous to a gap junction protein from liver. J Cell Biol. 1987 Dec;105(6 Pt 1):2621–2629. doi: 10.1083/jcb.105.6.2621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beyer E. C., Reed K. E., Westphale E. M., Kanter H. L., Larson D. M. Molecular cloning and expression of rat connexin40, a gap junction protein expressed in vascular smooth muscle. J Membr Biol. 1992 Apr;127(1):69–76. doi: 10.1007/BF00232759. [DOI] [PubMed] [Google Scholar]
  7. Burt J. M., Spray D. C. Single-channel events and gating behavior of the cardiac gap junction channel. Proc Natl Acad Sci U S A. 1988 May;85(10):3431–3434. doi: 10.1073/pnas.85.10.3431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bény J. L., Connat J. L. An electron-microscopic study of smooth muscle cell dye coupling in the pig coronary arteries. Role of gap junctions. Circ Res. 1992 Jan;70(1):49–55. doi: 10.1161/01.res.70.1.49. [DOI] [PubMed] [Google Scholar]
  9. Bény J. L., Gribi F. Dye and electrical coupling of endothelial cells in situ. Tissue Cell. 1989;21(6):797–802. doi: 10.1016/0040-8166(89)90030-x. [DOI] [PubMed] [Google Scholar]
  10. Chanson M., Orci L., Meda P. Extent and modulation of junctional communication between pancreatic acinar cells in vivo. Am J Physiol. 1991 Jul;261(1 Pt 1):G28–G36. doi: 10.1152/ajpgi.1991.261.1.G28. [DOI] [PubMed] [Google Scholar]
  11. Dahl G., Miller T., Paul D., Voellmy R., Werner R. Expression of functional cell-cell channels from cloned rat liver gap junction complementary DNA. Science. 1987 Jun 5;236(4806):1290–1293. doi: 10.1126/science.3035715. [DOI] [PubMed] [Google Scholar]
  12. Dermietzel R., Traub O., Hwang T. K., Beyer E., Bennett M. V., Spray D. C., Willecke K. Differential expression of three gap junction proteins in developing and mature brain tissues. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10148–10152. doi: 10.1073/pnas.86.24.10148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ebihara L., Beyer E. C., Swenson K. I., Paul D. L., Goodenough D. A. Cloning and expression of a Xenopus embryonic gap junction protein. Science. 1989 Mar 3;243(4895):1194–1195. doi: 10.1126/science.2466337. [DOI] [PubMed] [Google Scholar]
  14. Eghbali B., Kessler J. A., Spray D. C. Expression of gap junction channels in communication-incompetent cells after stable transfection with cDNA encoding connexin 32. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1328–1331. doi: 10.1073/pnas.87.4.1328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Epstein M. L., Gilula N. B. A study of communication specificity between cells in culture. J Cell Biol. 1977 Dec;75(3):769–787. doi: 10.1083/jcb.75.3.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Findlay I., Petersen O. H. The extent of dye-coupling between exocrine acinar cells of the mouse pancreas. The dye-coupled acinar unit. Cell Tissue Res. 1983;232(1):121–127. doi: 10.1007/BF00222378. [DOI] [PubMed] [Google Scholar]
  17. Fishman G. I., Spray D. C., Leinwand L. A. Molecular characterization and functional expression of the human cardiac gap junction channel. J Cell Biol. 1990 Aug;111(2):589–598. doi: 10.1083/jcb.111.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Giaume C., Kado R. T., Korn H. Voltage-clamp analysis of a crayfish rectifying synapse. J Physiol. 1987 May;386:91–112. doi: 10.1113/jphysiol.1987.sp016524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gimlich R. L., Kumar N. M., Gilula N. B. Differential regulation of the levels of three gap junction mRNAs in Xenopus embryos. J Cell Biol. 1990 Mar;110(3):597–605. doi: 10.1083/jcb.110.3.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gimlich R. L., Kumar N. M., Gilula N. B. Sequence and developmental expression of mRNA coding for a gap junction protein in Xenopus. J Cell Biol. 1988 Sep;107(3):1065–1073. doi: 10.1083/jcb.107.3.1065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Goodenough D. A., Paul D. L., Jesaitis L. Topological distribution of two connexin32 antigenic sites in intact and split rodent hepatocyte gap junctions. J Cell Biol. 1988 Nov;107(5):1817–1824. doi: 10.1083/jcb.107.5.1817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Haefliger J. A., Bruzzone R., Jenkins N. A., Gilbert D. J., Copeland N. G., Paul D. L. Four novel members of the connexin family of gap junction proteins. Molecular cloning, expression, and chromosome mapping. J Biol Chem. 1992 Jan 25;267(3):2057–2064. [PubMed] [Google Scholar]
  23. Hennemann H., Suchyna T., Lichtenberg-Fraté H., Jungbluth S., Dahl E., Schwarz J., Nicholson B. J., Willecke K. Molecular cloning and functional expression of mouse connexin40, a second gap junction gene preferentially expressed in lung. J Cell Biol. 1992 Jun;117(6):1299–1310. doi: 10.1083/jcb.117.6.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hoh J. H., John S. A., Revel J. P. Molecular cloning and characterization of a new member of the gap junction gene family, connexin-31. J Biol Chem. 1991 Apr 5;266(10):6524–6531. [PubMed] [Google Scholar]
  25. Kadle R., Zhang J. T., Nicholson B. J. Tissue-specific distribution of differentially phosphorylated forms of Cx43. Mol Cell Biol. 1991 Jan;11(1):363–369. doi: 10.1128/mcb.11.1.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kalimi G. H., Lo C. W. Communication compartments in the gastrulating mouse embryo. J Cell Biol. 1988 Jul;107(1):241–255. doi: 10.1083/jcb.107.1.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kam E., Hodgins M. B. Communication compartments in hair follicles and their implication in differentiative control. Development. 1992 Feb;114(2):389–393. doi: 10.1242/dev.114.2.389. [DOI] [PubMed] [Google Scholar]
  28. Kanter H. L., Saffitz J. E., Beyer E. C. Cardiac myocytes express multiple gap junction proteins. Circ Res. 1992 Feb;70(2):438–444. doi: 10.1161/01.res.70.2.438. [DOI] [PubMed] [Google Scholar]
  29. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kumar N. M., Gilula N. B. Cloning and characterization of human and rat liver cDNAs coding for a gap junction protein. J Cell Biol. 1986 Sep;103(3):767–776. doi: 10.1083/jcb.103.3.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lal R., Arnsdorf M. F. Voltage-dependent gating and single-channel conductance of adult mammalian atrial gap junctions. Circ Res. 1992 Sep;71(3):737–743. doi: 10.1161/01.res.71.3.737. [DOI] [PubMed] [Google Scholar]
  32. Larson D. M., Haudenschild C. C., Beyer E. C. Gap junction messenger RNA expression by vascular wall cells. Circ Res. 1990 Apr;66(4):1074–1080. doi: 10.1161/01.res.66.4.1074. [DOI] [PubMed] [Google Scholar]
  33. Lo C. W., Gilula N. B. Gap junctional communication in the post-implantation mouse embryo. Cell. 1979 Oct;18(2):411–422. doi: 10.1016/0092-8674(79)90060-6. [DOI] [PubMed] [Google Scholar]
  34. Meda P., Chanson M., Pepper M., Giordano E., Bosco D., Traub O., Willecke K., el Aoumari A., Gros D., Beyer E. C. In vivo modulation of connexin 43 gene expression and junctional coupling of pancreatic B-cells. Exp Cell Res. 1991 Feb;192(2):469–480. doi: 10.1016/0014-4827(91)90066-4. [DOI] [PubMed] [Google Scholar]
  35. Methfessel C., Witzemann V., Takahashi T., Mishina M., Numa S., Sakmann B. Patch clamp measurements on Xenopus laevis oocytes: currents through endogenous channels and implanted acetylcholine receptor and sodium channels. Pflugers Arch. 1986 Dec;407(6):577–588. doi: 10.1007/BF00582635. [DOI] [PubMed] [Google Scholar]
  36. Michalke W., Loewenstein W. R. Communication between cells of different type. Nature. 1971 Jul 9;232(5306):121–122. doi: 10.1038/232121b0. [DOI] [PubMed] [Google Scholar]
  37. Moore L. K., Beyer E. C., Burt J. M. Characterization of gap junction channels in A7r5 vascular smooth muscle cells. Am J Physiol. 1991 May;260(5 Pt 1):C975–C981. doi: 10.1152/ajpcell.1991.260.5.C975. [DOI] [PubMed] [Google Scholar]
  38. Moreno A. P., Fishman G. I., Spray D. C. Phosphorylation shifts unitary conductance and modifies voltage dependent kinetics of human connexin43 gap junction channels. Biophys J. 1992 Apr;62(1):51–53. doi: 10.1016/S0006-3495(92)81775-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Musil L. S., Cunningham B. A., Edelman G. M., Goodenough D. A. Differential phosphorylation of the gap junction protein connexin43 in junctional communication-competent and -deficient cell lines. J Cell Biol. 1990 Nov;111(5 Pt 1):2077–2088. doi: 10.1083/jcb.111.5.2077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nishi M., Kumar N. M., Gilula N. B. Developmental regulation of gap junction gene expression during mouse embryonic development. Dev Biol. 1991 Jul;146(1):117–130. doi: 10.1016/0012-1606(91)90452-9. [DOI] [PubMed] [Google Scholar]
  41. Paul D. L., Ebihara L., Takemoto L. J., Swenson K. I., Goodenough D. A. Connexin46, a novel lens gap junction protein, induces voltage-gated currents in nonjunctional plasma membrane of Xenopus oocytes. J Cell Biol. 1991 Nov;115(4):1077–1089. doi: 10.1083/jcb.115.4.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Paul D. L. Molecular cloning of cDNA for rat liver gap junction protein. J Cell Biol. 1986 Jul;103(1):123–134. doi: 10.1083/jcb.103.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pepper M. S., Montesano R., el Aoumari A., Gros D., Orci L., Meda P. Coupling and connexin 43 expression in microvascular and large vessel endothelial cells. Am J Physiol. 1992 May;262(5 Pt 1):C1246–C1257. doi: 10.1152/ajpcell.1992.262.5.C1246. [DOI] [PubMed] [Google Scholar]
  44. Prentki M., Matschinsky F. M. Ca2+, cAMP, and phospholipid-derived messengers in coupling mechanisms of insulin secretion. Physiol Rev. 1987 Oct;67(4):1185–1248. doi: 10.1152/physrev.1987.67.4.1185. [DOI] [PubMed] [Google Scholar]
  45. Risek B., Gilula N. B. Spatiotemporal expression of three gap junction gene products involved in fetomaternal communication during rat pregnancy. Development. 1991 Sep;113(1):165–181. doi: 10.1242/dev.113.1.165. [DOI] [PubMed] [Google Scholar]
  46. Risek B., Guthrie S., Kumar N., Gilula N. B. Modulation of gap junction transcript and protein expression during pregnancy in the rat. J Cell Biol. 1990 Feb;110(2):269–282. doi: 10.1083/jcb.110.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Rubin J. B., Verselis V. K., Bennett M. V., Bargiello T. A. Molecular analysis of voltage dependence of heterotypic gap junctions formed by connexins 26 and 32. Biophys J. 1992 Apr;62(1):183–195. doi: 10.1016/S0006-3495(92)81804-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Segal S. S., Bény J. L. Intracellular recording and dye transfer in arterioles during blood flow control. Am J Physiol. 1992 Jul;263(1 Pt 2):H1–H7. doi: 10.1152/ajpheart.1992.263.1.H1. [DOI] [PubMed] [Google Scholar]
  49. Serras F., Notenboom R. G., Van den Biggelaar J. A. Communication compartments in the post-trochal ectoderm of the mollusc Lymnaea stagnalis. Exp Cell Res. 1990 Apr;187(2):284–291. doi: 10.1016/0014-4827(90)90093-p. [DOI] [PubMed] [Google Scholar]
  50. Simionescu M., Simionescu N., Palade G. E. Segmental differentiations of cell junctions in the vascular endothelium. The microvasculature. J Cell Biol. 1975 Dec;67(3):863–885. doi: 10.1083/jcb.67.3.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Spray D. C., Harris A. L., Bennett M. V. Equilibrium properties of a voltage-dependent junctional conductance. J Gen Physiol. 1981 Jan;77(1):77–93. doi: 10.1085/jgp.77.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Stutenkemper R., Geisse S., Schwarz H. J., Look J., Traub O., Nicholson B. J., Willecke K. The hepatocyte-specific phenotype of murine liver cells correlates with high expression of connexin32 and connexin26 but very low expression of connexin43. Exp Cell Res. 1992 Jul;201(1):43–54. doi: 10.1016/0014-4827(92)90346-a. [DOI] [PubMed] [Google Scholar]
  53. Swenson K. I., Jordan J. R., Beyer E. C., Paul D. L. Formation of gap junctions by expression of connexins in Xenopus oocyte pairs. Cell. 1989 Apr 7;57(1):145–155. doi: 10.1016/0092-8674(89)90180-3. [DOI] [PubMed] [Google Scholar]
  54. Swenson K. I., Piwnica-Worms H., McNamee H., Paul D. L. Tyrosine phosphorylation of the gap junction protein connexin43 is required for the pp60v-src-induced inhibition of communication. Cell Regul. 1990 Dec;1(13):989–1002. doi: 10.1091/mbc.1.13.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Valdimarsson G., De Sousa P. A., Beyer E. C., Paul D. L., Kidder G. M. Zygotic expression of the connexin43 gene supplies subunits for gap junction assembly during mouse preimplantation development. Mol Reprod Dev. 1991 Sep;30(1):18–26. doi: 10.1002/mrd.1080300103. [DOI] [PubMed] [Google Scholar]
  56. Veenstra R. D. Voltage-dependent gating of gap junction channels in embryonic chick ventricular cell pairs. Am J Physiol. 1990 Apr;258(4 Pt 1):C662–C672. doi: 10.1152/ajpcell.1990.258.4.C662. [DOI] [PubMed] [Google Scholar]
  57. Wang H. Z., Li J., Lemanski L. F., Veenstra R. D. Gating of mammalian cardiac gap junction channels by transjunctional voltage. Biophys J. 1992 Jul;63(1):139–151. doi: 10.1016/S0006-3495(92)81573-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Werner R., Levine E., Rabadan-Diehl C., Dahl G. Formation of hybrid cell-cell channels. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5380–5384. doi: 10.1073/pnas.86.14.5380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Willecke K., Heynkes R., Dahl E., Stutenkemper R., Hennemann H., Jungbluth S., Suchyna T., Nicholson B. J. Mouse connexin37: cloning and functional expression of a gap junction gene highly expressed in lung. J Cell Biol. 1991 Sep;114(5):1049–1057. doi: 10.1083/jcb.114.5.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Zhang J. T., Nicholson B. J. Sequence and tissue distribution of a second protein of hepatic gap junctions, Cx26, as deduced from its cDNA. J Cell Biol. 1989 Dec;109(6 Pt 2):3391–3401. doi: 10.1083/jcb.109.6.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular Biology of the Cell are provided here courtesy of American Society for Cell Biology

RESOURCES