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. 1970 May 1;45(2):272–290. doi: 10.1083/jcb.45.2.272

A FINE STRUCTURAL ANALYSIS OF INTERCELLULAR JUNCTIONS IN THE MOUSE LIVER

Daniel A Goodenough 1, Jean Paul Revel 1
PMCID: PMC2107902  PMID: 4105112

Abstract

Zonulae occludentes and gap junctions were examined both in the intact mouse liver and in a junction-rich membrane fraction from homogenized mouse liver. These preparations were visualized with the techniques of uranyl acetate staining en bloc, staining with colloidal lanthanum, negative staining with phosphotungstate, and freeze-cleaving. The zonula occludens is arranged as a meshwork of branching and anastomosing threadlike contacts sealing the lumen of the bile canaliculus from the liver intercellular space. The gap junction is characterized in section by a 20 A gap between the apposed junctional membrane outer leaflets, and permeation of this space with lanthanum or phosphotungstate reveals a polygonal lattice of subunits with a center-to-center spacing of 90–100 A. Freeze-cleaved gap junctions show a similar lattice. Extraction of junction-rich fractions with 60% aqueous acetone results in a disappearance of the 20 A gap in sectioned pellets and an inability to demonstrate the polygonal lattice with either the freeze-cleave or negative staining techniques. Extraction of the membranes with 50% acetone does not produce this effect. Thin-layer chromatography of the acetone extracts reveals a group of phospholipids in the 60% extract that are not detectable in the 50% extract. Acetone does not cause any detectable change in the structure of the zonula occludens, but the occluding junction becomes leaky to lanthanum following acetone treatment. The effects of other reagents on the junctions are reported.

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Selected References

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  1. BARR L., DEWEY M. M., BERGER W. PROPAGATION OF ACTION POTENTIALS AND THE STRUCTURE OF THE NEXUS IN CARDIAC MUSCLE. J Gen Physiol. 1965 May;48:797–823. doi: 10.1085/jgp.48.5.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BENNETT H. S., LUFT J. H. zeta-Collidine as a basis for buffering fixatives. J Biophys Biochem Cytol. 1959 Aug;6(1):113–114. doi: 10.1083/jcb.6.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benedetti E. L., Emmelot P. Electron microscopic observations on negatively stained plasma membranes isolated from rat liver. J Cell Biol. 1965 Jul;26(1):299–305. doi: 10.1083/jcb.26.1.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennett M. R., Rogers D. C. A study of the innervation of the taenia coli. J Cell Biol. 1967 Jun;33(3):573–596. doi: 10.1083/jcb.33.3.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Branton D., Park R. B. Subunits in chloroplast lamellae. J Ultrastruct Res. 1967 Aug;19(3):283–303. doi: 10.1016/s0022-5320(67)80222-3. [DOI] [PubMed] [Google Scholar]
  6. Brightman M. W., Reese T. S. Junctions between intimately apposed cell membranes in the vertebrate brain. J Cell Biol. 1969 Mar;40(3):648–677. doi: 10.1083/jcb.40.3.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bullivant S., Ames A., 3rd A simple freeze-fracture replication method for electron microscopy. J Cell Biol. 1966 Jun;29(3):435–447. doi: 10.1083/jcb.29.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bullivant S., Loewenstein W. R. Structure of coupled and uncoupled cell junctions. J Cell Biol. 1968 Jun;37(3):621–632. doi: 10.1083/jcb.37.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DEWEY M. M., BARR L. A STUDY OF THE STRUCTURE AND DISTRIBUTION OF THE NEXUS. J Cell Biol. 1964 Dec;23:553–585. doi: 10.1083/jcb.23.3.553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dewey M. M., Barr L. Intercellular Connection between Smooth Muscle Cells: the Nexus. Science. 1962 Aug 31;137(3531):670–672. doi: 10.1126/science.137.3531.670-a. [DOI] [PubMed] [Google Scholar]
  11. Dod B. J., Gray G. M. The lipid composition of rat-liver plasma membranes. Biochim Biophys Acta. 1968 Apr 29;150(3):397–404. doi: 10.1016/0005-2736(68)90138-7. [DOI] [PubMed] [Google Scholar]
  12. EMMELOT P., BOS C. J., BENEDETTI E. L., RUEMKE P. STUDIES ON PLASMA MEMBRANES. I. CHEMICAL COMPOSITION AND ENZYME CONTENT OF PLASMA MEMBRANES ISOLATED FROM RAT LIVER. Biochim Biophys Acta. 1964 Jul 15;90:126–145. doi: 10.1016/0304-4165(64)90125-4. [DOI] [PubMed] [Google Scholar]
  13. FARQUHAR M. G., PALADE G. E. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. doi: 10.1083/jcb.17.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. FURSHPAN E. J. "ELECTRICAL TRANSMISSION" AT AN EXCITATORY SYNAPSE IN A VERTEBRATE BRAIN. Science. 1964 May 15;144(3620):878–880. doi: 10.1126/science.144.3620.878. [DOI] [PubMed] [Google Scholar]
  15. Farquhar M. G., Palade G. E. Cell junctions in amphibian skin. J Cell Biol. 1965 Jul;26(1):263–291. doi: 10.1083/jcb.26.1.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  17. Heath T., Wissig S. L. Fine structure of the surface of mouse hepatic cells. Am J Anat. 1966 Jul;119(1):97–127. doi: 10.1002/aja.1001190107. [DOI] [PubMed] [Google Scholar]
  18. Karnovsky M. J. The ultrastructural basis of capillary permeability studied with peroxidase as a tracer. J Cell Biol. 1967 Oct;35(1):213–236. doi: 10.1083/jcb.35.1.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koehler J. K. The technique and application of freeze-etching in ultrastructure research. Adv Biol Med Phys. 1968;12:1–84. doi: 10.1016/b978-1-4831-9928-3.50005-9. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Loewenstein W. R. Permeability of membrane junctions. Ann N Y Acad Sci. 1966 Jul 14;137(2):441–472. doi: 10.1111/j.1749-6632.1966.tb50175.x. [DOI] [PubMed] [Google Scholar]
  22. MILLER F. Hemoglobin absorption by the cells of the proximal convoluted tubule in mouse kidney. J Biophys Biochem Cytol. 1960 Dec;8:689–718. doi: 10.1083/jcb.8.3.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. MOLLENHAUER H. H. PLASTIC EMBEDDING MIXTURES FOR USE IN ELECTRON MICROSCOPY. Stain Technol. 1964 Mar;39:111–114. [PubMed] [Google Scholar]
  24. Muir A. R. The effects of divalent cations on the ultrastructure of the perfused rat heart. J Anat. 1967 Apr;101(Pt 2):239–261. [PMC free article] [PubMed] [Google Scholar]
  25. NEVILLE D. M., Jr The isolation of a cell membrane fraction from rat liver. J Biophys Biochem Cytol. 1960 Oct;8:413–422. doi: 10.1083/jcb.8.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Penn R. D. Ionic communication between liver cells. J Cell Biol. 1966 Apr;29(1):171–174. doi: 10.1083/jcb.29.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. ROBERTSON J. D. THE OCCURRENCE OF A SUBUNIT PATTERN IN THE UNIT MEMBRANES OF CLUB ENDINGS IN MAUTHNER CELL SYNAPSES IN GOLDFISH BRAINS. J Cell Biol. 1963 Oct;19:201–221. doi: 10.1083/jcb.19.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. ROUSER G., KRITCHEVSKY G., GALLI C., HELLER D. DETERMINATION OF POLAR LIPIDS: QUANTITATIVE COLUMN AND THIN-LAYER CHROMATOGRAPHY. J Am Oil Chem Soc. 1965 Mar;42:215–227. doi: 10.1007/BF02541135. [DOI] [PubMed] [Google Scholar]
  29. Reese T. S., Karnovsky M. J. Fine structural localization of a blood-brain barrier to exogenous peroxidase. J Cell Biol. 1967 Jul;34(1):207–217. doi: 10.1083/jcb.34.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Revel J. P., Hamilton D. W. The double nature of the intermediate dense line in peripheral nerve myelin. Anat Rec. 1969 Jan;163(1):7–15. doi: 10.1002/ar.1091630102. [DOI] [PubMed] [Google Scholar]
  31. Revel J. P., Karnovsky M. J. Hexagonal array of subunits in intercellular junctions of the mouse heart and liver. J Cell Biol. 1967 Jun;33(3):C7–C12. doi: 10.1083/jcb.33.3.c7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sheridan J. D. Electrophysiological evidence for low-resistance intercellular junctions in the early chick embryo. J Cell Biol. 1968 Jun;37(3):650–659. doi: 10.1083/jcb.37.3.650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sheridan J. D. Electrophysiological study of special connections between cells in the early chick embryo. J Cell Biol. 1966 Oct;31(1):C1–C5. doi: 10.1083/jcb.31.1.c1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Skipski V. P., Barclay M., Archibald F. M., Terebus-Kekish O., Reichman E. S., Good J. J. Lipid composition of rat liver cell membranes. Life Sci. 1965 Sep;4(17):1673–1680. doi: 10.1016/0024-3205(65)90213-4. [DOI] [PubMed] [Google Scholar]
  35. Skipski V. P., Smolowe A. F., Sullivan R. C., Barclay M. Separation of lipid classes by thin-layer chromatography. Biochim Biophys Acta. 1965 Oct 4;106(2):386–396. doi: 10.1016/0005-2760(65)90047-0. [DOI] [PubMed] [Google Scholar]
  36. Staehelin L. A., Mukherjee T. M., Williams A. W. Freeze-etch appearance of the tight junctions in the epithelium of small and large intestine of mice. Protoplasma. 1969;67(2):165–184. doi: 10.1007/BF01248737. [DOI] [PubMed] [Google Scholar]
  37. Trelstad R. L., Hay E. D., Revel J. D. Cell contact during early morphogenesis in the chick embryo. Dev Biol. 1967 Jul;16(1):78–106. doi: 10.1016/0012-1606(67)90018-8. [DOI] [PubMed] [Google Scholar]
  38. WAGNER H., HOERHAMMER L., WOLFF P. [Thin layer chromatography of phosphatides and glycolipids]. Biochem Z. 1961;334:175–184. [PubMed] [Google Scholar]

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