2011 Group 3 Project

From CellBiology

Tight Junctions

--Mark Hill 15:32, 2 May 2011 (EST)--Mark Hill 14:01, 28 April 2011 (EST) R.D and E.C where are your contributions? The "bullet point" listing is also not the easiest form to get all your information across. No images on the page?


Tight junctions, or zona occludens, are a branching network of sealing proteins found on the apical ends of epithelia. Tight junctions play significant roles in the integrity of the cell as well as cell-cell transport through the formation of a barrier for diffusion. In vertebrates, tight junctions are associated with the junctional complex. Analogous forms of tight junctions are found in invertebrates in the form of septate junctions. (SUBJECT TO CHANGE)


Early 1940’s: Hans Ussing and associates propose the “two membrane model” to address the question of how sodium ions are moved in a directional fashion across the epithelium. This model stated that the apical and basal membrane surfaces have different conductance properties, namely that sodium enters the cell across the apical membrane down its concentration gradient. [1]

1963: Farquhar and Palade first visualizes the tight junctions at the ultrastructural level. [2]

1964: Hans Ussing and associates discover inconsistencies within their findings leading to the understanding that Chloride ions follow sodium ions in a passive fashion through the tight junction which they called the “shunt pathway”. [3]

Gradually, it was accepted that intact normal epithelia display a wide range of electrical resistances and that it was the tight junctions that determined whether the transepithelial resistance is high or low[4]

1986: Stevenson and associates isolate and localize a component of the cytoplasmic face of the tight junction naming the protein Zona occludens-1 (ZO-1). [5]

1988-1991: Citi and associates discover and localise major tight junction plaque protein cingulin. [6][7][8]

1991: Gumbiner and associates identify the tight junction protein ZO-2.[9]

1993: Furuse and associates immunolocalised a tetraspan transmembrane protein of the Zona Occludens naming it "occludin".[10]

1994-1997: Furuse and associates transfect "occludin" encoding cDNA. Transfection of cDNA and expression caused the construction of tight junction possessing cells with occludin integrated into pre-existing tight junction structures. When injected into tight junction lacking cells, it was able to form de novo tight junction typical intramembranous “ridges” as well as cytoplasmic stacks of lamellae resembling tight junction membrane structures. [11][12]

1998: Furuse and associates identify 2 smaller polypeptides. Immunolocalisation and incorporation into tight junctions of living cells formed de novo intramembranous TJ typical ridge structures, even in TJ lacking cells naming these proteins as “claudins” nos. 1 and 2. [13][14]

1998: Haskins and associates identify the tight junction protein ZO-3. [15]

2001-2002: Tsukita and associates discovery a total of 24 different claudin proteins encoded in the human genome.[16][17]

2002: Bradner & Furuse and associates observe typical tight junction structures, positive for claudins and occluding in the uppermost layers of the epidermis and other squamous stratified epithelia. [18][19]

2005: Ikenouchi & associates and Tsukita & associates– Discover small intercellular “gaps” formed by three cells meeting together posing a “three corner state problem” (Ikenouchi). The answer to this problem came in the form of a protein, which Tsukita named “tricellulin”. This protein was found to assemble exactly at those “three corner” sites. [20]


Schematic model of the basic structural transmembrane components of tight junctions

Tight junctions consist of a branching network of sealing strands of protein. Each strand is assembled from a series of transmembrane proteins(JAMs/Junctional Adhesion Molecules,Claudins and Occludin) embedded in plasma membranes. The extracellular domains join each other in tight junctions,whereas the intracellular domains are linked to peripheral membrane proteins,linking the transmembrane protein strands to actin cytoskeleton to create a functional network that plays a role in cellular processes. Each strand functions as an individual or linear barrie, therefore, the number of transmembrane protein strands is in relation with the degree of paracellular electrical resistance and impedance to solute flux in tight junctions[21]. Although other types of proteins are present at tight junctions, however, occludin and claudin are the major ones contributing to the structure of tight junctions.

Links to other images of tight junctions

Model of tight junctions

Electron microscope image of tight junction

Molecular components

Associated molecular components of tight junctions,

Two groups of proteins:

  • Transmembrane proteins
  • Peripheral membrane proteins

Transmembrane proteins serve as important components of the tight junction that span across the junction, connecting two adjacent cells and making a seal tight at the junction.The group has three members.

  • Occludins
  • Claudins
  • Junctional Adhesion Molecules (JAMs)


  • family of transmembrane proteins.
  • the first tight junction associated transmembrane proteins and were identified in chicken liver[22].
  • consists of four transmembrane domains,two extracellular loops and two intracellular domains[23].


  • interacting directly with proteins zonula occludens,ZO-1 and ZO-2 and ZO-3 at the tight junction to localize occludins
  • interacting indirectly with the actin cytoskeleton and junctional adhesion molecules(JAMs) via interacting with ZO proteins[24]
  • involving in cell-cell adhesion with extracellular domains of occludin[25]
  • vital in tight junction assembly in Xenopus embryo development[26]
  • regulating various signaling events initiating from the tight junction[27]
  • may involve in RhoA activation via a tight junction–associated guanine nucleotide exchange factor,GEF-H1/Lfc[28][29]
  • targeting TGF-ß receptors to tight junctions[30].


  • family of transmembrane proteins that are also the main constituents of the tight junction intercellular strands
  • claudin-1 and claudin-2 were the first claudin family members identified in a chicken liver fraction[31]
  • important for the barrier function of tight junctions[32].


  • forming tight junction strands via interacting with each other between different tight junction strands or within individual strands in a homotypic and heterotypic manner [33]
  • structural components are similar to occuldin.Four transmembrane domains, two extracellular loops, and two intracellular domains,however,it does not have sequence similarity to occludin [34]
  • mediate calcium-independent cell-cell adhesion[35]
  • interacting directly with peripheral PDZ-domain-containing proteins(ZO-1, ZO-2, ZO-3,and protein associated with Lin seven 1

(PALS1)-associated tight junction protein (PATJ))[36]

Junctional adhesion molecules (JAMs)

  • immunoglobulin superfamily of proteins and are expressed in epithelial cells,leukocytes, endothelia, and platelets
  • four members of the JAM protein family have been identified so far, JAM-A, JAM-B, JAM-C, and JAM4/JAML[37]
  • in epithelia, JAMs are directed to tight junctions, whereas,JAM-B exists along the lateral membrane[38]
  • have a single transmembrane domain, an extracellular domain containing two Ig-like motifs, and a cytoplasmic tail[39].


  • may participate in cell adhesion via homophilic interactions[40]
  • interacting with each other. JAM-B interacts with JAM-C and integrins through heterophilic interactions[41]
  • forming intercellular junctions and epithelial barrier function[42].

Peripheral membrane proteins serve as intracellular binding sites for transmembrane proteins to allow transmembrane proteins to be organized in membrane and attached to the cytoskeleton to initiate cell signaling.The group has four members.

  • Zonula occludens(ZOs)
  • Membrane-associated guanylate kinase inverted(MAGI)
  • Cingulin
  • Rab13

Zonula occludens(ZOs)

  • tight junction proteins in the membrane-associated guanylate kinase (MAGUK) family of proteins
  • contain a core structure consisting of one or more PDZ domains, an Src homology 3 (SH3) domain, and a guanylate kinase (GUK) domain
  • three isoforms of ZO proteins: ZO-1, ZO-2, and ZO-3[43].


  • serving as peripheral membrane scaffolding proteins
  • interacting with many binding partners at the tight junction
  • involving in the formation of tight junction[44].

Membrane-associated guanylate kinase inverted(MAGI)

  • MAGUK family of proteins
  • three features that distinguish them from all other members of the family:

(a) The GUK domain is located on the N terminus

(b) the SH3 domain is replaced by two WW domains and

(c) MAGI proteins contain five PDZ domains[45]


  • associating with ß-catenin in E-cadherin-based adherens junctions
  • involving in the formation of adherens and tight junctions
  • involving in the interaction involving junctional proteins during the polarization process[46]
  • playing role in actin-cytoskeleton dynamics within polarized epithelial cells[47]
  • involving in various signal transduction events[48]


  • a 140–160-kD protein localizes to the cytoplasmic surface of epithelial tight junctions
  • was originally identified as a peripheral membrane protein at the tight junction in avian brush border cells[49]
  • has globular head and tail domains as well as a central a-helical rod domain[50].


  • potential role in embryogenesis and epithelial maturation[51]
  • linking tight junction proteins to the actin cytoskeleton[52]
  • involving in transcriptional regulation and cell proliferation[53]
  • playing a role in cell-cycle progression[54].

ZO-1-associated nucleic acid–binding protein(ZONAB)

  • was initially identified as a binding partner of ZO-1
  • a Y-box transcription factor protein [55].


  • interacting with cell division kinase 4 (CDK4) to regulate cell proliferation
  • serving as a sensor of cell density[56].


  • member of the small GTPase Rab family of proteins
  • was first identified as a mammalian homolog of the yeast secretory protein, Sec4[57]


  • involving in the regulation of exocytic and endocytic pathways, including vesicle movement and fusion[58]
  • involving in early junctional formation[59] .



Transepithelial transport




Current associated research




Important applications



Further Information

Reference list

Coordinator Comment to all Groups

I will add a general comment that will be the same to all groups under this heading.

Referencing Extension Problem

--Mark Hill 13:16, 3 May 2011 (EST) As mentioned in the lecture, I am aware of the referencing extension problem on your project pages. I have the following temporary solution, of removing the extension, so that groups can continue to add content to their project pages. I am also giving everyone a 1 week extension before the peer assessment.

This should only be done if your project page is not allowing you to save changes!

A. The Easy Way....

The following 4 steps can be done on the webpage or select all content in edit mode, copy and paste into a text editor. All steps must be completed before you attempt to save.

  1. In page edit mode, find all <pubmed> reference tags.
  2. Replace this tag with [http://www.ncbi.nlm.nih.gov/pubmed/ Note, there should be no spaces between the internet address and the pmid number.
  3. Now find all </pubmed> reference tags.
  4. Replace this second tag with ]

This will generate a numbered reference list that we can later fix up.

B. The Better Looking Result....

Whatever is between the <ref> </ref></pubmed> tags is what will appear in your reference list, so you can format the reference and link to appear in your reference list.

2011 Projects: Synaptic Junctions | Gap Junctions | Tight Junctions | Desmosomes | Adherens Junctions | Neuromuscular Junction
  1. <pubmed>13544986</pubmed>
  2. <pubmed>13944428</pubmed>
  3. <pubmed>14209264</pubmed>
  4. <pubmed>20066090</pubmed>
  5. <pubmed>3528172</pubmed>
  6. <pubmed>3285223</pubmed>
  7. <pubmed>2693465</pubmed>
  8. <pubmed>2012170</pubmed>
  9. <pubmed>2014265</pubmed>
  10. <pubmed>8276896</pubmed>
  11. <pubmed>8838666</pubmed>
  12. <pubmed>7798316</pubmed>
  13. <pubmed>9647647</pubmed>
  14. <pubmed>9786950</pubmed>
  15. <pubmed>9531559</pubmed>
  16. <pubmed>10747082</pubmed>
  17. <pubmed>11283726</pubmed>
  18. <pubmed>12067061</pubmed>
  19. <pubmed>11889141</pubmed>
  20. <pubmed>16365161</pubmed>
  21. <pubmed>641977</pubmed>
  22. <pubmed>8276896</pubmed>
  23. <pubmed>15820558</pubmed>
  24. <pubmed>10966866</pubmed>
  25. <pubmed>9175707</pubmed>
  26. <pubmed>9265654</pubmed>
  27. <pubmed>15806147</pubmed>
  28. <pubmed>15866167</pubmed>
  29. <pubmed>12604587</pubmed>
  30. <pubmed>15761153</pubmed>
  31. <pubmed>9647647</pubmed>
  32. <pubmed>16771626</pubmed>
  33. <pubmed>10562289</pubmed>
  34. <pubmed>16771626</pubmed>
  35. <pubmed>9786950</pubmed>
  36. <pubmed>11283726</pubmed>
  37. <pubmed>15820556</pubmed>
  38. <pubmed>11739175</pubmed>
  39. <pubmed>11500366</pubmed>
  40. <pubmed>10913139</pubmed>
  41. <pubmed>12070135</pubmed>
  42. <pubmed>10852816</pubmed>
  43. <pubmed>10966866</pubmed>
  44. <pubmed>12482754</pubmed>
  45. <pubmed>9395497</pubmed>
  46. <pubmed>10772923 </pubmed>
  47. <pubmed>12042308</pubmed>
  48. <pubmed>15579911</pubmed>
  49. <pubmed>3285223</pubmed>
  50. <pubmed>11042084</pubmed>
  51. <pubmed>8325238</pubmed>
  52. <pubmed>12023291</pubmed>
  53. <pubmed>15454572</pubmed>
  54. <pubmed>15866167</pubmed>
  55. <pubmed>10790369</pubmed>
  56. <pubmed>12566432</pubmed>
  57. <pubmed>8294494</pubmed>
  58. <pubmed>8294494</pubmed>
  59. <pubmed>11025210</pubmed>