September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Cell-Cell Adhesion Mediated by the Second Extracellular Domain of Connexin 50 Promotes Lens Epithelial-Fiber Cell Differentiation
Author Affiliations & Notes
  • Jean X Jiang
    Biochemistry, University of Texas Health Science Center, San Antonio, Texas, United States
  • Zhengping Hu
    Biochemistry, University of Texas Health Science Center, San Antonio, Texas, United States
  • Wen Shi
    Biochemistry, University of Texas Health Science Center, San Antonio, Texas, United States
  • Sumin Gu
    Biochemistry, University of Texas Health Science Center, San Antonio, Texas, United States
  • Footnotes
    Commercial Relationships   Jean Jiang, None; Zhengping Hu, None; Wen Shi, None; Sumin Gu, None
  • Footnotes
    Support  NIH Grant EY012085
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 3070. doi:
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      Jean X Jiang, Zhengping Hu, Wen Shi, Sumin Gu; Cell-Cell Adhesion Mediated by the Second Extracellular Domain of Connexin 50 Promotes Lens Epithelial-Fiber Cell Differentiation. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3070.

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      © 2017 Association for Research in Vision and Ophthalmology.

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Abstract

Purpose : We have recently shown that connexin (Cx) 50 acts as a cell adhesion molecule in lens fiber cells and enhances the cell adhesion mediated by AQP0. However, the impact and molecular mechanism underlying this adhesion function of Cx50 in the lens fiber remains elusive.

Methods : High titer recombinant retroviruses containing vehicle and chick Cx50 were prepared and used to infect chick embryonic fibroblast (CEF) cells. Cell-cell adhesion assay was conducted using a “parachuting” adhesion method with fluorescence-labeled donor or receipt cells expressing Cx50 or vehicle controls. Lens primary cell cultures were prepared using embryonic day-11 fertilized chicken eggs. GST fusion protein containing the first (E1) or second (E2) extracellular domain of Cx50 was generated and was added to lens primary cell culture every two days. The extent of lens epithelial-fiber differentiation was determined by counting numbers of lentoids and the expression of AQP0 protein detected by immunofluorescence using anti-Cx50 antibody.

Results : We showed that the expression of Cx50 promoted cell adhesion at a comparable level to AQP0; however, this increase was not observed with other lens connexins, Cx43 and Cx46. Moreover, this adhesive property occurred in both homotypic with Cx50 expressed in both pairing cells as well as heterotypic with Cx50 in only one pairing cell. Treatment with a fusion protein targeting the E2 domain of Cx50 inhibited cell adhesion and such inhibitory effect was not observed with the E1 domain. These data suggest that the adhesive function is mediated by the E2 domain. Furthermore, disruption of cell adhesion by incubation with the E2 domain also impaired primary lens cell differentiation indicated by reduced lentoids formation and AQP0 expression.

Conclusions : These results suggest that in addition to its role in forming gap junction channels, Cx50 plays a unique role in mediating cell-cell adhesion function in the lens. The extracellular E2 domain is involved in this adhesive role of Cx50. Moreover, the cell adhesion mediated by Cx50 plays a critical role in lens epithelial-fiber cell differentiation.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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