June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Wnt signaling pathway in the human limbus: a comprehensive mapping by single mRNA detection
Author Affiliations & Notes
  • Clemence Bonnet
    Cornea, Jules Stein Eye Institute, Los Angeles, California, United States
    Cornea, Hopital Cochin, Paris, Île-de-France, France
  • Maxime Ruiz
    Cornea, Jules Stein Eye Institute, Los Angeles, California, United States
  • Sheyla Gonzalez
    Cornea, Jules Stein Eye Institute, Los Angeles, California, United States
  • Chi-Hong Tseng
    Cornea, Jules Stein Eye Institute, Los Angeles, California, United States
  • Jean-Louis Bourges
    Cornea, Hopital Cochin, Paris, Île-de-France, France
  • Francine F Behar-Cohen
    Cornea, Hopital Cochin, Paris, Île-de-France, France
  • Sophie Xiaohui Deng
    Cornea, Jules Stein Eye Institute, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Clemence Bonnet None; Maxime Ruiz None; Sheyla Gonzalez None; Chi-Hong Tseng None; Jean-Louis Bourges None; Francine Behar-Cohen None; Sophie Deng None
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 825. doi:
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      Clemence Bonnet, Maxime Ruiz, Sheyla Gonzalez, Chi-Hong Tseng, Jean-Louis Bourges, Francine F Behar-Cohen, Sophie Xiaohui Deng; Wnt signaling pathway in the human limbus: a comprehensive mapping by single mRNA detection. Invest. Ophthalmol. Vis. Sci. 2022;63(7):825.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : To evaluate the in situ spatial mRNA expression of 4 Wnt ligands (WNT2, WNT6, WNT11, WNT16B), 4 Wnt inhibitors (dickkopf WNT Signaling Pathway Inhibitor 1 [DKK1], secreted frizzled-related protein 5 [SFRP5], frizzled-related protein [FRZB], WNT inhibitory factor 1 [WIF1]), and frizzled7 (Fzd7), that have been shown previously upregulated in the limbus by differential gene profiling using q-RT-PCR.

Methods : The V2 fluorescent RNAscope (ACD, Newark, California) was performed on fresh frozen tissue sections from 5 human corneas with less than 8 hours of death to preservation time and intact epithelium. Positive signals were quantified using Imaris software.

Results : All 4 Wnt ligands, 4 Wnt inhibitors, and Fzd7 were preferentially expressed in the basal layer of the cornea and limbus compared to the suprabasal layer (P<0.05). When compared with the basal corneal epithelial layer, all 4 Wnt ligands, FRZB, and Fzd7 were preferentially expressed in the basal limbal epithelial layer in a decreasing gradient manner towards the superficial layers (P<0.05). No gradients were found for DKK1, WIF1, and SFRP5 expression. All 4 Wnt ligands, 4 Wnt inhibitors, and Fzd7 were also detected in the limbal stroma (P<0.05). Fzd7 was preferentially expressed in the basal limbal epithelial layer of the superior limbus compared with other limbal quadrants (P<0.05).

Conclusions : Wnt ligands are expressed in a gradient manner towards the superficial layers. Both paracrine and autocrine Wnt signaling pathways are involved in LSCs regulation, suggesting that a fine tuning of Wnt signaling exists to control LSC differentiation and proliferation.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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