September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Intraluminal Microtubule Acetylation Regulates Ocular Development and Corneal Repair
Author Affiliations & Notes
  • Lining Cao
    Department of Ophthalmology, UCSF Medical School, San Francisco, CA, USA, San Francisco, California, United States
  • Zhenjie Xu
    Department of Anatomy, UCSF Medical School, San Francisco, California, United States
    Department of Dermatology, Stanford University School of Medicine, Stanford, California, United States
  • Bo Liu
    Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford School of Medicine, Stanford, CA, USA, Stanford, California, United States
  • Ying Yu
    Department of Anatomy, UCSF Medical School, San Francisco, California, United States
  • Marcel Alavi
    Department of Ophthalmology, UCSF Medical School, San Francisco, CA, USA, San Francisco, California, United States
  • Jill Helms
    Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford School of Medicine, Stanford, CA, USA, Stanford, California, United States
  • Matilda F Chan
    Department of Ophthalmology, UCSF Medical School, San Francisco, CA, USA, San Francisco, California, United States
    Francis I. Proctor Foundation, University of California, San Francisco, California, United States
  • Peter Marinkovich
    Department of Dermatology, Stanford University School of Medicine, Stanford, California, United States
  • Zena Werb
    Department of Anatomy, UCSF Medical School, San Francisco, California, United States
  • Footnotes
    Commercial Relationships   Lining Cao, None; Zhenjie Xu, None; Bo Liu, None; Ying Yu, None; Marcel Alavi, None; Jill Helms, None; Matilda Chan, None; Peter Marinkovich, None; Zena Werb, None
  • Footnotes
    Support  Research Evaluation and Allocation Committee (REAC) Award
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 1278. doi:
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      Lining Cao, Zhenjie Xu, Bo Liu, Ying Yu, Marcel Alavi, Jill Helms, Matilda F Chan, Peter Marinkovich, Zena Werb; Intraluminal Microtubule Acetylation Regulates Ocular Development and Corneal Repair. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1278.

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      © 2017 Association for Research in Vision and Ophthalmology.

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Abstract

Purpose : Microtubules (MTs) are dynamic polymers that have remained the subject of intense investigation for their roles in cell division, migration and polarity. Acetylated microtubules are present in virtually all ocular developmental processes but their roles in normal ocular physiology are unknown. Our study examined the role of tubulin acetyltransferase, Mec17, in ocular development and corneal repair.

Methods : Mec17-/- mice were generated by replacing a 9.2 kB fragment of the Mec17 gene with a LacZ-neomycin reporter. Whole eyes of littermate WT and Mec17-/- mice at age E14.5 were processed for H&E staining. Mec17 expression was detected by β-galactosidase staining and acetylated a-tubulin (Ac-MT) and ZO-1 expression were analyzed by immunofluorescence staining. Corneal epithelial and endothelial injuries were induced by mechanical (scratch) and chemical injury (alkali) respectively.

Results : Mec17-/- mice at E14.5 demonstrate abnormal development of major ocular structures including the cornea, lens, retina, and optic nerve with incomplete penetrance. Wild-type adult mice express Mec17 and in the cornea (all layers), retina (GCL, OPN and IS/OS layers), and optic nerve. Acetylated alpha-tubulin is expressed in the retina (GCL, IPL, OPL and IS/OS layers) and optic nerve. Following corneal epithelial injury, Mec17-/- mice showed delayed epithelial closure at 8 and 24 hours post-injury compared with littermate WT mice. Following corneal alkali injury, WT corneal endothelium displayed regular ZO-1 staining and dot-like acetylated a-Tubulin, but Mec17-/- corneas displayed disorganized ZO-1 staining and lacked acetylated a-Tubulin staining.

Conclusions : Mec17 has an important role in normal ocular development and is highly expressed in the adult cornea, retina, and optic nerve. Lack of Mec17 expression results in abnormal corneal epithelial and endothelial repair. These results establish an essential function of Mec17 in normal ocular developmental and physiologic processes.

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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