September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Regeneration and Healing of Corneal Stromal Nerve Trunks after In Situ Laser Transection.
Author Affiliations & Notes
  • Joy Sarkar
    Ophthalmology, Illinois Eye & Ear Infirmary, UIC, Chicago, Illinois, United States
  • Eunjae Kim
    Ophthalmology, Illinois Eye & Ear Infirmary, UIC, Chicago, Illinois, United States
  • Disha Varma
    Ophthalmology, Illinois Eye & Ear Infirmary, UIC, Chicago, Illinois, United States
  • Behrad Yousefi Milani
    Ophthalmology, Illinois Eye & Ear Infirmary, UIC, Chicago, Illinois, United States
  • Sandeep Jain
    Ophthalmology, Illinois Eye & Ear Infirmary, UIC, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Joy Sarkar, None; Eunjae Kim, None; Disha Varma, None; Behrad Milani, None; Sandeep Jain, None
  • Footnotes
    Support  NIH EY023656 (SJ) and RPB Grant (UIC)
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4916. doi:
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      Joy Sarkar, Eunjae Kim, Disha Varma, Behrad Yousefi Milani, Sandeep Jain; Regeneration and Healing of Corneal Stromal Nerve Trunks after In Situ Laser Transection.. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4916.

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

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Abstract

Purpose : We have previously reported that lamellar dissection of the cornea transects stromal nerves, and that regenerating neurites form a dense net along the surgical plane. In these experiments, we have disrupted the stromal nerve trunks in situ, without incising the cornea, to determine the regeneration events in the absence of a surgical plane.

Methods : Thy1-YFP mice were anesthetized and in vivo images of the corneal nerves were obtained with a wide-field stereofluorescent microscope. A far infrared Xyrcos Laser attached to 20X objective of an upright microscope was used to perform in situ transection of the stromal nerves. 3 types of laser transections were performed (n=5/group): (i) point transection (a single cut); (ii) segmental transection (two cuts enclosing a segment of nerve trunk); and (iii) annular transection (cuts on all nerve trunks crossing the perimeter of a 0.8 mm diameter circular area centered on the corneal apex). Mice were imaged sequentially for 4 weeks thereafter to assess nerve degeneration (disappearance or weakening of original fluorescence intensity) or regeneration (appearance of new fluorescent fronds). Beta-3-tubulin immunostaining was performed on corneal wholemounts to demonstrate nerve disruption.

Results : The pattern of stromal nerves in corneas of the same mouse and in corneas of littermates was dissimilar. Two distinct patterns were observed, often within the same cornea: (i) interconnected trunks that spanned limbus to limbus,; or (ii) dichotomously branching trunks that terminate at the corneal apex . Point transections did not cause degeneration of proximal or distal segment in interconnected trunks, but resulted in degeneration of distal segment of branching trunks. In segmental transections, the nerve segment enclosed within the two laser cuts degenerated. Lack of beta-3 tubulin staining at transection site confirmed nerve transection. In interconnected trunks, at 4 weeks, a hyperfluorescent plaque filled the gap created by the transection. In annular transections, some nerve trunks degenerated, while others regained or retained fluorescence.

Conclusions : Interconnected stromal nerves in murine corneas do not degenerate after in situ point transection and show evidence of healing at the site of disruption. Presence or absence of a surgical plane influences corneal nerve regeneration after transection.

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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