June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
REMODELING OF CORNEAL COLD SENSORY NERVE FIBERS IN THE ADULT LIVING MOUSE
Author Affiliations & Notes
  • Almudena Íñigo-Portugués
    Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, San Juan de Alicante, Alicante, Spain
  • Giovanna Exposito
    Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, San Juan de Alicante, Alicante, Spain
  • Juana Gallar
    Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, San Juan de Alicante, Alicante, Spain
  • Carlos Belmonte
    Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, San Juan de Alicante, Alicante, Spain
  • Victor Meseguer
    Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, San Juan de Alicante, Alicante, Spain
  • Footnotes
    Commercial Relationships   Almudena Íñigo-Portugués, None; Giovanna Exposito, None; Juana Gallar, None; Carlos Belmonte, None; Victor Meseguer, None
  • Footnotes
    Support  SAF2014-54518-C3-1-R; SAF2014-54518-C3-2-R
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1021. doi:
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      Almudena Íñigo-Portugués, Giovanna Exposito, Juana Gallar, Carlos Belmonte, Victor Meseguer; REMODELING OF CORNEAL COLD SENSORY NERVE FIBERS IN THE ADULT LIVING MOUSE. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1021.

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

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Abstract

Purpose : To define the time course and magnitude of remodeling of corneal cold sensory nerve fibers of adult mice in vivo.

Methods : Mice expressing EYFP under the control of TRPM8 regulatory sequences (TRPM8-EYFP mouse) were anesthetized with an intraperitoneal injection of xylazine (16 mg/kg) followed by inhalation of isoflurane (1.5 %). In TRPM8-EYFP mice, only cold-sensitive fibers (roughly 20 percent of the total) display a green fluorescence, thus allowing their morphological identification. TRPM8+ sub-basal nerve leashes and intraepithelial terminals were imaged at 5x and 25x magnification by means of a confocal microscope and reconstructed in 3D using Imaris 8.2 software. Individual corneal leashes and intraepithelial terminals were monitored in the same animal at different time points over a week (day 0, day 2 and day 7). The total length of individual leashes was measured, and the length change rate calculated. The number of nerve terminals given by each sub-basal nerve fiber was also counted.

Results : Points of penetration of a single stromal nerve into basal lamina to form a sub-basal leash were not homogeneously distributed throughout the cornea, being more abundant in its periphery than in the center. Five out of 7 leashes containing TRPM8 axons, measured in 3 mice, increased their total length at a rate of 34.56 ± 14.10 µm/day. Contrarily, total length decreased in another 2 leashes at a rate of -55.38 ± 34.60 µm/day. Additionally, the number, location and shape of nerve terminals given by the leashes changed with time (figure 1).

Conclusions : Corneal nerve fibers in the living animal experience dynamic configuration changes involving increases and decreases of leashes’ total length and redistribution of their terminal endings, reflecting a continuous remodeling of corneal epithelium innervation.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

 

Remodeling of TRPM8+ leashes and intraepithelial terminals in living mice. (A) Confocal micrograph of a subbasal leash and its nerve terminals in 3-D taken at day 0, and (B) at day 7. Asterisk indicates the reference point in the subepithelial plexus used in both images. Rendering in 3-D at day 0 (C) and day 7 (D) of the same sub-basal leash (red) and nerve terminals (in gray). Scale bar: 70 mm

Remodeling of TRPM8+ leashes and intraepithelial terminals in living mice. (A) Confocal micrograph of a subbasal leash and its nerve terminals in 3-D taken at day 0, and (B) at day 7. Asterisk indicates the reference point in the subepithelial plexus used in both images. Rendering in 3-D at day 0 (C) and day 7 (D) of the same sub-basal leash (red) and nerve terminals (in gray). Scale bar: 70 mm

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