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Axonal debris accumulates in corneal epithelial cells after intraepithelial corneal nerves are damaged: A focused Ion Beam Scanning Electron Microscopy (FIB-SEM) study.
Exp Eye Res. 2020 05; 194:107998.EE

Abstract

The intraepithelial corneal nerves (ICNs) that innervate the corneal epithelium are maintained through interactions with corneal epithelial cells and the extracellular matrix they produce. One to several axons bundle together within the basal cell layer and extend parallel to the ocular surface or branch and extend apically. Here we use 3-dimentional (3D) ultrastructural reconstructions of control and trephine injured mouse corneal epithelium and stroma produced using Focused Ion Beam Scanning Electron Microscope (FIB-SEM) to determine whether corneal epithelial or immune cells resident in the epithelium remove axonal debris and degrade it in their lysosomes after trephine injury to the cornea. We demonstrate that axonal fragments are internalized in the corneal epithelium and accumulate within electron dense structures consistent with lysosomes 3 h after trephine injury in both epithelial and immune cells located among the basal cells of the trephine injured cornea. Confocal imaging showed fewer CD45[+] immune cells within the corneal epithelium after trephine injury compared to controls. The resolution obtained using FIB-SEM also allowed us to show that the presence of sensory axons at the basal aspect of the epithelial basal cells close to the anterior aspect of the epithelial basement membrane (EBM) is associated with a focal reduction in EBM thickness. In addition, we show using FIB-SEM and confocal imaging that superficial trephine injuries that do not penetrate the stroma, damage the integrity of anterior stromal nerves. These studies are the first to look at the mouse cornea following nerve injury using FIB-SEM.

Authors+Show Affiliations

GW Nanofabrication and Imaging Center, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA.Department of Anatomy and Cell Biology, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA.Department of Anatomy and Cell Biology, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA.Department of Anatomy and Cell Biology, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA.GW Nanofabrication and Imaging Center, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA; Department of Anatomy and Cell Biology, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA.Department of Anatomy and Cell Biology, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA; Department of Ophthalmology, The George Washington School of Medicine and Health Sciences, Washington DC, 20052, USA. Electronic address: mastepp@gwu.edu.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32209319

Citation

Parlanti, Paola, et al. "Axonal Debris Accumulates in Corneal Epithelial Cells After Intraepithelial Corneal Nerves Are Damaged: a Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) Study." Experimental Eye Research, vol. 194, 2020, p. 107998.
Parlanti P, Pal-Ghosh S, Williams A, et al. Axonal debris accumulates in corneal epithelial cells after intraepithelial corneal nerves are damaged: A focused Ion Beam Scanning Electron Microscopy (FIB-SEM) study. Exp Eye Res. 2020;194:107998.
Parlanti, P., Pal-Ghosh, S., Williams, A., Tadvalkar, G., Popratiloff, A., & Stepp, M. A. (2020). Axonal debris accumulates in corneal epithelial cells after intraepithelial corneal nerves are damaged: A focused Ion Beam Scanning Electron Microscopy (FIB-SEM) study. Experimental Eye Research, 194, 107998. https://doi.org/10.1016/j.exer.2020.107998
Parlanti P, et al. Axonal Debris Accumulates in Corneal Epithelial Cells After Intraepithelial Corneal Nerves Are Damaged: a Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) Study. Exp Eye Res. 2020;194:107998. PubMed PMID: 32209319.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Axonal debris accumulates in corneal epithelial cells after intraepithelial corneal nerves are damaged: A focused Ion Beam Scanning Electron Microscopy (FIB-SEM) study. AU - Parlanti,Paola, AU - Pal-Ghosh,Sonali, AU - Williams,Alexa, AU - Tadvalkar,Gauri, AU - Popratiloff,Anastas, AU - Stepp,Mary Ann, Y1 - 2020/03/21/ PY - 2019/11/27/received PY - 2020/02/13/revised PY - 2020/03/07/accepted PY - 2020/3/27/pubmed PY - 2020/12/16/medline PY - 2020/3/27/entrez KW - Cornea KW - Corneal nerves KW - FIB-SEM KW - Mouse KW - Trephine injury SP - 107998 EP - 107998 JF - Experimental eye research JO - Exp Eye Res VL - 194 N2 - The intraepithelial corneal nerves (ICNs) that innervate the corneal epithelium are maintained through interactions with corneal epithelial cells and the extracellular matrix they produce. One to several axons bundle together within the basal cell layer and extend parallel to the ocular surface or branch and extend apically. Here we use 3-dimentional (3D) ultrastructural reconstructions of control and trephine injured mouse corneal epithelium and stroma produced using Focused Ion Beam Scanning Electron Microscope (FIB-SEM) to determine whether corneal epithelial or immune cells resident in the epithelium remove axonal debris and degrade it in their lysosomes after trephine injury to the cornea. We demonstrate that axonal fragments are internalized in the corneal epithelium and accumulate within electron dense structures consistent with lysosomes 3 h after trephine injury in both epithelial and immune cells located among the basal cells of the trephine injured cornea. Confocal imaging showed fewer CD45[+] immune cells within the corneal epithelium after trephine injury compared to controls. The resolution obtained using FIB-SEM also allowed us to show that the presence of sensory axons at the basal aspect of the epithelial basal cells close to the anterior aspect of the epithelial basement membrane (EBM) is associated with a focal reduction in EBM thickness. In addition, we show using FIB-SEM and confocal imaging that superficial trephine injuries that do not penetrate the stroma, damage the integrity of anterior stromal nerves. These studies are the first to look at the mouse cornea following nerve injury using FIB-SEM. SN - 1096-0007 UR - https://www.unboundmedicine.com/medline/citation/32209319/Axonal_debris_accumulates_in_corneal_epithelial_cells_after_intraepithelial_corneal_nerves_are_damaged:_A_focused_Ion_Beam_Scanning_Electron_Microscopy__FIB_SEM__study_ DB - PRIME DP - Unbound Medicine ER -