Regeneration of rabbit cornea following excimer laser photorefractive keratectomy: A study on gap junctions, epithelial junctions and epidermal growth factor receptor expression in correlation with cell proliferation

I. Ratkay-Traub, B. Hopp, Zs Bor, L. Dux, D. L. Becker, T. Krenacs

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Abstract

Corneal wound repair was investigated in rabbits following excimer laser ablation of a 6 mm diameter and 90 μm deep disc. In the healing process particular attention was focused on the epithelium where gap junction expression and the rearrangement of desmosomes and hemidesmosomes were correlated with cell proliferation and epidermal growth factor receptor expression. Immunofluorescence-based confocal laser scanning microscopy, semithin resin section morphology and electron microscopy were utilized. In resting cornea two isotypes of gap junctions, confined to different regions in the same basal epithelial cells, were detected. Particulate connexin43 (α1) immunostaining was concentrated on the apical while the connexin26 type (β2) in the baso-latheral cell membranes. This is the first report of connexin26 in the cornea. Connexin43 was found also in corneal keratocytes and endothelial cell. Since the two connexins do not form functioning heteromeric channels and have selective permeabilities they may serve alternative pathways for direct cell-cell communication in the basal cell layer. During regeneration both connexins were expressed throughout the corneal epithelium including the migrating cells. They also showed transient up-regulation 24 hr after wounding in the form of overlapping relocation to the upper cell layers. At this time, basal epithelial ceils at the limbal region, adjacent to the wound and those migrating over the wounded area all expressed membrane bound epidermal growth factor receptor and they were highly proliferating. In conclusion, like in other stratified epithelia connexin26 is also expressed in the cornea. Transient up-regulation and relocation of connexins within the regenerating epithelium may reflect the involvement of direct cell-cell communication in corneal wound healing. Mitotic activity in the migrating corneal epithelial ceils is also a novel finding which is probably the sign of the excessive demand for new epithelial cells in larger wounds not met alone by the proliferating limbal stock.

Original languageEnglish
Pages (from-to)291-302
Number of pages12
JournalExperimental Eye Research
Volume73
Issue number3
DOIs
Publication statusPublished - 2001

Fingerprint

Laser Corneal Surgery
Photorefractive Keratectomy
Excimer Lasers
Gap Junctions
Epidermal Growth Factor Receptor
Cornea
Regeneration
Cell Proliferation
Connexins
Rabbits
Connexin 43
Epithelium
Cell Communication
Wounds and Injuries
Corneal Keratocytes
Up-Regulation
Epithelial Cells
Hemidesmosomes
Desmosomes
Corneal Epithelium

Keywords

  • Connexin26
  • Connexin43
  • Cornea
  • Desmosomes
  • Epidermal growth factor receptor
  • Epithelial cell proliferation
  • Gap junctions
  • Hemidesmosomes
  • Laser ablation
  • Regeneration

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

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title = "Regeneration of rabbit cornea following excimer laser photorefractive keratectomy: A study on gap junctions, epithelial junctions and epidermal growth factor receptor expression in correlation with cell proliferation",
abstract = "Corneal wound repair was investigated in rabbits following excimer laser ablation of a 6 mm diameter and 90 μm deep disc. In the healing process particular attention was focused on the epithelium where gap junction expression and the rearrangement of desmosomes and hemidesmosomes were correlated with cell proliferation and epidermal growth factor receptor expression. Immunofluorescence-based confocal laser scanning microscopy, semithin resin section morphology and electron microscopy were utilized. In resting cornea two isotypes of gap junctions, confined to different regions in the same basal epithelial cells, were detected. Particulate connexin43 (α1) immunostaining was concentrated on the apical while the connexin26 type (β2) in the baso-latheral cell membranes. This is the first report of connexin26 in the cornea. Connexin43 was found also in corneal keratocytes and endothelial cell. Since the two connexins do not form functioning heteromeric channels and have selective permeabilities they may serve alternative pathways for direct cell-cell communication in the basal cell layer. During regeneration both connexins were expressed throughout the corneal epithelium including the migrating cells. They also showed transient up-regulation 24 hr after wounding in the form of overlapping relocation to the upper cell layers. At this time, basal epithelial ceils at the limbal region, adjacent to the wound and those migrating over the wounded area all expressed membrane bound epidermal growth factor receptor and they were highly proliferating. In conclusion, like in other stratified epithelia connexin26 is also expressed in the cornea. Transient up-regulation and relocation of connexins within the regenerating epithelium may reflect the involvement of direct cell-cell communication in corneal wound healing. Mitotic activity in the migrating corneal epithelial ceils is also a novel finding which is probably the sign of the excessive demand for new epithelial cells in larger wounds not met alone by the proliferating limbal stock.",
keywords = "Connexin26, Connexin43, Cornea, Desmosomes, Epidermal growth factor receptor, Epithelial cell proliferation, Gap junctions, Hemidesmosomes, Laser ablation, Regeneration",
author = "I. Ratkay-Traub and B. Hopp and Zs Bor and L. Dux and Becker, {D. L.} and T. Krenacs",
year = "2001",
doi = "10.1006/exer.2001.1040",
language = "English",
volume = "73",
pages = "291--302",
journal = "Experimental Eye Research",
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T1 - Regeneration of rabbit cornea following excimer laser photorefractive keratectomy

T2 - A study on gap junctions, epithelial junctions and epidermal growth factor receptor expression in correlation with cell proliferation

AU - Ratkay-Traub, I.

AU - Hopp, B.

AU - Bor, Zs

AU - Dux, L.

AU - Becker, D. L.

AU - Krenacs, T.

PY - 2001

Y1 - 2001

N2 - Corneal wound repair was investigated in rabbits following excimer laser ablation of a 6 mm diameter and 90 μm deep disc. In the healing process particular attention was focused on the epithelium where gap junction expression and the rearrangement of desmosomes and hemidesmosomes were correlated with cell proliferation and epidermal growth factor receptor expression. Immunofluorescence-based confocal laser scanning microscopy, semithin resin section morphology and electron microscopy were utilized. In resting cornea two isotypes of gap junctions, confined to different regions in the same basal epithelial cells, were detected. Particulate connexin43 (α1) immunostaining was concentrated on the apical while the connexin26 type (β2) in the baso-latheral cell membranes. This is the first report of connexin26 in the cornea. Connexin43 was found also in corneal keratocytes and endothelial cell. Since the two connexins do not form functioning heteromeric channels and have selective permeabilities they may serve alternative pathways for direct cell-cell communication in the basal cell layer. During regeneration both connexins were expressed throughout the corneal epithelium including the migrating cells. They also showed transient up-regulation 24 hr after wounding in the form of overlapping relocation to the upper cell layers. At this time, basal epithelial ceils at the limbal region, adjacent to the wound and those migrating over the wounded area all expressed membrane bound epidermal growth factor receptor and they were highly proliferating. In conclusion, like in other stratified epithelia connexin26 is also expressed in the cornea. Transient up-regulation and relocation of connexins within the regenerating epithelium may reflect the involvement of direct cell-cell communication in corneal wound healing. Mitotic activity in the migrating corneal epithelial ceils is also a novel finding which is probably the sign of the excessive demand for new epithelial cells in larger wounds not met alone by the proliferating limbal stock.

AB - Corneal wound repair was investigated in rabbits following excimer laser ablation of a 6 mm diameter and 90 μm deep disc. In the healing process particular attention was focused on the epithelium where gap junction expression and the rearrangement of desmosomes and hemidesmosomes were correlated with cell proliferation and epidermal growth factor receptor expression. Immunofluorescence-based confocal laser scanning microscopy, semithin resin section morphology and electron microscopy were utilized. In resting cornea two isotypes of gap junctions, confined to different regions in the same basal epithelial cells, were detected. Particulate connexin43 (α1) immunostaining was concentrated on the apical while the connexin26 type (β2) in the baso-latheral cell membranes. This is the first report of connexin26 in the cornea. Connexin43 was found also in corneal keratocytes and endothelial cell. Since the two connexins do not form functioning heteromeric channels and have selective permeabilities they may serve alternative pathways for direct cell-cell communication in the basal cell layer. During regeneration both connexins were expressed throughout the corneal epithelium including the migrating cells. They also showed transient up-regulation 24 hr after wounding in the form of overlapping relocation to the upper cell layers. At this time, basal epithelial ceils at the limbal region, adjacent to the wound and those migrating over the wounded area all expressed membrane bound epidermal growth factor receptor and they were highly proliferating. In conclusion, like in other stratified epithelia connexin26 is also expressed in the cornea. Transient up-regulation and relocation of connexins within the regenerating epithelium may reflect the involvement of direct cell-cell communication in corneal wound healing. Mitotic activity in the migrating corneal epithelial ceils is also a novel finding which is probably the sign of the excessive demand for new epithelial cells in larger wounds not met alone by the proliferating limbal stock.

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KW - Epidermal growth factor receptor

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KW - Hemidesmosomes

KW - Laser ablation

KW - Regeneration

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