Expression of phosphatidyhnositol 3-kinase during EGF-stimulated wound repair in rabbit corneal epithelium

Yi Zhang, Gregory I Liou, Adarsh K Gulati, Rashid A. Akhtar

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Purpose. To investigate the effect of epidermal growth factor (EGF) on the induction of phosphatidylinositol 3-kinase (PI 3-kinase) gene expression during rabbit corneal epithelial wound repair. Methods. Epithelial wounds (6 mm in size) were created in rabbit corneas and EGF (2 μg) applied every 8 hours to one eye, and the other eye served as a control. The wound repair was monitored by staining the tissue with fluorescein followed by photography. The wound area was quantified with a computer program. At different time intervals, the rabbits were killed and the corneal epithelium used for estimation of PI 3- kinase activity, western blot analysis, or reverse transcription-polymerase chain reaction (RT-PCR). For in situ hybridization, the whole corneas were sectioned and the sections processed with PI 3-kinase mRNA probes. Results. In the untreated eye, the epithelial wound progressively healed in a time-dependent manner, with 75% of the wound closed at 48 hours post wounding. Application of EGF to the corneal epithelium further stimulated wound repair at all time intervals, and the wound was completely closed at 48 hours. Analysis of PI 3-kinase showed a time- dependent increase in its enzyme activity that was maximally increased at 36 hours, the time when the wound was nearly closed. Western blot analysis revealed increased amounts of PI 3- kinase protein during the course of wound repair. Analysis of RT-PCR products from epithelial tissues, taken at different times during wound repair, showed increased PI 3-kinase expression that was maximum at 48 hours post wounding. A visible increase in PI 3-kinase gene expression was also detected by in situ hybridization during the course of the wound repair. This expression was increased maximally by EGF at 48 hours post wounding. Conclusions. The results indicate a temporal correlation between increased activation and expression of PI 3- kinase and the epithelial wound repair. Topical application of EGF further stimulates the activity and expression of PI 3- kinase. It is suggested that PI 3- kinase and its products may play a role in EGF-induced cell proliferation during corneal epithelial wound repair.

Original languageEnglish (US)
Pages (from-to)2819-2826
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume40
Issue number12
StatePublished - Nov 1 1999

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Corneal Epithelium
Epidermal Growth Factor
Phosphotransferases
Rabbits
Wounds and Injuries
Phosphatidylinositol 3-Kinase
Phosphatidylinositol 3-Kinases
Cornea
Reverse Transcription
In Situ Hybridization
Western Blotting
Gene Expression
Polymerase Chain Reaction
Photography
Fluorescein

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Expression of phosphatidyhnositol 3-kinase during EGF-stimulated wound repair in rabbit corneal epithelium. / Zhang, Yi; Liou, Gregory I; Gulati, Adarsh K; Akhtar, Rashid A.

In: Investigative Ophthalmology and Visual Science, Vol. 40, No. 12, 01.11.1999, p. 2819-2826.

Research output: Contribution to journalArticle

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abstract = "Purpose. To investigate the effect of epidermal growth factor (EGF) on the induction of phosphatidylinositol 3-kinase (PI 3-kinase) gene expression during rabbit corneal epithelial wound repair. Methods. Epithelial wounds (6 mm in size) were created in rabbit corneas and EGF (2 μg) applied every 8 hours to one eye, and the other eye served as a control. The wound repair was monitored by staining the tissue with fluorescein followed by photography. The wound area was quantified with a computer program. At different time intervals, the rabbits were killed and the corneal epithelium used for estimation of PI 3- kinase activity, western blot analysis, or reverse transcription-polymerase chain reaction (RT-PCR). For in situ hybridization, the whole corneas were sectioned and the sections processed with PI 3-kinase mRNA probes. Results. In the untreated eye, the epithelial wound progressively healed in a time-dependent manner, with 75{\%} of the wound closed at 48 hours post wounding. Application of EGF to the corneal epithelium further stimulated wound repair at all time intervals, and the wound was completely closed at 48 hours. Analysis of PI 3-kinase showed a time- dependent increase in its enzyme activity that was maximally increased at 36 hours, the time when the wound was nearly closed. Western blot analysis revealed increased amounts of PI 3- kinase protein during the course of wound repair. Analysis of RT-PCR products from epithelial tissues, taken at different times during wound repair, showed increased PI 3-kinase expression that was maximum at 48 hours post wounding. A visible increase in PI 3-kinase gene expression was also detected by in situ hybridization during the course of the wound repair. This expression was increased maximally by EGF at 48 hours post wounding. Conclusions. The results indicate a temporal correlation between increased activation and expression of PI 3- kinase and the epithelial wound repair. Topical application of EGF further stimulates the activity and expression of PI 3- kinase. It is suggested that PI 3- kinase and its products may play a role in EGF-induced cell proliferation during corneal epithelial wound repair.",
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N2 - Purpose. To investigate the effect of epidermal growth factor (EGF) on the induction of phosphatidylinositol 3-kinase (PI 3-kinase) gene expression during rabbit corneal epithelial wound repair. Methods. Epithelial wounds (6 mm in size) were created in rabbit corneas and EGF (2 μg) applied every 8 hours to one eye, and the other eye served as a control. The wound repair was monitored by staining the tissue with fluorescein followed by photography. The wound area was quantified with a computer program. At different time intervals, the rabbits were killed and the corneal epithelium used for estimation of PI 3- kinase activity, western blot analysis, or reverse transcription-polymerase chain reaction (RT-PCR). For in situ hybridization, the whole corneas were sectioned and the sections processed with PI 3-kinase mRNA probes. Results. In the untreated eye, the epithelial wound progressively healed in a time-dependent manner, with 75% of the wound closed at 48 hours post wounding. Application of EGF to the corneal epithelium further stimulated wound repair at all time intervals, and the wound was completely closed at 48 hours. Analysis of PI 3-kinase showed a time- dependent increase in its enzyme activity that was maximally increased at 36 hours, the time when the wound was nearly closed. Western blot analysis revealed increased amounts of PI 3- kinase protein during the course of wound repair. Analysis of RT-PCR products from epithelial tissues, taken at different times during wound repair, showed increased PI 3-kinase expression that was maximum at 48 hours post wounding. A visible increase in PI 3-kinase gene expression was also detected by in situ hybridization during the course of the wound repair. This expression was increased maximally by EGF at 48 hours post wounding. Conclusions. The results indicate a temporal correlation between increased activation and expression of PI 3- kinase and the epithelial wound repair. Topical application of EGF further stimulates the activity and expression of PI 3- kinase. It is suggested that PI 3- kinase and its products may play a role in EGF-induced cell proliferation during corneal epithelial wound repair.

AB - Purpose. To investigate the effect of epidermal growth factor (EGF) on the induction of phosphatidylinositol 3-kinase (PI 3-kinase) gene expression during rabbit corneal epithelial wound repair. Methods. Epithelial wounds (6 mm in size) were created in rabbit corneas and EGF (2 μg) applied every 8 hours to one eye, and the other eye served as a control. The wound repair was monitored by staining the tissue with fluorescein followed by photography. The wound area was quantified with a computer program. At different time intervals, the rabbits were killed and the corneal epithelium used for estimation of PI 3- kinase activity, western blot analysis, or reverse transcription-polymerase chain reaction (RT-PCR). For in situ hybridization, the whole corneas were sectioned and the sections processed with PI 3-kinase mRNA probes. Results. In the untreated eye, the epithelial wound progressively healed in a time-dependent manner, with 75% of the wound closed at 48 hours post wounding. Application of EGF to the corneal epithelium further stimulated wound repair at all time intervals, and the wound was completely closed at 48 hours. Analysis of PI 3-kinase showed a time- dependent increase in its enzyme activity that was maximally increased at 36 hours, the time when the wound was nearly closed. Western blot analysis revealed increased amounts of PI 3- kinase protein during the course of wound repair. Analysis of RT-PCR products from epithelial tissues, taken at different times during wound repair, showed increased PI 3-kinase expression that was maximum at 48 hours post wounding. A visible increase in PI 3-kinase gene expression was also detected by in situ hybridization during the course of the wound repair. This expression was increased maximally by EGF at 48 hours post wounding. Conclusions. The results indicate a temporal correlation between increased activation and expression of PI 3- kinase and the epithelial wound repair. Topical application of EGF further stimulates the activity and expression of PI 3- kinase. It is suggested that PI 3- kinase and its products may play a role in EGF-induced cell proliferation during corneal epithelial wound repair.

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