Phosphorylation of p27(KIP1) in the mitotic cells of the corneal epithelium

Satoru Kase, Kazuhiko Yoshida, Kazuhiro Ohgami, Kenji Shiratori, Shigeaki Ohno, Keiichi I. Nakayama

Research output: Contribution to journalArticle

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Abstract

Purpose: The mechanism in regulation of the cell cycle and proliferation of corneal epithelium in the homeostatic ocular surface remains unclear. The aim of this study is to examine the expression of p27(KIP1) and its phosphorylation in corneal epithelium. Methods: The eyes of C57BL/6 mice (7 weeks old) were enucleated. Formalin-fixed and paraffin-embedded tissue sections were examined using immunohistochemistry with anti-p27(KIP1), threonine 187 phosphorylated p27(KIP1) (T187-phospho-p27), and phosphorylated Histon H3 (pHiston H3) antibodies. Anti-T187-phospho-p27 and anti-pHiston H3 polyclonal antibodies were used for parallel immunofluorescent staining. Results: pHiston H3-immunopositive cells were noted in basal cells of the corneal epithelium. At high magnification of DAPI nuclear staining, mitotic and non-mitotic cells were observed in corneal basal layer. p27(KIP1)-positive nuclei were detected in corneal basal cells, where non-mitotic basal cells were located. In contrast, mitotic cells showed under detectable level on p27(KIP1) immunoreactivity. Immunoreactivity for T187-phospho-p27 was detected in basal cells of the corneal epithelium. At high magnification, it was confirmed that the immunopositive cells were mitotic cells. Immunoreactivity of T187-phospho-p27 as well as pHiston H3 was localized in the same corneal basal cells using double-staining immunohistochemistry. Conclusions: These results suggested that degradation of p27(KIP1) regulates progression into mitosis in corneal basal cells.

Original languageEnglish
Pages (from-to)307-312
Number of pages6
JournalCurrent Eye Research
Volume31
Issue number4
DOIs
Publication statusPublished - May 1 2006

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Corneal Epithelium
Phosphorylation
Staining and Labeling
Immunohistochemistry
Antibodies
Threonine
Inbred C57BL Mouse
Mitosis
Paraffin
Formaldehyde
Cell Cycle
Cell Proliferation

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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Phosphorylation of p27(KIP1) in the mitotic cells of the corneal epithelium. / Kase, Satoru; Yoshida, Kazuhiko; Ohgami, Kazuhiro; Shiratori, Kenji; Ohno, Shigeaki; Nakayama, Keiichi I.

In: Current Eye Research, Vol. 31, No. 4, 01.05.2006, p. 307-312.

Research output: Contribution to journalArticle

Kase, Satoru ; Yoshida, Kazuhiko ; Ohgami, Kazuhiro ; Shiratori, Kenji ; Ohno, Shigeaki ; Nakayama, Keiichi I. / Phosphorylation of p27(KIP1) in the mitotic cells of the corneal epithelium. In: Current Eye Research. 2006 ; Vol. 31, No. 4. pp. 307-312.
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AB - Purpose: The mechanism in regulation of the cell cycle and proliferation of corneal epithelium in the homeostatic ocular surface remains unclear. The aim of this study is to examine the expression of p27(KIP1) and its phosphorylation in corneal epithelium. Methods: The eyes of C57BL/6 mice (7 weeks old) were enucleated. Formalin-fixed and paraffin-embedded tissue sections were examined using immunohistochemistry with anti-p27(KIP1), threonine 187 phosphorylated p27(KIP1) (T187-phospho-p27), and phosphorylated Histon H3 (pHiston H3) antibodies. Anti-T187-phospho-p27 and anti-pHiston H3 polyclonal antibodies were used for parallel immunofluorescent staining. Results: pHiston H3-immunopositive cells were noted in basal cells of the corneal epithelium. At high magnification of DAPI nuclear staining, mitotic and non-mitotic cells were observed in corneal basal layer. p27(KIP1)-positive nuclei were detected in corneal basal cells, where non-mitotic basal cells were located. In contrast, mitotic cells showed under detectable level on p27(KIP1) immunoreactivity. Immunoreactivity for T187-phospho-p27 was detected in basal cells of the corneal epithelium. At high magnification, it was confirmed that the immunopositive cells were mitotic cells. Immunoreactivity of T187-phospho-p27 as well as pHiston H3 was localized in the same corneal basal cells using double-staining immunohistochemistry. Conclusions: These results suggested that degradation of p27(KIP1) regulates progression into mitosis in corneal basal cells.

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