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Cellular distribution of lens epithelium-derived growth factor (LEDGF) in the rat eye: loss of LEDGF from nuclei of differentiating cells

Histochemistry and Cell Biology volume 119, pages 289–299 (2003)Cite this article

Abstract

Lens epithelium-derived growth factor (LEDGF) enhances the survival and growth of cells. To understand LEDGF's spatial localization and its putative function(s) during proliferation and differentiation, we localized LEDGF during terminal differentiation in whole rat lenses, lens epithelial cell (LEC) explants stimulated with FGF-2, and insulin, iris, human LECs with lentoids. In addition, intracellular localization of LEDGF was performed in other ocular tissues: ciliary body, retina, and cornea. We found the immunopositivity of nuclear LEDGF decreased in LECs of the equatorial region. In contrast, immunopositivity of LEDGF was detected in the cytoplasm of LECs and superficial fiber cells. After treating LEC explants with FGF-2 and insulin, which are known to be differentiating factors for LECs, the nuclei of these cells showed no LEDGF immunopositivity, but explants did express p57kip2, a differentiation marker protein. Also, immunopositive LEDGF was not detected in the nuclei of differentiated cells, lentoid body, and corneal epithelial cells. This demonstrated that the loss of LEDGF from the nucleus may be associated with the process of terminal differentiation that might be in some way common with the biochemical mechanisms of apoptosis. The spatial and temporal distribution of LEDGF in the present study also provides a vision for further investigation as to how this protein is involved in cell fate determination.

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References

  • Arita T, Lin LR, Reddy VN (1988) Differentiation of human lens epithelial cells in tissue culture. Exp Eye Res 47:905–910

    CAS  PubMed  Google Scholar 

  • Arita T, Lin LR, Susan SR, Reddy VN (1990) Enhancement of differentiation of human lens epithelium in tissue culture by changes in cell-substrate adhesion. Invest Ophthalmol Vis Sci 31:2395–2404

    CAS  PubMed  Google Scholar 

  • Balaram M, Tung WH, Kuszak J, Ayaki M, Shinohara T, Chylack LT Jr (2000) Non-contact specular microscopy of human lens epithelium. Invest Ophthalmol Vis Sci 41:474–481

    CAS  PubMed  Google Scholar 

  • Bassnett S, Beebe DC (1992) Coincident loss of mitochondria and nuclei during lens fiber cell differentiation. Dev Dyn 194:85–93

    CAS  PubMed  Google Scholar 

  • Beere HM, Wolf BB, Cain K, Mosser DD, Mahbboudi A, Kuwana, T, Railor P, Moromoto RI, Cohen GM, Green DR (2000) Heat shock protein 70 inhibits apoptosis by preventing the recruitment of procaspase-9 to the Apaf-1 apoptosome. Nat Cell Biol 2:469–475

    Google Scholar 

  • Bradford MM (1976) A rapid sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Bruey JM, Ducasse C, Bonniaud P, Ravagnan L, Susin SA, Siaz-Latoud C, Gurbuxani S, Arrigo AP, Kroemer G, Solary E, Garrido C (2000) Hsp-27 negatively regulates cell death by interacting with cytochrome c. Nat Cell Biol 2:645–652

    Google Scholar 

  • Carey RG, Li B, DiCicco-Bloom E (2002) Pituitary adenylate cyclase activating polypeptide anti-mitogenic signaling in cerebral cortical progenitors is regulated by p57Kip2-dependent CDK2 activity. J Neurosci 22:1583–1591

    CAS  PubMed  Google Scholar 

  • Chamberlain CG, McAvoy JW (1987) Evidence that fibroblast growth factor promotes lens fiber differentiation. Curr Eye Res 6:1165–1168

    CAS  PubMed  Google Scholar 

  • Chamberlain CG, McAvoy JW (1989) Induction of lens fiber differentiation by acidic and basic fibroblast growth factor (FGF). Growth Factors 1:125–134

    CAS  PubMed  Google Scholar 

  • Chamberlain CG, McAvoy JW, Richardson NA (1991) The effects of insulin and basic fibroblast growth factor on fiber differentiation in rat lens epithelial explants. Growth Factors 4:183–188

    CAS  PubMed  Google Scholar 

  • Dahm R (1999) Lens fiber cell differentiation: a link with apoptosis. Ophthalmic Res 31:163–183

    Article  CAS  PubMed  Google Scholar 

  • Dahm R, Gribbon C, Quinlan RA, Prescott AR (1998) Changes in nucleolar and coiled body compartments precede lamina and chromatin reorganization during fiber cell denucleation in the bovine lens. Eur J Cell Biol 75:237–246

    CAS  PubMed  Google Scholar 

  • Fatma N, Singh DP, Shinohara T, Chylack LT Jr (2000) Heparin's roles in stabilizing, potentiating and transporting LEDGF into the nucleus. Invest Ophthalmol Vis Sci 41:2648–2657

    CAS  PubMed  Google Scholar 

  • Fatma N, Singh DP, Shinohara T, Chylack LT Jr (2001) Transcriptional regulation of the antioxidant protein 2 gene, a thiol-specific antioxidant, by lens epithelium-derived growth factor to protect cells from oxidative stress. J Biol Chem 276:48899–48907

    Article  CAS  PubMed  Google Scholar 

  • Ge H, Si Y, Roeder RG (1998a) Isolation of cDNAs encoding novel transcription co-activators p52 and p75 reveals an alternate regulatory mechanism of transcriptional activation. EMBO J 17:6723–6729

    Article  CAS  PubMed  Google Scholar 

  • Ge H, Si Y, Wolffe AP (1998b) A novel transcriptional coactivator, p52, functionally interacts with the essential splicing factor ASF/SF2. Mol Cell 2:751–759

    CAS  PubMed  Google Scholar 

  • Hatada I, Mukai T (1995) Genomic imprinting of p57kip2, a cyclin-dependent kinase inhibitor, in mouse. Nat Genet 11:204–206

    CAS  PubMed  Google Scholar 

  • Hengst L, Duvic V, Slingerland JM, Lees E, Red SI (1994) A cell cycle-regulated inhibitor of cyclin-dependent kinases. Proc Natl Acad Sci U S A 91:5291–5295

    CAS  PubMed  Google Scholar 

  • Hettmann T, Barton K, Leiden JM (2000) Microphthalmia due to p53-mediated apoptosis of anterior lens epithelial cells in mice lacking the CREB-2 transcription factor. Dev Biol 222:110–123

    Article  CAS  PubMed  Google Scholar 

  • Ibaraki N, Lin LR, Reddy VN (1995) Effects of growth factors on proliferation and differentiation in human lens epithelial cells in early subculture. Invest Ophthalmol Vis Sci 36:2304–2312

    CAS  PubMed  Google Scholar 

  • Ishizaki Y, Voyvodic JT, BurneJF, Raff MC (1993) Control of lens epithelial cell survival. J Cell Biol 121:899–908

    CAS  PubMed  Google Scholar 

  • Ishizaki Y, Jacobson MD, Raff MC (1998) A role of caspases in lens fiber differentiation. J Cell Biol 140:153–158

    Article  CAS  PubMed  Google Scholar 

  • Iuchi S, Green H (1997) Nuclear localization of basonuclin in human keratinocytes and the role of phosphorylation. Proc Natl Acad Sci U S A 94:7948–7953

    Article  CAS  PubMed  Google Scholar 

  • Klok E, Lubsen NH, Chamberlain CG, McAvoy JW (1998) Induction and maintenance of differentiation of rat lens epithelium by FGF2, insulin and IGF-1. Exp Eye Res 67:425–431

    Article  CAS  PubMed  Google Scholar 

  • Kubo E, Fatma N, Shinohara T, Chylack LT Jr, Akagi Y, Singh DP (2002) Transactivation of involucrin, a marker of differentiation in keratinocytes, by lens epithelium derived growth factor (LEDGF). J Mol Biol 320:1053–1063

    Article  CAS  PubMed  Google Scholar 

  • Lee MH, Reynisdottir I, Massague J (1995) Cloning of p57kip2, a cyclin-dependent kinase inhibitor with unique domain structure and tissue distribution. Genes Dev 9:639–649

    PubMed  Google Scholar 

  • Li DW, Xiang H, Mao YW, Wang J, Fass U, Zhang XY, Xu C (2001) Caspase-3 is actively involved in okadaic acid-induced lens epithelial cell apoptosis. Exp Cell Res 266:279–291

    Article  CAS  PubMed  Google Scholar 

  • Lovicu FJ, McAvoy JW (1989) Structural analysis of lens epithelial explants induced to differentiate into fibers by fibroblast growth factor (FGF). Exp Eye Res 49:479–494

    CAS  PubMed  Google Scholar 

  • Lovicu FJ, McAvoy JW (1999) Spatial and temporal expression of p57kip2 during murine lens development Mech Dev 86:165–169

    Article  CAS  Google Scholar 

  • Machida S, Chaudhary P, Shinohara T, Singh DP, Reddy VN, Chylack LT Jr, Sieving PA, Bush RA (2001) Lens epithelium-derived growth factor promotes photoreceptor survival in light-damaged and RCS rats. Invest Ophthalmol Vis Sci 42:1087–1095

    CAS  PubMed  Google Scholar 

  • Matsui H, Lin LR, Singh DP, Shinohara T, Reddy VN (2001) Lens epithelium-derived growth factor: increased survival and decreased DNA breakage of human RPE cells induced by oxidative stress. Invest Ophthalmol Vis Sci 42:2935–2941

    CAS  PubMed  Google Scholar 

  • Matsuoka S, Edwards MC, Bai C, Parker S, Zhang P, Baldini A, Harper JW, Elledge SJ (1995) p57kip2, a structurally distinct member of the p21kip1 Cdk inhibitor family, is a candidate tumor suppressor gene. Genes Dev 9:650–662

    CAS  PubMed  Google Scholar 

  • McAvoy JW, Chamberlain CG (1989) Fibroblast growth factor (FGF) induced different responses in lens epithelial cells depending on its concentration. Development 107:221–228

    CAS  PubMed  Google Scholar 

  • Nakamura M, Singh DP, Kubo E, Chylack LT Jr, Shinohara T (2000) LEDGF: survival of embryonic chick retinal photoreceptor cells. Invest Ophthalmol Vis Sci 41:1168–1175

    CAS  PubMed  Google Scholar 

  • Piatigorsky J (1981) Lens differentiation in vertebrates: a review of cellular and molecular features. Differentiation 19:134–153

    CAS  PubMed  Google Scholar 

  • Raff MC (1992) Social controls on cell survival and cell death. Nature 356:397–400

    Google Scholar 

  • Rampalli AM, Zelenka PS (1995) Insulin regulates expression of c-fos and c-jun and suppresses apoptosis of lens epithelial cells. Cell Growth Differ 6:945–953

    CAS  PubMed  Google Scholar 

  • Renaud F, Oliver L, Desset, Tassin J, Romquin N, Courtois Y, Laurent M (1994) Up-regulation of FGF-1 expression in quiescent cells is related to cell survival. J Cell Physiol 158:435–443

    CAS  PubMed  Google Scholar 

  • Saleh A, Srinivasula SM, Balki L, Robbins PD, Alnemri ES (2000) Negative regulation of Apaf-1 apoptosome by Hsp70. Nat Cell Biol 2:476–483

    Google Scholar 

  • Sharma P, Singh DP, Fatma N, Chylack LT Jr, Shinohara T (2000) Activation of LEDGF gene by thermal- and oxidative-stresses. Biochem Biophys Res Commun 276:1320–1324

    Article  CAS  PubMed  Google Scholar 

  • Singh DP, Ohguro N, Chylack LT Jr, Shinohara T (1999) Lens epithelium-derived growth factor: increased resistance to thermal and oxidative stresses. Invest Ophthalmol Vis Sci 40:1444–1451

    CAS  PubMed  Google Scholar 

  • Singh DP, Ohguro N, Kikuchi T, Sueno T, Reddy VN, Yuge K, Ko V, Chylack LT Jr, Shinohara T (2000a) Lens epithelium-derived growth factor: effects of growth and survival of lens epithelial cells, keratinocytes, and fibroblasts. Biochem Biophys Res Commun 267:373–381

    Article  CAS  PubMed  Google Scholar 

  • Singh DP, Kimura A, Chylack LT Jr, Shinohara T (2000b) Lens epithelium-derived growth factor (LEDGF/p75) and p52 are derived from a single gene by alternative splicing. Gene 242:265–273

    Article  CAS  PubMed  Google Scholar 

  • Singh DP, Fatma N, Kimura A, Chylack LT Jr, Shinohara T (2001a) LEDGF binds to heat shock and stress-related element to activate the expression of stress-related genes. Biochem Biophys Res Commun 283:943–955

    Article  CAS  PubMed  Google Scholar 

  • Singh DP, Fatma N, Chylack LT Jr, Shinohara T (2001b) Distinct nuclear localization signal and DNA-binding domains of LEDGF. Invest Ophthalmol Vis Sci (suppl) 42:2909

    Google Scholar 

  • Stolen CM, Jackson MW, Griep AE (1997) Overexpression of FGF-2 modulates fiber cell differentiation and survival in the mouse lens. Development 124:4009–4017

    CAS  PubMed  Google Scholar 

  • Tan X, Martin SJ, Green DR, Wang JY (1997) Degradation of retinoblastoma protein in tumor necrosis factor- and CD95-induced cell death. J Biol Chem 212:9613–9616

    Google Scholar 

  • Tseng H, Matsuzaki K, Lavker RM (1999) Basonuclin in murine corneal and lens epithelia correlates with cellular maturation and proliferative ability. Differentiation 65:221–227

    Article  CAS  PubMed  Google Scholar 

  • Wride MA (2000) Apoptosis as seen through the lens. Apoptosis 5:203–209

    Article  CAS  PubMed  Google Scholar 

  • Wride MA, Sanders EJ (1998) Nuclear degeneration in the developing lens and its regulation by TNF alpha. Exp Eye Res 66:371–383

    Article  CAS  PubMed  Google Scholar 

  • Wu X, Daniels T, Molinaro C, Lilly MB, Casiano CA (2002) Caspase cleavage of the nuclear autoantigens LEDGF/p75 abrogates its pro-survival function: implication for autoimmunity in atopic disorders. Cell Death Differ 9:915–925

    Article  CAS  PubMed  Google Scholar 

  • Zohang P, Liégeois NJ, Wong C, Finegold M, Hon H, Thompson JC, Silverman A, Harper JW, DePinho RA, Elledge SJ (1997) Altered cell differentiation and proliferation in mice lacking p57kip2 indicates a role in Beckwith-Wiedermann syndrome. Nature 387:151–158

    Google Scholar 

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Acknowledgements

The work was supported by NIH grants EY12015, EY10824, EY13394, and EY07003, and the Massachusetts Lions Eye Research Fund.

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Affiliations

  1. Department of Ophthalmology, Room 2009, University of Nebraska Medical Center, Omaha, NE 68198-6395, USA

    Dhirendra P. Singh, Nigar Fatma & Toshimichi Shinohara

  2. The Center for Ophthalmic Research, Brigham and Women's Hospital, Havard Medical School, 221 Longwood Avenue, Boston, MA 02115, USA

    Eri Kubo, Dhirendra P. Singh, Nigar Fatma, Toshimichi Shinohara & Leo T. Chylack Jr

  3. Department of Ophthalmology, Fukui Medical University, Fukui, Japan

    Eri Kubo

  4. Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA

    Eri Kubo & Peggy Zelenka

  5. Kellogg Eye Center, Department of Ophthalmology and Visual Science, University of Michigan, Ann Arbor, Michigan, USA

    Venkat N. Reddy

  6. Wittson Hall, Room 2009, University of Nebraska Medical Center

    Dhirendra P. Singh

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  1. Eri Kubo
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Correspondence to Dhirendra P. Singh.

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Kubo, E., Singh, D.P., Fatma, N. et al. Cellular distribution of lens epithelium-derived growth factor (LEDGF) in the rat eye: loss of LEDGF from nuclei of differentiating cells. Histochem Cell Biol 119, 289–299 (2003). https://doi.org/10.1007/s00418-003-0518-3

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  • DOI: https://doi.org/10.1007/s00418-003-0518-3

Keywords

  • LEDGF
  • Immunohistochemistry
  • Differentiation
  • In situ hybridization
  • Lentoid body
  • LEC explants