Expression and regulation of cornified envelope proteins in human corneal epithelium.

PURPOSE Stratified squamous epithelial cells assemble a specialized protective barrier structure on their periphery, termed the cornified envelope. The purpose of this study was to evaluate the presence and distribution of cornified envelope precursors in human corneal epithelium, their expression in human corneal epithelial cell cultures, and the effect of ultraviolet radiation (UVB) and transglutaminase (TG) inhibition on their expression. METHODS Tissue distribution of small proline-rich proteins (SPRRs) and filaggrin and involucrin was studied in human cornea sections by immunofluorescence staining. Primary human corneal epithelial cells (HCECs) from limbal explants were used in cell culture experiments. A single dose of UVB at 20 mJ/cm2 was used to stimulate these cells, in the presence or absence of mono-dansyl cadaverine (MDC), a TG inhibitor. SPRR2 and involucrin protein levels were studied by immunofluorescence staining and Western blot analysis. Gene expression of 12 proteins was investigated by semiquantitative reverse transcription-polymerase chain reaction. RESULTS In human cornea tissue, SPRR1, SPRR2, filaggrin, and involucrin protein expression were detected in the central and peripheral corneal and limbal epithelium. In HCECs, SPRR2 and involucrin proteins were detected in the cytosolic fraction, and involucrin levels increased after UVB. Both SPRR2 and involucrin levels accumulated in the presence of MDC. Nine genes including involucrin, SPRR (types 1A, 1B, 2A, 2B, and 3), late envelope protein (LEP) 1 and 16, and filaggrin were expressed by HCECs. SPRR 4, loricrin, and LEP 6 transcripts were not detected. UVB downregulated SPRR (2A, 2B) and LEP 1 transcripts. CONCLUSIONS Various envelope precursors are expressed in human corneal epithelium and in HCECs, acute UVB stress differentially alters their expression in HCECs. The expression of envelope precursors and their rapid modulation by UVB supports the role of these proteins in the regulation of ocular surface stress. TG function may be relevant in the regulation of soluble precursors in UVB-stimulated corneal epithelium.

[1]  Seppo Ylä-Herttuala,et al.  Analysis of gene and protein expression during monocyte-macrophage differentiation and cholesterol loading--cDNA and protein array study. , 2005, Atherosclerosis.

[2]  R. Eckert,et al.  An involucrin promoter AP1 transcription factor binding site is required for expression of involucrin in the corneal epithelium in vivo. , 2005, Investigative ophthalmology & visual science.

[3]  De-Quan Li,et al.  ABCG2 Transporter Identifies a Population of Clonogenic Human Limbal Epithelial Cells , 2005, Stem cells.

[4]  Y. Ohashi,et al.  Expression of Keratinocyte Transglutaminase in Cornea of Vitamin A–Deficient Rats , 2005, Current eye research.

[5]  De-Quan Li,et al.  Experimental dry eye stimulates production of inflammatory cytokines and MMP-9 and activates MAPK signaling pathways on the ocular surface. , 2004, Investigative ophthalmology & visual science.

[6]  J. Ross,et al.  Small proline‐rich protein 1A is a gp130 pathway‐ and stress‐inducible cardioprotective protein , 2004, The EMBO journal.

[7]  S. Tseng,et al.  Calcium-induced abnormal epidermal-like differentiation in cultures of mouse corneal-limbal epithelial cells. , 2004, Investigative ophthalmology & visual science.

[8]  U. Müller-Ladner,et al.  Discrepancy between mRNA and protein expression of tumour suppressor maspin in synovial tissue may contribute to synovial hyperplasia in rheumatoid arthritis , 2004, Annals of the rheumatic diseases.

[9]  De-Quan Li,et al.  Phenotypic characterization of human corneal epithelial cells expanded ex vivo from limbal explant and single cell cultures. , 2004, Experimental eye research.

[10]  De-Quan Li,et al.  Characterization of Putative Stem Cell Phenotype in Human Limbal Epithelia , 2004, Stem cells.

[11]  F. Bernerd,et al.  Ultraviolet B induces hyperproliferation and modification of epidermal differentiation in normal human skin grafted on to nude mice , 2004, The British journal of dermatology.

[12]  R. Eckert,et al.  Regulation of involucrin expression in normal human corneal epithelial cells: a role for activator protein one. , 2004, Investigative ophthalmology & visual science.

[13]  K. Tsubota,et al.  UV dose-dependent caspase activation in a corneal epithelial cell line , 2004, Current eye research.

[14]  J. Segre.  Complex redundancy to build a simple epidermal permeability barrier. , 2003, Current opinion in cell biology.

[15]  Luo Lu,et al.  UV-induced signaling pathways associated with corneal epithelial cell apoptosis. , 2003, Investigative ophthalmology & visual science.

[16]  S. Tseng,et al.  Stromal niche controls the plasticity of limbal and corneal epithelial differentiation in a rabbit model of recombined tissue. , 2003, Investigative ophthalmology & visual science.

[17]  R. Eckert,et al.  A Regulatory Role for p38δ MAPK in Keratinocyte Differentiation , 2003, Journal of Biological Chemistry.

[18]  S. Falkow,et al.  Distinct gene expression profiles characterize the histopathological stages of disease in Helicobacter-induced mucosa-associated lymphoid tissue lymphoma , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[19]  F. Chaves,et al.  Levels of mucin gene expression in normal human conjunctival epithelium in vivo , 2003, Current eye research.

[20]  C. Byrne,et al.  Differentially expressed late constituents of the epidermal cornified envelope , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[21]  C. Backendorf,et al.  SPRR4, a novel cornified envelope precursor: UV-dependent epidermal expression and selective incorporation into fragile envelopes. , 2001, Journal of cell science.

[22]  D. Nižetić,et al.  Structural Organization and Regulation of the Small Proline-rich Family of Cornified Envelope Precursors Suggest a Role in Adaptive Barrier Function* , 2001, The Journal of Biological Chemistry.

[23]  A. Dota,et al.  Elevated expression of transglutaminase 1 and keratinization-related proteins in conjunctiva in severe ocular surface disease. , 2001, Investigative ophthalmology & visual science.

[24]  I. Marenholz,et al.  Identification of human epidermal differentiation complex (EDC)-encoded genes by subtractive hybridization of entire YACs to a gridded keratinocyte cDNA library. , 2001, Genome research.

[25]  J. Gordon,et al.  Molecular analysis of commensal host-microbial relationships in the intestine. , 2001, Science.

[26]  R. Pfundt,et al.  In situ demonstration of phosphorylated c‐jun and p38 MAP kinase in epidermal keratinocytes following ultraviolet B irradiation of human skin , 2001, The Journal of pathology.

[27]  S. Shen,et al.  Facilitated wound healing by activation of the Transglutaminase 1 gene. , 2000, The American journal of pathology.

[28]  H. Green,et al.  Targeted Ablation of the Murine Involucrin Gene , 2000, The Journal of cell biology.

[29]  D. Hohl,et al.  Expression of small proline rich proteins in neoplastic and inflammatory skin diseases , 2000, The British journal of dermatology.

[30]  B. Sola,et al.  Transcriptional and post‐transcriptional mechanisms induce cyclin‐D1 over‐expression in B‐chronic lymphoproliferative disorders , 1999, International journal of cancer.

[31]  W. Idler,et al.  Structural and Transglutaminase Substrate Properties of the Small Proline-rich 2 Family of Cornified Cell Envelope Proteins* , 1998, The Journal of Biological Chemistry.

[32]  P. Steinert,et al.  Biochemical Evidence That Small Proline-rich Proteins and Trichohyalin Function in Epithelia by Modulation of the Biomechanical Properties of Their Cornified Cell Envelopes* , 1998, The Journal of Biological Chemistry.

[33]  S. Grether-Beck,et al.  Ultraviolet A radiation-induced expression of human genes: molecular and photobiological mechanisms. , 1997, Biological chemistry.

[34]  R. Eckert,et al.  S100A11, S100A10, Annexin I, Desmosomal Proteins, Small Proline-rich Proteins, Plasminogen Activator Inhibitor-2, and Involucrin Are Components of the Cornified Envelope of Cultured Human Epidermal Keratinocytes* , 1997, The Journal of Biological Chemistry.

[35]  N. Hoogerbrugge,et al.  Growth hormone restores hepatic lipase mRNA levels but the translation is impaired in hepatocytes of hypothyroid rats. , 1997, Biochimica et biophysica acta.

[36]  F. Bernerd,et al.  Successive alteration and recovery of epidermal differentiation and morphogenesis after specific UVB-damages in skin reconstructed in vitro. , 1997, Developmental biology.

[37]  F. Castro‐Muñozledo,et al.  Development of a spontaneous permanent cell line of rabbit corneal epithelial cells that undergoes sequential stages of differentiation in cell culture. , 1994, Journal of cell science.

[38]  M. Coroneo,et al.  A Model for Pterygium Formation , 1994, Cornea.

[39]  R. Rice,et al.  Keratinocyte transglutaminase membrane anchorage: analysis of site-directed mutants. , 1993, Biochemistry.

[40]  B. Wieringa,et al.  SKALP/elafin: an elastase inhibitor from cultured human keratinocytes. Purification, cDNA sequence, and evidence for transglutaminase cross-linking. , 1993, The Journal of biological chemistry.

[41]  D. Hohl,et al.  Characterization of human loricrin. Structure and function of a new class of epidermal cell envelope proteins. , 1991, The Journal of biological chemistry.

[42]  L. Lorand,et al.  Labeling of epsilon-lysine crosslinking sites in proteins with peptide substrates of factor XIIIa and transglutaminase. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[43]  S. M. Robinson,et al.  Identification and subcellular distribution of cornified envelope precursor proteins in the transformed human keratinocyte line SV-K14. , 1987, The Journal of investigative dermatology.

[44]  Soo-Youl Kim,et al.  Novel transglutaminase inhibitors reverse the inflammation of allergic conjunctivitis. , 2003, The Journal of clinical investigation.

[45]  R. Eckert,et al.  A regulatory role for p38 delta MAPK in keratinocyte differentiation. Evidence for p38 delta-ERK1/2 complex formation. , 2003, The Journal of biological chemistry.

[46]  P. Steinert,et al.  Small proline-rich proteins are cross-bridging proteins in the cornified cell envelopes of stratified squamous epithelia. , 1998, Journal of structural biology.

[47]  L. Margaritis,et al.  Quantitative changes and ultrastructural alterations of the cornea in response to ultraviolet light. II. Effects of amphibia; elucidation of desmosomal structure and basement membrane synthesis. , 1976, Tissue & cell.