Differentiation-dependent Increases in Lactate Dehydrogenase Activity and Isoenzyme Expression in Rabbit Corneal Epithelial Cells

Hernández-Quintero, Miriam; García-Villegas, Refugio; Castro‐Muñozledo, Federico

doi:10.1006/exer.2001.1110

Cited by 16 publications

(18 citation statements)

References 60 publications

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“…This process proceeds at a steady rate during the lifetime of the tissue and requires that specific gene products be turned on and off in a tightly controlled spatial and temporal manner. Several gene products, including cytokeratins K3 12,13,30 and K12, 27,31 involucrin, 10 and type I transglutaminase, 32 are selectively expressed in the corneal epithelium, and this expression is controlled by specific transcription factors. However, despite this knowledge, information is limited regarding the mechanisms that regulate gene expression in this tissue.…”

Section: Discussionmentioning

confidence: 99%

“…In the corneal epithelium, specific information is available regarding the mechanisms that regulate expression of only a few genes (e.g., cytokeratin genes 12 and the gene encoding lactate dehydrogenase 13 ). Moreover, only limited information is available regarding the mechanisms that regulate involucrin (hINV) gene expression in the corneal epithelium, and regulatory mechanisms have not been investigated in primary human corneal epithelial cell cultures.…”

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confidence: 99%

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Regulation of Involucrin Expression in Normal Human Corneal Epithelial Cells: A Role for Activator Protein One

Adhikary¹,

Crish²,

Lass

et al. 2004

Invest. Ophthalmol. Vis. Sci.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Regulation of Involucrin Expression in Normal Human Corneal Epithelial Cells: A Role for Activator Protein One

Adhikary¹,

Crish²,

Lass

et al. 2004

Invest. Ophthalmol. Vis. Sci.

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“…Differentiation-dependent changes in expression of several genes in the corneal epithelia have been described. 11,43,44 Thus, the genes encoding cytokeratin K3, 2,45 keratin-12, 46 -48 calcium-linked epithelial differentiation protein (CLED protein), 44 transglutaminase type I, 22 and hINV 16 all increase during this progression. However, the mechanisms that regulate gene expression during this process are not well understood.…”

Section: Discussionmentioning

confidence: 96%

“…Lactate dehydrogenase, which is increased during corneal epithelial cell differentiation, is also regulated by Sp1 and AP2 factors, suggesting a shared mode of regulation with cytokeratin K3. 11 Sp1 has also been implicated by cotransfection studies as a suppressor of ␣1-proteinase inhibitor expression in corneal epithelial cells 57 and is involved in regulation of other corneal epithelial genes, including the ␣5-integrin subunit 58 and cytokeratin 4. 59 The ESE-1 transcription factor has been reported to be expressed selectively in differentiated cells and thus may have a role in regulating differentiation.…”

Section: Transcriptional Regulation In the Corneal Epitheliummentioning

confidence: 98%

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An Involucrin Promoter AP1 Transcription Factor Binding Site Is Required for Expression of Involucrin in the Corneal Epithelium In Vivo

Adhikary¹,

Crish²,

Bone³

et al. 2005

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Cell division of corneal limbal stem cells gives rise to transient amplifying cells that ultimately differentiate to form the multilayered corneal epithelium. The mechanisms that regulate changes in gene expression during this process are not well understood. In the present study, the involucrin gene was used as a model to study this regulation. METHODS. Regulation of human involucrin gene expression and promoter activity was assessed using in vivo transgenic mouse models and cultured primary human corneal epithelial cells. RESULTS. Human involucrin (hINV) is a structural protein that is selectively expressed in differentiating corneal epithelial cells. The results reveal that an activator protein one (AP1) DNAbinding site is essential for appropriate basal and stimulusdependent hINV promoter activity. Mutation of this site, AP1-5, results in a loss of hINV gene expression in the corneal epithelium in vivo and in cultured corneal epithelial cells. A gel mobility supershift analysis revealed interaction of the AP1 factors, Fra-1 and JunB, with this element. Inhibition of AP1 function with a dominant-negative form of AP1 also inhibited expression. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C activator, increased hINV gene expression, a response that correlates with increased AP1 factor (Fra-1 and JunB) binding to the hINV gene AP1-5 response element. CONCLUSIONS. These findings point to an essential role for AP1 transcription factors, acting through a distal regulatory region AP1-5 element, in the regulation of involucrin gene expression during corneal epithelial cell differentiation. (Invest Ophthalmol Vis Sci.

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Pax‐6 is expressed early in the differentiation of a corneal epithelial model system

García-Villegas

Escamilla

Sánchez-Guzmán

et al. 2009

Journal Cellular Physiology

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Pax-6 is a regulatory gene with a major role during visual system development, but its association with corneal epithelial differentiation is not clearly established. Using the RCE1-(5T5) cell line, which mimics corneal epithelial differentiation, we analyzed Pax-6 biological role. Immunostaining of proliferating colonies and confluent sheets showed that Pax-6-positive cells were also K3 keratin-positive, suggesting that Pax-6 is expressed in differentiating cells. Pax-6 mRNA was barely expressed in early cell cultures; but after confluence, its levels raised up to fivefold as demonstrated by Northern blot and RT-qPCR. The raise in Pax-6 expression preceded for 9 h the increase in LDH-H and LDH-M mRNAs, previously shown as early markers of corneal epithelial cell differentiation. The full-length mRNAs encoding for the two major Pax-6 isoforms were found at very low levels in proliferating cells, and abundantly expressed in the confluent stratified epithelia; Pax-6 mRNA was 2- to 2.5-fold more abundant than Pax-6(5a) mRNA. The ectopic expression of Pax-6 or Pax-6(5a) decreased proliferative ability leading to the formation of abortive, non-proliferative colonies. In contrast, culture conditions that delay or block corneal epithelial cell differentiation reduced or inhibited the expression of Pax-6. Collectively, results show that Pax-6 is the earlier differentiation marker expressed by corneal epithelial cells, and open the possibility for a major role of Pax-6 as the main driver of the differentiation of corneal epithelial cells.

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Differentiation-dependent Increases in Lactate Dehydrogenase Activity and Isoenzyme Expression in Rabbit Corneal Epithelial Cells

Cited by 16 publications

References 60 publications

Regulation of Involucrin Expression in Normal Human Corneal Epithelial Cells: A Role for Activator Protein One

Regulation of Involucrin Expression in Normal Human Corneal Epithelial Cells: A Role for Activator Protein One

An Involucrin Promoter AP1 Transcription Factor Binding Site Is Required for Expression of Involucrin in the Corneal Epithelium In Vivo

Pax‐6 is expressed early in the differentiation of a corneal epithelial model system

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