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Gene expression profile of the skin in the 'hairpoor' (HrHp) mice by microarray analysis.

Kim BK, Baek IC, Lee HY, Kim JK, Song HH, Yoon SK - BMC Genomics (2010)

Bottom Line: Among the 1861 genes with a > 1.2-fold increase in expression, further analysis showed that the expression of eight genes known to have a close relationship with hair follicle development, ascertained by conducting real-time PCR on skin RNA produced during hair follicle morphogenesis (P0-P14), indicated that four genes, Wif1, Casp14, Krt71, and Sfrp1, showed a consistent expression pattern with respect to HR overexpression in vivo.Wif1 and Casp14 were found to be upregulated, whereas Krt71 and Sfrp1 were downregulated in cells overexpressing HR in transient transfection experiments on keratinocytes, suggesting that HR may transcriptionally regulate these genes.Further studies are required to understand the mechanism of this regulation by the HR cofactor.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Sciences, The Catholic University of Korea, 505 Banpo-dong, Seoul 137-701, Korea.

ABSTRACT

Background: The transcriptional cofactor, Hairless (HR), acts as one of the key regulators of hair follicle cycling; the loss of function mutations is the cause of the expression of the hairless phenotype in humans and mice. Recently, we reported a new Hr mutant mouse called 'Hairpoor' (Hr(Hp)). These mutants harbor a gain of the function mutation, T403A, in the Hr gene. This confers the overexpression of HR and Hr(Hp) is an animal model of Marie Unna hereditary hypotrichosis in humans. In the present study, the expression profile of Hr(Hp)/Hr(Hp) skin was investigated using microarray analysis to identify genes whose expression was affected by the overexpression of HR.

Results: From 45,282 mouse probes, differential expressions in 43 (>2-fold), 306 (>1.5-fold), and 1861 genes (>1.2-fold) in skin from Hr(Hp)/Hr(Hp) mice were discovered and compared with skin from wild-type mice. Among the 1861 genes with a > 1.2-fold increase in expression, further analysis showed that the expression of eight genes known to have a close relationship with hair follicle development, ascertained by conducting real-time PCR on skin RNA produced during hair follicle morphogenesis (P0-P14), indicated that four genes, Wif1, Casp14, Krt71, and Sfrp1, showed a consistent expression pattern with respect to HR overexpression in vivo.

Conclusion: Wif1 and Casp14 were found to be upregulated, whereas Krt71 and Sfrp1 were downregulated in cells overexpressing HR in transient transfection experiments on keratinocytes, suggesting that HR may transcriptionally regulate these genes. Further studies are required to understand the mechanism of this regulation by the HR cofactor.

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Changes in expression of Wif1, Sfrp1, Casp14, and Krt71 in Hr-transfected PAM212 cells. (A) Expression of Wif1, Sfrp1, Casp14, and Krt71in normal PAM212 cells using RT-PCR. M: size marker. (B) Western blot analysis showing the HR protein expressed in Hr-transfected PAM212 cells. β-tubulin indicates equal amount of protein loading. (C) Regulation of Wif1, Sfrp1, Casp14, and Krt71 gene expression by HR in Hr-transfected PAM212 cells by real-time PCR. The Y-axis indicates the fold difference in relative expression levels of each gene in HR-overexpressing cells compared with controls. *p value < 0.05. Results are the average of three independent experiments conducted in duplicate.
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Figure 4: Changes in expression of Wif1, Sfrp1, Casp14, and Krt71 in Hr-transfected PAM212 cells. (A) Expression of Wif1, Sfrp1, Casp14, and Krt71in normal PAM212 cells using RT-PCR. M: size marker. (B) Western blot analysis showing the HR protein expressed in Hr-transfected PAM212 cells. β-tubulin indicates equal amount of protein loading. (C) Regulation of Wif1, Sfrp1, Casp14, and Krt71 gene expression by HR in Hr-transfected PAM212 cells by real-time PCR. The Y-axis indicates the fold difference in relative expression levels of each gene in HR-overexpressing cells compared with controls. *p value < 0.05. Results are the average of three independent experiments conducted in duplicate.

Mentions: To further analyze whether the expressions of Wif1, Sfrp1, Casp14, and Krt71 were directly regulated by HR, we investigated changes in expression of these genes in the presence of overexpressed HR in a transient expression system using the mouse keratinocyte cell line, PAM212. RT-PCR revealed all the genes normally expressed in PAM212 cells (Figure 4A); transfection of PAM212 cells with Hr cDNA construct resulted in expression of HR (Figure 4B). The expression of all four genes was affected by the presence of HR: expression of Wif1 and Casp14 was increased 1.85- and 1.57-fold in HR-overexpressed cells compared to the mock-transfected PAM212 cells, respectively. In contrast, the relative expression of Sfrp1 and Krt71 was decreased to 0.61- and 0.52-fold (Figure 4C) in HR-expressing cells compared with control cells. These results were consistent with their expression pattern in vivo and strongly suggested that HR may directly regulate expression of these genes.


Gene expression profile of the skin in the 'hairpoor' (HrHp) mice by microarray analysis.

Kim BK, Baek IC, Lee HY, Kim JK, Song HH, Yoon SK - BMC Genomics (2010)

Changes in expression of Wif1, Sfrp1, Casp14, and Krt71 in Hr-transfected PAM212 cells. (A) Expression of Wif1, Sfrp1, Casp14, and Krt71in normal PAM212 cells using RT-PCR. M: size marker. (B) Western blot analysis showing the HR protein expressed in Hr-transfected PAM212 cells. β-tubulin indicates equal amount of protein loading. (C) Regulation of Wif1, Sfrp1, Casp14, and Krt71 gene expression by HR in Hr-transfected PAM212 cells by real-time PCR. The Y-axis indicates the fold difference in relative expression levels of each gene in HR-overexpressing cells compared with controls. *p value < 0.05. Results are the average of three independent experiments conducted in duplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3091768&req=5

Figure 4: Changes in expression of Wif1, Sfrp1, Casp14, and Krt71 in Hr-transfected PAM212 cells. (A) Expression of Wif1, Sfrp1, Casp14, and Krt71in normal PAM212 cells using RT-PCR. M: size marker. (B) Western blot analysis showing the HR protein expressed in Hr-transfected PAM212 cells. β-tubulin indicates equal amount of protein loading. (C) Regulation of Wif1, Sfrp1, Casp14, and Krt71 gene expression by HR in Hr-transfected PAM212 cells by real-time PCR. The Y-axis indicates the fold difference in relative expression levels of each gene in HR-overexpressing cells compared with controls. *p value < 0.05. Results are the average of three independent experiments conducted in duplicate.
Mentions: To further analyze whether the expressions of Wif1, Sfrp1, Casp14, and Krt71 were directly regulated by HR, we investigated changes in expression of these genes in the presence of overexpressed HR in a transient expression system using the mouse keratinocyte cell line, PAM212. RT-PCR revealed all the genes normally expressed in PAM212 cells (Figure 4A); transfection of PAM212 cells with Hr cDNA construct resulted in expression of HR (Figure 4B). The expression of all four genes was affected by the presence of HR: expression of Wif1 and Casp14 was increased 1.85- and 1.57-fold in HR-overexpressed cells compared to the mock-transfected PAM212 cells, respectively. In contrast, the relative expression of Sfrp1 and Krt71 was decreased to 0.61- and 0.52-fold (Figure 4C) in HR-expressing cells compared with control cells. These results were consistent with their expression pattern in vivo and strongly suggested that HR may directly regulate expression of these genes.

Bottom Line: Among the 1861 genes with a > 1.2-fold increase in expression, further analysis showed that the expression of eight genes known to have a close relationship with hair follicle development, ascertained by conducting real-time PCR on skin RNA produced during hair follicle morphogenesis (P0-P14), indicated that four genes, Wif1, Casp14, Krt71, and Sfrp1, showed a consistent expression pattern with respect to HR overexpression in vivo.Wif1 and Casp14 were found to be upregulated, whereas Krt71 and Sfrp1 were downregulated in cells overexpressing HR in transient transfection experiments on keratinocytes, suggesting that HR may transcriptionally regulate these genes.Further studies are required to understand the mechanism of this regulation by the HR cofactor.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Sciences, The Catholic University of Korea, 505 Banpo-dong, Seoul 137-701, Korea.

ABSTRACT

Background: The transcriptional cofactor, Hairless (HR), acts as one of the key regulators of hair follicle cycling; the loss of function mutations is the cause of the expression of the hairless phenotype in humans and mice. Recently, we reported a new Hr mutant mouse called 'Hairpoor' (Hr(Hp)). These mutants harbor a gain of the function mutation, T403A, in the Hr gene. This confers the overexpression of HR and Hr(Hp) is an animal model of Marie Unna hereditary hypotrichosis in humans. In the present study, the expression profile of Hr(Hp)/Hr(Hp) skin was investigated using microarray analysis to identify genes whose expression was affected by the overexpression of HR.

Results: From 45,282 mouse probes, differential expressions in 43 (>2-fold), 306 (>1.5-fold), and 1861 genes (>1.2-fold) in skin from Hr(Hp)/Hr(Hp) mice were discovered and compared with skin from wild-type mice. Among the 1861 genes with a > 1.2-fold increase in expression, further analysis showed that the expression of eight genes known to have a close relationship with hair follicle development, ascertained by conducting real-time PCR on skin RNA produced during hair follicle morphogenesis (P0-P14), indicated that four genes, Wif1, Casp14, Krt71, and Sfrp1, showed a consistent expression pattern with respect to HR overexpression in vivo.

Conclusion: Wif1 and Casp14 were found to be upregulated, whereas Krt71 and Sfrp1 were downregulated in cells overexpressing HR in transient transfection experiments on keratinocytes, suggesting that HR may transcriptionally regulate these genes. Further studies are required to understand the mechanism of this regulation by the HR cofactor.

Show MeSH
Related in: MedlinePlus