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Comparison of telomere length and association with progenitor cell markers in lacrimal gland between Sjögren syndrome and non-Sjögren syndrome dry eye patients.

Kawashima M, Kawakita T, Maida Y, Kamoi M, Ogawa Y, Shimmura S, Masutomi K, Tsubota K - Mol. Vis. (2011)

Bottom Line: Immunostaining for p63, nucleostemin, ATP-binding cassette, sub-family G, member 2 (ABCG2), and nestin was also performed.Among the samples from the non-Sjögren syndrome group, immunostaining revealed that p63 was expressed in 1-3 acinar cells in each acinar unit and continuously in the basal layer of duct cells.The results of this study indicate that 1) telo-FISH is a viable method of assessing telomere length in lacrimal gland, and 2) telomere length in Sjögren syndrome is shorter and associated with lower levels of expression of p63 and nucleostemin than in non-Sjögren syndrome.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.

ABSTRACT

Purpose: Indicators of aging such as disruption of telomeric function due to shortening may be more frequent in dysfunctional lacrimal gland. The aims of this study were to 1) determine the viability of quantitative fluorescence in situ hybridization of telomeres (telo-FISH) for the assessment of telomere length in lacrimal gland in Sjögren and non- Sjögren syndrome patients; and 2) investigate the relationship between progenitor cell markers and telomere length in both groups.

Methods: Quantitative fluorescence in situ hybridization with a peptide nucleic acid probe complementary to the telomere repeat sequence was performed on frozen sections from human lacrimal gland tissues. The mean fluorescence intensity of telomere spots was automatically quantified by image analysis as relative telomere length in lacrimal gland epithelial cells. Immunostaining for p63, nucleostemin, ATP-binding cassette, sub-family G, member 2 (ABCG2), and nestin was also performed.

Results: Telomere intensity in the Sjögren syndrome group (6,785.0±455) was significantly lower than that in the non-Sjögren syndrome group (7,494.7±477; p=0.02). Among the samples from the non-Sjögren syndrome group, immunostaining revealed that p63 was expressed in 1-3 acinar cells in each acinar unit and continuously in the basal layer of duct cells. In contrast, in the Sjögren syndrome group, p63 and nucleostemin showed a lower level of expression. ABCG2 was expressed in acinar cells in both sjogren and non-Sjogren syndrome.

Conclusions: The results of this study indicate that 1) telo-FISH is a viable method of assessing telomere length in lacrimal gland, and 2) telomere length in Sjögren syndrome is shorter and associated with lower levels of expression of p63 and nucleostemin than in non-Sjögren syndrome.

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Immunostaining for progenitor markers. In non-Sjögren syndrome, p63 (red) was expressed in 2–4 cells in each acinar unit (A) and all ductal basal cells (B; case 7). In Sjögren syndrome, p63 was weakly expressed with irregular pattern. (case 2; F, G). Nucleostemin was expressed with a similar pattern in non-Sjögren syndrome (case 7; C) and Sjögren syndrome (case 2; H). Nuclei were counterstained with DAPI (blue). ABCG2 (red) was expressed in intercellular junction and cytoplasm in acinar unit (F, I), and weaker in Sjögren syndrome. Nestin was expressed strongly in some location in Sjögren syndrome (E, J). Scale bars indicate 50 μm (A-C, E-H, J) and 20 μm (D, I), SS=Sjögren syndrome, non-SS=non Sjögren syndrome.
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f2: Immunostaining for progenitor markers. In non-Sjögren syndrome, p63 (red) was expressed in 2–4 cells in each acinar unit (A) and all ductal basal cells (B; case 7). In Sjögren syndrome, p63 was weakly expressed with irregular pattern. (case 2; F, G). Nucleostemin was expressed with a similar pattern in non-Sjögren syndrome (case 7; C) and Sjögren syndrome (case 2; H). Nuclei were counterstained with DAPI (blue). ABCG2 (red) was expressed in intercellular junction and cytoplasm in acinar unit (F, I), and weaker in Sjögren syndrome. Nestin was expressed strongly in some location in Sjögren syndrome (E, J). Scale bars indicate 50 μm (A-C, E-H, J) and 20 μm (D, I), SS=Sjögren syndrome, non-SS=non Sjögren syndrome.

Mentions: To investigate the relationship between telomere length and progenitor cell markers, we performed immunostaining for p63 and nucleostemin. In the non-Sjögren syndrome group, p63 was expressed in 2–4 acinar cells in each acinar unit (Figure 2A) and in the basal layer of duct basal cells continuously (Figure 2B). In contrast, p63 showed a lower level of expression and was only sparsely expressed in the basal layer of duct cells in the Sjögren syndrome group (Figure 2F,G). Nucleostemin showed a similar pattern of expression as p63, being higher and more regularly expressed in the non-Sjögren syndrome group than in the Sjögren syndrome group (Figure 2C,H). ABCG2 was expressed in intercellular junction and cytoplasm of most acinar unit regularly in the non-Sjögren syndrome group (Figure 2D). Whereas acinar unit was decreased in the Sjögren syndrome group, and acinar unit with ABCG2 expression was also decreased (Figure 2I). Nestin was expressed outside the acinar unit in both group, Elongated cells with nestin expression were observed more frequently in the SS group, compared with the non-SS group. Those nestin-positive cells were not observed uniformly in all locations, but formed cell clusters at tissue damaged areas in the SS group (Figure 2F,J). Telomere length was shorter and expression of progenitor markers weaker or non-existent in the Sjögren syndrome group.


Comparison of telomere length and association with progenitor cell markers in lacrimal gland between Sjögren syndrome and non-Sjögren syndrome dry eye patients.

Kawashima M, Kawakita T, Maida Y, Kamoi M, Ogawa Y, Shimmura S, Masutomi K, Tsubota K - Mol. Vis. (2011)

Immunostaining for progenitor markers. In non-Sjögren syndrome, p63 (red) was expressed in 2–4 cells in each acinar unit (A) and all ductal basal cells (B; case 7). In Sjögren syndrome, p63 was weakly expressed with irregular pattern. (case 2; F, G). Nucleostemin was expressed with a similar pattern in non-Sjögren syndrome (case 7; C) and Sjögren syndrome (case 2; H). Nuclei were counterstained with DAPI (blue). ABCG2 (red) was expressed in intercellular junction and cytoplasm in acinar unit (F, I), and weaker in Sjögren syndrome. Nestin was expressed strongly in some location in Sjögren syndrome (E, J). Scale bars indicate 50 μm (A-C, E-H, J) and 20 μm (D, I), SS=Sjögren syndrome, non-SS=non Sjögren syndrome.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108899&req=5

f2: Immunostaining for progenitor markers. In non-Sjögren syndrome, p63 (red) was expressed in 2–4 cells in each acinar unit (A) and all ductal basal cells (B; case 7). In Sjögren syndrome, p63 was weakly expressed with irregular pattern. (case 2; F, G). Nucleostemin was expressed with a similar pattern in non-Sjögren syndrome (case 7; C) and Sjögren syndrome (case 2; H). Nuclei were counterstained with DAPI (blue). ABCG2 (red) was expressed in intercellular junction and cytoplasm in acinar unit (F, I), and weaker in Sjögren syndrome. Nestin was expressed strongly in some location in Sjögren syndrome (E, J). Scale bars indicate 50 μm (A-C, E-H, J) and 20 μm (D, I), SS=Sjögren syndrome, non-SS=non Sjögren syndrome.
Mentions: To investigate the relationship between telomere length and progenitor cell markers, we performed immunostaining for p63 and nucleostemin. In the non-Sjögren syndrome group, p63 was expressed in 2–4 acinar cells in each acinar unit (Figure 2A) and in the basal layer of duct basal cells continuously (Figure 2B). In contrast, p63 showed a lower level of expression and was only sparsely expressed in the basal layer of duct cells in the Sjögren syndrome group (Figure 2F,G). Nucleostemin showed a similar pattern of expression as p63, being higher and more regularly expressed in the non-Sjögren syndrome group than in the Sjögren syndrome group (Figure 2C,H). ABCG2 was expressed in intercellular junction and cytoplasm of most acinar unit regularly in the non-Sjögren syndrome group (Figure 2D). Whereas acinar unit was decreased in the Sjögren syndrome group, and acinar unit with ABCG2 expression was also decreased (Figure 2I). Nestin was expressed outside the acinar unit in both group, Elongated cells with nestin expression were observed more frequently in the SS group, compared with the non-SS group. Those nestin-positive cells were not observed uniformly in all locations, but formed cell clusters at tissue damaged areas in the SS group (Figure 2F,J). Telomere length was shorter and expression of progenitor markers weaker or non-existent in the Sjögren syndrome group.

Bottom Line: Immunostaining for p63, nucleostemin, ATP-binding cassette, sub-family G, member 2 (ABCG2), and nestin was also performed.Among the samples from the non-Sjögren syndrome group, immunostaining revealed that p63 was expressed in 1-3 acinar cells in each acinar unit and continuously in the basal layer of duct cells.The results of this study indicate that 1) telo-FISH is a viable method of assessing telomere length in lacrimal gland, and 2) telomere length in Sjögren syndrome is shorter and associated with lower levels of expression of p63 and nucleostemin than in non-Sjögren syndrome.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.

ABSTRACT

Purpose: Indicators of aging such as disruption of telomeric function due to shortening may be more frequent in dysfunctional lacrimal gland. The aims of this study were to 1) determine the viability of quantitative fluorescence in situ hybridization of telomeres (telo-FISH) for the assessment of telomere length in lacrimal gland in Sjögren and non- Sjögren syndrome patients; and 2) investigate the relationship between progenitor cell markers and telomere length in both groups.

Methods: Quantitative fluorescence in situ hybridization with a peptide nucleic acid probe complementary to the telomere repeat sequence was performed on frozen sections from human lacrimal gland tissues. The mean fluorescence intensity of telomere spots was automatically quantified by image analysis as relative telomere length in lacrimal gland epithelial cells. Immunostaining for p63, nucleostemin, ATP-binding cassette, sub-family G, member 2 (ABCG2), and nestin was also performed.

Results: Telomere intensity in the Sjögren syndrome group (6,785.0±455) was significantly lower than that in the non-Sjögren syndrome group (7,494.7±477; p=0.02). Among the samples from the non-Sjögren syndrome group, immunostaining revealed that p63 was expressed in 1-3 acinar cells in each acinar unit and continuously in the basal layer of duct cells. In contrast, in the Sjögren syndrome group, p63 and nucleostemin showed a lower level of expression. ABCG2 was expressed in acinar cells in both sjogren and non-Sjogren syndrome.

Conclusions: The results of this study indicate that 1) telo-FISH is a viable method of assessing telomere length in lacrimal gland, and 2) telomere length in Sjögren syndrome is shorter and associated with lower levels of expression of p63 and nucleostemin than in non-Sjögren syndrome.

Show MeSH
Related in: MedlinePlus