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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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Electron microscopy of lacrimal gland. Representative H&E staining and electron microscopy photographs of non-Sjögren syndrome (case 9; A, B, C) and Sjögren syndrome patient (case 4; D, E, F) were shown. Scale bar indicated 5 μm (B and E) and 2 μm (C and F). Structure of lacrimal acinar unit was compact and uniform in non-Sjögren syndrome patients (A and B), but mild acinar atrophy and fibrosis were observed more frequently in Sjögren syndrome patients (D and E). High magnification revealed that structure of mitochondrial cristae (arrows) was severely damaged and swollen in Sjögren syndrome patient (F) compared to that in non-Sjögren syndrome patient (C).
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f3: Electron microscopy of lacrimal gland. Representative H&E staining and electron microscopy photographs of non-Sjögren syndrome (case 9; A, B, C) and Sjögren syndrome patient (case 4; D, E, F) were shown. Scale bar indicated 5 μm (B and E) and 2 μm (C and F). Structure of lacrimal acinar unit was compact and uniform in non-Sjögren syndrome patients (A and B), but mild acinar atrophy and fibrosis were observed more frequently in Sjögren syndrome patients (D and E). High magnification revealed that structure of mitochondrial cristae (arrows) was severely damaged and swollen in Sjögren syndrome patient (F) compared to that in non-Sjögren syndrome patient (C).

Mentions: To determine ultrastructural changes in lacrimal gland, we analyzed samples using electron microscopy. Electron microscopy revealed that the structure of each lacrimal acinar unit was compact and uniform in the non-Sjögren syndrome group. Myoepithelial cells were smooth in shape (Figure 3B). Mild acinar atrophy and fibrosis were observed more frequently in the Sjögren syndrome group (Figure 3E). Infiltration of inflammatory cells was observed in both groups, being particularly marked in the Sjögren syndrome group. High magnification revealed that the structure of mitochondrial cristae was severely damaged and swollen in the Sjögren syndrome group (Figure 3F) compared to that in the non-Sjögren syndrome group (Figure 3C), indicating that mitochondrial damage may be related to Sjögren syndrome.


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)

Electron microscopy of lacrimal gland. Representative H&E staining and electron microscopy photographs of non-Sjögren syndrome (case 9; A, B, C) and Sjögren syndrome patient (case 4; D, E, F) were shown. Scale bar indicated 5 μm (B and E) and 2 μm (C and F). Structure of lacrimal acinar unit was compact and uniform in non-Sjögren syndrome patients (A and B), but mild acinar atrophy and fibrosis were observed more frequently in Sjögren syndrome patients (D and E). High magnification revealed that structure of mitochondrial cristae (arrows) was severely damaged and swollen in Sjögren syndrome patient (F) compared to that in non-Sjögren syndrome patient (C).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Electron microscopy of lacrimal gland. Representative H&E staining and electron microscopy photographs of non-Sjögren syndrome (case 9; A, B, C) and Sjögren syndrome patient (case 4; D, E, F) were shown. Scale bar indicated 5 μm (B and E) and 2 μm (C and F). Structure of lacrimal acinar unit was compact and uniform in non-Sjögren syndrome patients (A and B), but mild acinar atrophy and fibrosis were observed more frequently in Sjögren syndrome patients (D and E). High magnification revealed that structure of mitochondrial cristae (arrows) was severely damaged and swollen in Sjögren syndrome patient (F) compared to that in non-Sjögren syndrome patient (C).
Mentions: To determine ultrastructural changes in lacrimal gland, we analyzed samples using electron microscopy. Electron microscopy revealed that the structure of each lacrimal acinar unit was compact and uniform in the non-Sjögren syndrome group. Myoepithelial cells were smooth in shape (Figure 3B). Mild acinar atrophy and fibrosis were observed more frequently in the Sjögren syndrome group (Figure 3E). Infiltration of inflammatory cells was observed in both groups, being particularly marked in the Sjögren syndrome group. High magnification revealed that the structure of mitochondrial cristae was severely damaged and swollen in the Sjögren syndrome group (Figure 3F) compared to that in the non-Sjögren syndrome group (Figure 3C), indicating that mitochondrial damage may be related to Sjögren syndrome.

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