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ADAM and ADAMTS gene expression in native and wound healing human lens epithelial cells.

Hodgkinson LM, Wang L, Duncan G, Edwards DR, Wormstone IM - Mol. Vis. (2010)

Bottom Line: Data were analyzed in terms of cycle threshold number (C(T)) and also normalized relative to endogenous 18S rRNA.High expression of ADAM-9, -10, -15, and -17 was detected in all native lens regions.ADAM-15 expression was a characteristic of the native lens epithelia more than the fibers.

View Article: PubMed Central - PubMed

Affiliation: School of Biological Sciences, University of East Anglia, Norwich, UK. lisa.hodg@hotmail.com

ABSTRACT

Purpose: The ADAMs (a disintegrin and metalloproteinase) and the ADAMTSs (a disintegrin and metalloproteinase with thrombospondin-like motifs) are extracellular proteases that mediate cellular interactions and cell signaling via the modulation of adhesion and the cleavage of cell surface protein ectodomains and extracellular matrix molecules. Gene expression profiling was undertaken to better understand the role of the ADAM and ADAMTS families in the clear native human lenses and following surgical injury with particular relevance to posterior capsule opacification.

Methods: To carry out profile analysis, the lens (t=0d) was dissected into three regions; anterior epithelia, equatorial region, and fiber cells. Capsular bag culture was undertaken as a means of assessing short-term changes (t=6d) and post-cataractous lens capsular bags (ex vivo) were used to predict long-term changes in ADAM/ADAMTS gene expression. RNA was isolated and quantitative real-time (TaqMan) reverse transcription-PCR (RT-PCR) performed. Data were analyzed in terms of cycle threshold number (C(T)) and also normalized relative to endogenous 18S rRNA.

Results: High expression of ADAM-9, -10, -15, and -17 was detected in all native lens regions. ADAM-15 expression was a characteristic of the native lens epithelia more than the fibers. Post-surgical injury, lens capsular bags showed a positive shift in ADAM/ADAMTS expression that was significant for ADAM-9, -15, and ADAMTS-3. Ex vivo capsular bags, with a long-term post surgical injury period, maintained high expression of ADAM-9 and -10 genes.

Conclusions: The native human lens expresses ADAM and ADAMTS genes that are differentially regulated following surgical injury. Roles in maintaining cellular adhesion may be of particular importance to native lens tissue integrity and may be lost in the lens wound healing response following cataract surgery.

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Related in: MedlinePlus

ADAM/ADAMTS gene expression in response to mechanical trauma. Gene expression profiles were classified in CBs; at t=0 following in vitro sham cataract operation (left panel), at t=6d, following short-term culture (central panel) and ex vivo after cataract surgery (right panel). The cycle threshold number (CT) was used to classify gene expression as either very high (CT≤20), high (CT=21–25), moderate (CT=26–30), low (CT=31–38), or negligible to undetected (CT=39–40). Donor information; category (t=0, 6d, ex vivo), sex (F; female, M; male), age and ophthalmic history. t=0: F, 74, none; M, 83, none; M, 81, none. t=6d; M, 76, none; F, 76, none; M, 69, no info. Ex vivo; M, 75, bilateral cataract surgery 2001; F, 76, no info; M, 70, no info.
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f3: ADAM/ADAMTS gene expression in response to mechanical trauma. Gene expression profiles were classified in CBs; at t=0 following in vitro sham cataract operation (left panel), at t=6d, following short-term culture (central panel) and ex vivo after cataract surgery (right panel). The cycle threshold number (CT) was used to classify gene expression as either very high (CT≤20), high (CT=21–25), moderate (CT=26–30), low (CT=31–38), or negligible to undetected (CT=39–40). Donor information; category (t=0, 6d, ex vivo), sex (F; female, M; male), age and ophthalmic history. t=0: F, 74, none; M, 83, none; M, 81, none. t=6d; M, 76, none; F, 76, none; M, 69, no info. Ex vivo; M, 75, bilateral cataract surgery 2001; F, 76, no info; M, 70, no info.

Mentions: In the qualitative analysis of the raw data cycle threshold (CT) value, before 18S normalization (Figure 3), ADAM-12, -19, and ADAMTS-14 showed a positive shift in gene expression during short-term culture (t=6d) relative to t=0. The expression of ADAM-12 and -19 increased from low to moderate and ADAMTS-14, which was negligible/undetected before culture, increased to a low expression level. The classification of the remaining ADAM/ADAMTS genes expressed (ADAM-8, -9, -10, -15, -17, -28, ADAMTS-1, -2, and -3) did not change with short-term culture relative to t=0.


ADAM and ADAMTS gene expression in native and wound healing human lens epithelial cells.

Hodgkinson LM, Wang L, Duncan G, Edwards DR, Wormstone IM - Mol. Vis. (2010)

ADAM/ADAMTS gene expression in response to mechanical trauma. Gene expression profiles were classified in CBs; at t=0 following in vitro sham cataract operation (left panel), at t=6d, following short-term culture (central panel) and ex vivo after cataract surgery (right panel). The cycle threshold number (CT) was used to classify gene expression as either very high (CT≤20), high (CT=21–25), moderate (CT=26–30), low (CT=31–38), or negligible to undetected (CT=39–40). Donor information; category (t=0, 6d, ex vivo), sex (F; female, M; male), age and ophthalmic history. t=0: F, 74, none; M, 83, none; M, 81, none. t=6d; M, 76, none; F, 76, none; M, 69, no info. Ex vivo; M, 75, bilateral cataract surgery 2001; F, 76, no info; M, 70, no info.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3008711&req=5

f3: ADAM/ADAMTS gene expression in response to mechanical trauma. Gene expression profiles were classified in CBs; at t=0 following in vitro sham cataract operation (left panel), at t=6d, following short-term culture (central panel) and ex vivo after cataract surgery (right panel). The cycle threshold number (CT) was used to classify gene expression as either very high (CT≤20), high (CT=21–25), moderate (CT=26–30), low (CT=31–38), or negligible to undetected (CT=39–40). Donor information; category (t=0, 6d, ex vivo), sex (F; female, M; male), age and ophthalmic history. t=0: F, 74, none; M, 83, none; M, 81, none. t=6d; M, 76, none; F, 76, none; M, 69, no info. Ex vivo; M, 75, bilateral cataract surgery 2001; F, 76, no info; M, 70, no info.
Mentions: In the qualitative analysis of the raw data cycle threshold (CT) value, before 18S normalization (Figure 3), ADAM-12, -19, and ADAMTS-14 showed a positive shift in gene expression during short-term culture (t=6d) relative to t=0. The expression of ADAM-12 and -19 increased from low to moderate and ADAMTS-14, which was negligible/undetected before culture, increased to a low expression level. The classification of the remaining ADAM/ADAMTS genes expressed (ADAM-8, -9, -10, -15, -17, -28, ADAMTS-1, -2, and -3) did not change with short-term culture relative to t=0.

Bottom Line: Data were analyzed in terms of cycle threshold number (C(T)) and also normalized relative to endogenous 18S rRNA.High expression of ADAM-9, -10, -15, and -17 was detected in all native lens regions.ADAM-15 expression was a characteristic of the native lens epithelia more than the fibers.

View Article: PubMed Central - PubMed

Affiliation: School of Biological Sciences, University of East Anglia, Norwich, UK. lisa.hodg@hotmail.com

ABSTRACT

Purpose: The ADAMs (a disintegrin and metalloproteinase) and the ADAMTSs (a disintegrin and metalloproteinase with thrombospondin-like motifs) are extracellular proteases that mediate cellular interactions and cell signaling via the modulation of adhesion and the cleavage of cell surface protein ectodomains and extracellular matrix molecules. Gene expression profiling was undertaken to better understand the role of the ADAM and ADAMTS families in the clear native human lenses and following surgical injury with particular relevance to posterior capsule opacification.

Methods: To carry out profile analysis, the lens (t=0d) was dissected into three regions; anterior epithelia, equatorial region, and fiber cells. Capsular bag culture was undertaken as a means of assessing short-term changes (t=6d) and post-cataractous lens capsular bags (ex vivo) were used to predict long-term changes in ADAM/ADAMTS gene expression. RNA was isolated and quantitative real-time (TaqMan) reverse transcription-PCR (RT-PCR) performed. Data were analyzed in terms of cycle threshold number (C(T)) and also normalized relative to endogenous 18S rRNA.

Results: High expression of ADAM-9, -10, -15, and -17 was detected in all native lens regions. ADAM-15 expression was a characteristic of the native lens epithelia more than the fibers. Post-surgical injury, lens capsular bags showed a positive shift in ADAM/ADAMTS expression that was significant for ADAM-9, -15, and ADAMTS-3. Ex vivo capsular bags, with a long-term post surgical injury period, maintained high expression of ADAM-9 and -10 genes.

Conclusions: The native human lens expresses ADAM and ADAMTS genes that are differentially regulated following surgical injury. Roles in maintaining cellular adhesion may be of particular importance to native lens tissue integrity and may be lost in the lens wound healing response following cataract surgery.

Show MeSH
Related in: MedlinePlus