PURPOSE: HtrA1 belongs to the high temperature requirement factor A family of serine proteases, which are involved in protein quality control and cell fate. A single-nucleotide polymorphism (SNP), rs11200638, in the promoter of HtrA1 at chromosome 10q26 is reported as a likely causal variant for age-related macular degeneration (AMD). The SNP is located in the regulatory region and increases production of HtrA1 protein. This study investigates HtrA1 expression and SNP genotypes in archived ocular slides with AMD. METHODS: Macular, nonretinal, and peripheral retinal cells were microdissected from archived slides from 57 eyes with AMD and 16 age-matched, non-AMD controls. HtrA1 rs11200638 SNP genotyping was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis. HtrA1 transcripts were measured using real-time reverse transcriptase-PCR. HtrA1 protein expression was evaluated using avidin-biotin complex immunohistochemistry. RESULTS: HtrA1 (G/A) SNP was successfully genotyped in 52 AMD cases and 13 non-AMD subjects. The frequencies of the risk allele (A) were 55 of 104 (52.9%) and 8 of 26 (30.8%) in AMD and control groups, respectively. HtrA1 mRNA was detected in normal peripheral and macular retinas, higher in the periphery than maculae. HtrA1 mRNA was much higher in the macula and a lot lower in the periphery of the AMD eyes as compared to control eyes. HtrA1 protein was expressed in normal retinal vascular endothelia and retinal pigment epithelia. Intense immunoreaction against HtrA1 was found in AMD lesions, slightly more in wet than dry AMD lesions. CONCLUSION: This study successfully analyzes HtrA1 SNP and transcript expression in microdissected cells from archived paraffin fixed slides. Up-regulation of HtrA1 is detected in the macular lesions of AMD eyes. The data further suggest that rs11200638 in HtrA1 promoter is associated with AMD development.
PURPOSE:HtrA1 belongs to the high temperature requirement factor A family of serine proteases, which are involved in protein quality control and cell fate. A single-nucleotide polymorphism (SNP), rs11200638, in the promoter of HtrA1 at chromosome 10q26 is reported as a likely causal variant for age-related macular degeneration (AMD). The SNP is located in the regulatory region and increases production of HtrA1 protein. This study investigates HtrA1 expression and SNP genotypes in archived ocular slides with AMD. METHODS: Macular, nonretinal, and peripheral retinal cells were microdissected from archived slides from 57 eyes with AMD and 16 age-matched, non-AMD controls. HtrA1rs11200638 SNP genotyping was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis. HtrA1 transcripts were measured using real-time reverse transcriptase-PCR. HtrA1 protein expression was evaluated using avidin-biotin complex immunohistochemistry. RESULTS:HtrA1 (G/A) SNP was successfully genotyped in 52 AMD cases and 13 non-AMD subjects. The frequencies of the risk allele (A) were 55 of 104 (52.9%) and 8 of 26 (30.8%) in AMD and control groups, respectively. HtrA1 mRNA was detected in normal peripheral and macular retinas, higher in the periphery than maculae. HtrA1 mRNA was much higher in the macula and a lot lower in the periphery of the AMD eyes as compared to control eyes. HtrA1 protein was expressed in normal retinal vascular endothelia and retinal pigment epithelia. Intense immunoreaction against HtrA1 was found in AMD lesions, slightly more in wet than dry AMD lesions. CONCLUSION: This study successfully analyzes HtrA1 SNP and transcript expression in microdissected cells from archived paraffin fixed slides. Up-regulation of HtrA1 is detected in the macular lesions of AMD eyes. The data further suggest that rs11200638 in HtrA1 promoter is associated with AMD development.
Authors: Jingsheng Tuo; Baitang Ning; Christine M Bojanowski; Zhong-Ning Lin; Robert J Ross; George F Reed; Defen Shen; Xiaodong Jiao; Min Zhou; Emily Y Chew; Fred F Kadlubar; Chi-Chao Chan Journal: Proc Natl Acad Sci U S A Date: 2006-06-05 Impact factor: 11.205
Authors: Yvette P Conley; Johanna Jakobsdottir; Tammy Mah; Daniel E Weeks; Ronald Klein; Lewis Kuller; Robert E Ferrell; Michael B Gorin Journal: Hum Mol Genet Date: 2006-09-25 Impact factor: 6.150
Authors: Julian Maller; Sarah George; Shaun Purcell; Jes Fagerness; David Altshuler; Mark J Daly; Johanna M Seddon Journal: Nat Genet Date: 2006-08-27 Impact factor: 38.330
Authors: Silke Schmidt; Michael A Hauser; William K Scott; Eric A Postel; Anita Agarwal; Paul Gallins; Frank Wong; Yu Sarah Chen; Kylee Spencer; Nathalie Schnetz-Boutaud; Jonathan L Haines; Margaret A Pericak-Vance Journal: Am J Hum Genet Date: 2006-03-20 Impact factor: 11.025
Authors: Andrew Dewan; Mugen Liu; Stephen Hartman; Samuel Shao-Min Zhang; David T L Liu; Connie Zhao; Pancy O S Tam; Wai Man Chan; Dennis S C Lam; Michael Snyder; Colin Barnstable; Chi Pui Pang; Josephine Hoh Journal: Science Date: 2006-10-19 Impact factor: 47.728
Authors: Andrea Rivera; Sheila A Fisher; Lars G Fritsche; Claudia N Keilhauer; Peter Lichtner; Thomas Meitinger; Bernhard H F Weber Journal: Hum Mol Genet Date: 2005-09-20 Impact factor: 6.150
Authors: Zhenglin Yang; Nicola J Camp; Hui Sun; Zongzhong Tong; Daniel Gibbs; D Joshua Cameron; Haoyu Chen; Yu Zhao; Erik Pearson; Xi Li; Jeremy Chien; Andrew Dewan; Jennifer Harmon; Paul S Bernstein; Viji Shridhar; Norman A Zabriskie; Josephine Hoh; Kimberly Howes; Kang Zhang Journal: Science Date: 2006-10-19 Impact factor: 47.728
Authors: S Scott Whitmore; Elliott H Sohn; Kathleen R Chirco; Arlene V Drack; Edwin M Stone; Budd A Tucker; Robert F Mullins Journal: Prog Retin Eye Res Date: 2014-12-05 Impact factor: 21.198