A S Jun1, S H Liu, E H Koo, D V Do, W J Stark, J D Gottsch. 1. Cornea and External Disease Division, Wilmer Ophthalmological Institute, The Johns Hopkins Medical Institutions, Maumenee 317, 600 N Wolfe St, Baltimore, MD 21287, USA.
Abstract
OBJECTIVES: To use microarray analysis to identify genes expressed in human donor corneas and to create a preliminary, comprehensive database of human corneal gene expression. METHODS: A complementary DNA (cDNA) library was constructed from transplant-quality, human donor corneas. Biotin-labeled RNA was transcribed from the cDNA library and hybridized in duplicate to microarrays containing approximately 5600 human genes. Results were analyzed using a gene database of the National Institutes of Health, Bethesda, Md. Reverse transcriptase polymerase chain reaction analysis was performed to confirm corneal expression of genes identified by microarray analysis. RESULTS: Duplicate microarrays identified the expression of 1200 genes in human donor corneas. Chromosomal loci had been assigned to 1025 (85%) of these genes. A preliminary database of human corneal gene expression was compiled. A Web site containing these genes was created. Six collagen genes were identified that had not previously been localized within the cornea. Five apoptosis-related genes were identified, 4 of which had not previously been localized within the cornea. Three genes previously shown to cause corneal diseases were identified. Reverse transcriptase polymerase chain reaction analysis of genes identified by microarray analysis confirmed the corneal expression of 2 apoptosis-related genes and 1 collagen gene. CONCLUSIONS: Microarray analysis of healthy human donor corneas has produced a preliminary, comprehensive database of corneal gene expression. Large-scale analysis of gene expression has the potential to generate large amounts of data, which should be made readily accessible to the scientific community. The Internet offers many potential advantages as a medium for the maintenance of these large data sets. CLINICAL RELEVANCE: Identification of structural, apoptosis-related, and disease-causing genes within the cornea by microarrays may increase the understanding of normal and abnormal corneal function with likely relevance to corneal diseases and transplants.
OBJECTIVES: To use microarray analysis to identify genes expressed in humandonor corneas and to create a preliminary, comprehensive database of humancorneal gene expression. METHODS: A complementary DNA (cDNA) library was constructed from transplant-quality, humandonor corneas. Biotin-labeled RNA was transcribed from the cDNA library and hybridized in duplicate to microarrays containing approximately 5600 human genes. Results were analyzed using a gene database of the National Institutes of Health, Bethesda, Md. Reverse transcriptase polymerase chain reaction analysis was performed to confirm corneal expression of genes identified by microarray analysis. RESULTS: Duplicate microarrays identified the expression of 1200 genes in humandonor corneas. Chromosomal loci had been assigned to 1025 (85%) of these genes. A preliminary database of humancorneal gene expression was compiled. A Web site containing these genes was created. Six collagen genes were identified that had not previously been localized within the cornea. Five apoptosis-related genes were identified, 4 of which had not previously been localized within the cornea. Three genes previously shown to cause corneal diseases were identified. Reverse transcriptase polymerase chain reaction analysis of genes identified by microarray analysis confirmed the corneal expression of 2 apoptosis-related genes and 1 collagen gene. CONCLUSIONS: Microarray analysis of healthy humandonor corneas has produced a preliminary, comprehensive database of corneal gene expression. Large-scale analysis of gene expression has the potential to generate large amounts of data, which should be made readily accessible to the scientific community. The Internet offers many potential advantages as a medium for the maintenance of these large data sets. CLINICAL RELEVANCE: Identification of structural, apoptosis-related, and disease-causing genes within the cornea by microarrays may increase the understanding of normal and abnormal corneal function with likely relevance to corneal diseases and transplants.
Authors: Justyna A Karolak; Karolina Kulinska; Dorota M Nowak; Jose A Pitarque; Andrea Molinari; Malgorzata Rydzanicz; Bassem A Bejjani; Marzena Gajecka Journal: Mol Vis Date: 2011-03-30 Impact factor: 2.367
Authors: Eranga N Vithana; Chiea-Chuen Khor; Chunyan Qiao; Ningli Wang; Tin Aung; Monisha E Nongpiur; Ronnie George; Li-Jia Chen; Tan Do; Khaled Abu-Amero; Chor Kai Huang; Sancy Low; Liza-Sharmini A Tajudin; Shamira A Perera; Ching-Yu Cheng; Liang Xu; Hongyan Jia; Ching-Lin Ho; Kar Seng Sim; Ren-Yi Wu; Clement C Y Tham; Paul T K Chew; Daniel H Su; Francis T Oen; Sripriya Sarangapani; Nagaswamy Soumittra; Essam A Osman; Hon-Tym Wong; Guangxian Tang; Sujie Fan; Hailin Meng; Dao T L Huong; Hua Wang; Bo Feng; Mani Baskaran; Balekudaru Shantha; Vedam L Ramprasad; Govindasamy Kumaramanickavel; Sudha K Iyengar; Alicia C How; Kelvin Y Lee; Theru A Sivakumaran; Victor H K Yong; Serena M L Ting; Yang Li; Ya-Xing Wang; Wan-Ting Tay; Xueling Sim; Raghavan Lavanya; Belinda K Cornes; Ying-Feng Zheng; Tina T Wong; Seng-Chee Loon; Vernon K Y Yong; Naushin Waseem; Azhany Yaakub; Kee-Seng Chia; R Rand Allingham; Michael A Hauser; Dennis S C Lam; Martin L Hibberd; Shomi S Bhattacharya; Mingzhi Zhang; Yik Ying Teo; Donald T Tan; Jost B Jonas; E-Shyong Tai; Seang-Mei Saw; Do Nhu Hon; Saleh A Al-Obeidan; Jianjun Liu; Tran Nguyen Bich Chau; Cameron P Simmons; Jin-Xin Bei; Yi-Xin Zeng; Paul J Foster; Lingam Vijaya; Tien-Yin Wong; Chi-Pui Pang Journal: Nat Genet Date: 2012-08-26 Impact factor: 38.330