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DM La Morticella, JR Samples, KC Rust, TS Acott, MK Wirtz; A Comparison of Porcine Ocular Tissue Gene Expression by Microarray Analysis . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2436.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Microarray analysis was used to determine if porcine cDNA would hybridize cleanly to human cDNA arrays. Gene expression from ocular tissues was measured and compared to help elucidate their functions. It was sought to determine if unique gene upregulation in different ocular tissues occurs, and if so what genes are involved. Methods: Total RNA was extracted from porcine iris, cilliary body, retina, and optic nerve tissues. Labeled cDNA was generated using the Perkin Elmer TSA kit. Samples were hybridized to human cDNA microarrays containing 5700 PCR products spotted in duplicate. Amplicons making up the arrays are genes or ESTs, and are amplified from ResGen human library clones. Labeled cilliary body, optic nerve, and retina cDNA were each hybridized separately with labeled iris cDNA on two double spotted arrays. Results: It was found that there are genes which are upregulated uniquely in each of the ocular tissues studied. Cilliary body uniquely upregulated genes included lysyl oxidase - like1 (LOXL1), ATP1 B3 ATPase, Na+/K+ transporting, beta 3 polypeptide, FMOD Fibromodulin, GPNMB Glycoprotein (transmembrane) nmb, and OAT Ornithine aminotrasferase (gyrate atrophy). In Iris Beta A1 Crystallin (CRYBA1), and Actin gamma 2 smooth muscle enteric (ACTG2) were uniquely upregulated. Myelin Basic Protein (MBP), Proteolipid protein 1 (PLP1), Myelin - associated oligodendrocyte basic protein (MOBP), and S100 calcium - binding protein beta (SB100) were unique to optic nerve. In Retinal tissue CKMT2 creatine kinase mitochondrial 2 (sarcomeric), UNC119 (c. elegans) homolog, and ATPB2 ATPase Na+/K+ transporting beta 2 polypeptide were found to be uniquely upregulated. Conclusion: Upregulated genes in porcine ocular tissues could be determined by microarray analysis with SMChum5700 human cDNA arrays from the Spotted Microarray Core at Oregon Health and Science University. Genes that are upregulated specific to tissue type are able to be determined using these arrays.
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