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Salvador Pastor, Irene Rodriguez-Hernandez, Maria Jimena Rojas, Rogelio Gonzalez-Sarmiento, J.C Pastor; Lymphotoxin Alpha (lta) Expression In Human Normal Retina Retina 4 Project:(genetics Of Inflammation And Repairing Process In The Retina). Invest. Ophthalmol. Vis. Sci. 2011;52(14):5337.
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To investigate the possible functional role of a LTA polymorphism (SNP) rs2229094 (T-->C), previously described and associated to PVR development in a two phases study. Phase I consisted of to examine whether the mRNA of LTA is expressed in human retinas and peripheral blood in normal conditions. In phase II, currently ongoing, a vector will be create with the variant alleles and cell lines will be transfect. We communicate the results of the mRNA of LTA in normal human retinas and normal human blood samples
Total RNA was isolated from 3 whole human retinas obtained from fresh eyeballs (Regional tissue-Bank) and 2 peripheral blood samples from normal donors (after informed consent), using Trizol (Invitrogen). cDNA synthesis was performed with the IMPROM-II kit Promega from total RNA. RT-PCR was performed using flanking primers of cDNA and GAPDH was used as an internal control to ensure cDNA quality and loading accuracy. PCR products were separated by electrophoresis in TBE agarose gels and isolated from using a SephaGlass BandPrep kit (Amersham Biosciences ).The purified products were TA-cloned into pGEM-T Easy Vector System (Promega) and sequenced in an ABI373 DNA sequencer (Applied Biosystems).
Expression signals of LTA in all retina samples were below the significance level of the method. LTA gene was isolated in peripheral blood samples. Furthermore alternative transcriptions were obtained.
Results confirm that LTA does not show significant levels in normal retinas. Nevertheless retinas obtained from altered eyes, with RD and/or PVR could express higher values related to the blood-retinal barrier breakdown as suggested. These findings warrant further studies.
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