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Julie Thu A. Tran, Anisse Saadi, Abul K. Rahman, MD Nawajes A. Mandal; Developmental Expression, Tissue Distribution and Localization of CERKL in Murine Retina. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1845.
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© ARVO (1962-2015); The Authors (2016-present)
Ceramide kinase like (CERKL) is one of the newest members of Retinitis pigmentosa (RP) genes, mutations in which are associated with recessive, nonsyndromic retinitis pigmentosa (RP26) with significant macular involvement during the early stages of the disease. Although CERKL has homology with Ceramide kinase protein (CERK), the function of this protein is not yet known. The purpose of this study was to test the expression, distribution and localization of this gene and its protein products in mouse and rat tissues in order to better understand the role of this protein in complex human retinal pathology.
Expression and tissue distribution of Cerkl mRNA was determined by quantitative RT-PCR in adult mouse and rat tissue, in developing mouse eyes/retinas. Expression of the Cerk mRNA was also determined in all the tissues in order to understand the relationship of Cerkl with Cerk. Expression of the protein was determined by anti-CERKL antibody available from Abcam Inc. by Western blotting. Localization of the protein in mouse retina was determined by immunofluorescence microscopy.
Among all the rat tissue tested, relative mRNA expression of Cerkl was highest in the retina; >10-fold higher than the other Cerkl-expressing tissue such as brain and lungs. Cerk, on the other hand, showed higher expression in both brain and retina and the relative abundance of Cerk is >5 fold of Cerkl in the retina. There is only minimal level of expression of both the genes in developing eyes but the expression reaches to the peak at the maturity of the retina (2 months) and the level is maintained until late age (tested in 7 months in mouse and 14-18 months in rats). Presence of more than one CERKL protein bands was observed in the cytosolic fraction of the retina, but only one form in the nuclear fraction. In retinal section CERKL protein was localized prominently to the ganglion cells, inner nuclear layers, RPE cells; and sparsely in the photoreceptor inner segments.
We determined the expression of Cerkl mRNA in murine retina and the specific localization of the protein in different retinal cells. We hope this information will help in understanding the role of CERKL protein in retina and how mutations in this gene cause human retinal degeneration.
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