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Tuhina Prasad, Amrisha Verma, Qiuhong Li, ; Expression and Cellular Localization of Mas Receptor in the Adult and Developing Mouse Retina. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4470.
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Recent studies have provided evidence that a local rennin angiotensin system (RAS) exists in the retina and plays an important role in retinal neuro-vascular function. We have recently shown that increased expression of ACE2 and Ang1-7, two components of the protective axis of the RAS, in the retina via AAV mediated gene delivery, conferred protection against diabetes- induced retinopathy. We hypothesize that the protective molecular and cellular mechanisms of Ang1-7 are mediated by its receptor, Mas, and its expression level and cellular localization dictate the response to Ang 1-7 and activation of subsequent protective signaling pathways. We tested this hypothesis by examining the expression and cellular localization of the Mas receptor in adult and developing mouse retina.
Eyes from adult and postnatal day 1(P1), P5, P7, P15 and P21 mice were enucleated and fixed in 4% paraformaldehyde overnight at 4°C.The cellular localization of Mas receptor protein was determined by immunofluorescence from OCT-embedded frozen sections or paraffin embedded sections using Mas receptor specific antibody, and the mRNA was detected by in situ hybridization from paraffin embedded sections using RNAscope kit from Advanced Cell Diagnostics Inc. Western blotting and real-time RT-PCR analysis were performed to determine the relative levels of Mas protein and mRNA in retina and cultured retinal cells.
In the adult eye, Mas receptor protein is abundantly present in the retinal ganglion cells (RGC) and the photoreceptor cells, lower level of expression is seen in endothelial cells, muller glial cells and other neurons in the Inner plexiform layer of the retina. In developing retina, both Mas receptor mRNA and protein expression can be detected in the inner retina at P1 and the expression levels increase with age to reach the adult level by P21.
Mas receptor is expressed in both adult and developing mouse retina, more abundant in retinal neurons than endothelial and muller glial cells. These observations suggest that Mas receptor mediated signaling may play important roles that extend beyond the well-established vascular effects of Ang 1-7.
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