PNPLA1 Geninde Mutasyon Saptanan Konjenital Iktiyoz Vakalarında Yağ Damlacıklarının Lipofaji Aracılı Regülasyonunun Incelenmesi
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Patatin Like Phospholipase Domain Containing Protein-1 (PNPLA1) gene mutations are shown to be linked with Autosomal Recessive Congenital Ichthyoses (ARCI). PNPLA1 protein located on phospholipid membrane structure of lipid droplets plays crucial roles in biosynthesis and regulation of phospholipids. Investigating the effects of PNPLA1 gene mutations on the regulation of lipid droplets will help to enlighten the pathways that cause the pathology of this disease. For this purpose, in the scope of this thesis firstly the accumulation of lipid droplets were compared between the primer fibroblast cultures of patient and control individuals. According to the results, an abnormal increase in number and size of lipid droplets of patient fibroblast cells compared to control cells was detected. In addition, it was determined that PNPLA1 gene mutations have no effect on the cytoplasmic localization of the protein. In order to investigate lipophagy mediated regulation, colocalization of lipid droplets with designated autophagic markers (Atg5, LC3, LAMP1, Lysotracker® Red-DND-99) was studied using fluorescence staining methods. Discovery of colocalization of lipid droplets with autophagic markers is an indicator of degradation of lipid droplets with autophagy at basal levels. However, there was no colocalization of lipid droplets with autophagic markers was detected for patient fibroblast cells. As a result, finding of abnormal lipid accumulations and defects in autophagy mechanisms of patients fibroblast cells is an indicator of crucial role that PNPLA1 protein playing in lipid droplet regulation. Advanced functional analysis of this protein will definitely help us to shed light on the prevailing mechanisms of the pathology of this disease.