Poster #029, Childrens Hospital Los Angeles
The Cellular Uptake of Polystyrene Microplastics (PS-MPs) within Glomerular Endothelial Kidney Cells
Mentor: Mohammadreza Kasravi
Microplastics have emerged as a prevalent health issue. Their presence in everyday commodities: water bottles, chewing gum, toothbrushes, and more makes them a constant threat. More importantly, research has revealed that microplastics are significant drivers of cell death, mitochondrial dysfunction, oxidative stress, and inflammation; additionally, our lab has already shown that renal cells uptake microplastics. This project aims to assess the whereabouts and fate of microplastics inside Glomerular Endothelial Cells. GECs were treated with red fluorescent polystyrene microplastics (PS-MPS) (1µm), and subsequently fixed at 1hr, 3hr, 6hr, and 24hrs. GECs were stained with primary antibodies: EEA1 (Early Endosome marker), LAMP1 (Lysosome marker), Rab7 (Late Endosome Marker), Rab11 (Excretory Vesicle marker) and secondary antibodies; confocal fluorescence microscopy imaging was used to determine intracellular fate, or which microplastics were being absorbed by the endo/lysosomes of the cell. In particular, their presence within specific intracellular components was determined by assessing the overlap of microplastics (red markers) and endo/lysosomes (green markers). As expected, PS-MPS were readily uptaken by GECs. We confirmed that PS-MPS co-localize with intracellular structures at 24 hours. At the end of our analysis, we expect that the overlie of the red and green markers will reveal that a certain percentage of PS-MPS were recognized as substrates and uptaken via endocytosis, gathering in the perinuclear region of the cell, while another percent of cells exit the cell via exocytosis. Learning more about the intercellular fate of PS-MPS within renal kidney cells is essential to understand and alleviate the adverse effects of PS-MPS on kidney homeostasis.