Senior Project Advisor
Project - Campus-only Access
centriolar satellites, Golgi apparatus, microscopy, immunofluorescence, image analysis, cell biology
Centriolar satellites are small, membraneless, proteinaceous granules that surround the centrosome and pericentriolar material, trafficking throughout the cell. They aid in centrosome maturation through the transport of centrosomal proteins and in the nucleation of the primary cilium. They have also been implicated in autophagy and proteostasis in the cell at large. While they organize at the centrosome, the factors influencing satellite location within the cell as a whole are unclear. Satellites are known to localize to noncentrosomal MTOCs in cell lines lacking the centrosome, indicating additional factors play a role in their localization. It is unknown whether secondary MTOCs influence satellite position. The Golgi is the cellular center of post-translational modification and a secondary MTOC, nucleating its own microtubules for protein export. To examine a possible linkage between the Golgi apparatus and centriolar satellites, hTERT-RPE1 cells were immunofluorescently stained for satellites, the Golgi, the centrosome, and imaged on Western Washington University’s Leica Stellaris 8 confocal microscope. Loss of organization during mitosis of centriolar satellites and the Golgi was observed to coincide at the prometaphase/metaphase transition. Satellite distribution was then quantified using custom ImageJ macros. Satellite intensity at the centrosome was found to be lower by at least 12% in cells where the centrosome was spatially separated from the Golgi, across five replicates. Centrosome separation from the Golgi also induced a quantifiable directional shift in satellite distribution in the direction of the Golgi in satellites both localized to the centrosome and in the cytoplasm. Taken together, the data suggest a role for the Golgi apparatus in organizing a secondary population of centriolar satellites.
(The document here is a placeholder for work to be published elsewhere, then linked to this page upon publication. A poster is also included in the file below.)
Goodfried, Jonah and Galati, Nick, "Assessing Positioning of Centriolar Satellites by the Golgi Apparatus" (2023). WWU Honors College Senior Projects. 721.
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