Differential regulation of caveolae and clathrin-coated pits by cell tension and its heterogeneity

Roy, Rohan (2025) Differential regulation of caveolae and clathrin-coated pits by cell tension and its heterogeneity. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Rohan Roy (20MS110)) 20MS110_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (5MB)

Abstract

Caveolae plays a vital role in mechanoprotection, signaling, endocytosis, and mechanosensing in cell mechanobiology. An omega-shaped domain in the plasma membrane, having a size range between 60 and 100 nm, caveolae are composed of Caveolin-1 proteins supported by cavin-1 trimers. Of many functions performed by Cav1, membrane tension buffering is the most wellknown, and its intricate details have been previously investigated in our lab. As a reservoir of extra membrane, Cav1 buffers tension change by flattening out with the release of Cavin-1 proteins leaving only Cav1-scaffolds or becoming curved with more membrane to form dolines. Due to this property being served through tension mediation, the presence of crosstalk between Cav1 and CME pathway, among others, is unsurprising. Cross-talk investigation between Cav1 and the CME pathway around the tension landscape was done by estimating clathrin density and endocytosis rate marked by transferrin cargo across the basal surface of the cell from TIRF movie for Cav1 RFP and Cav1 KO cell lines. Intensity based kymograph analysis revealed Cav1RFP cells have more number of CCP pit density than Cav1KO from the midsection towards the cell periphery showing Cav1 aids in CCP pit formation although tension rules. Further object based detection revealed 3 separate classes of Cav1 population being Caveolae, scaffold and internalized Cav1 and the cell center vs periphery analysis showed that majority of the scaffold population is present towards the periphery which might be a proof of concept of cross-talk and would require further investigation. 1

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