Survey of Large Dense Core Vesicles (LDCVs) markers for better detection of compound exocytosis

Nagle, Prajwal (2023) Survey of Large Dense Core Vesicles (LDCVs) markers for better detection of compound exocytosis. Masters thesis, Indian Institute of Science Education and Research, Kolkata.

Text (MS dissertation of Prajwal Nagle (18MS115)) Thesis_18MS115.pdf - Submitted Version Restricted to Repository staff only Download (2MB)

Abstract

Compound exocytosis represents a specialized form of secretion in which vesicles undergo homotypic fusion either before (multi-vesicular exocytosis) or after fusion (sequential exocytosis) with the plasma membrane. Although numerous probes exist for studying simple exocytosis, it is challenging to identify compound exocytosis with them. To advance our understanding, we need a fluorescent probe that could detect SVs undergoing compound exocytosis and allow us to distinguish it from other modes of exocytosis. Here, we used two cargo proteins (NPY and tPA) labeled with different fluorescent proteins (mCherry, Venus, and eGFP) and employed total internal reflection fluorescence microscopy (TIRF-M) to capture distinct single-granule and multi-granular fusion events. We identified tPA-eGFP as a better probe for studying compound exocytosis, as it can reliably detect both sequential and multi-vesicular exocytosis. We have also successfully detected compound exocytosis in human β-cell, but its role still remains unclear. Further studies with the identified marker will help unravel the specific details and improve our understanding of compound exocytosis.

Actions (login required)

View Item