High-resolution morphometric analysis of synaptic vesicle formation

by Kristina Jevdokimenko
Doctoral thesis
Date of Examination:2024-07-05
Date of issue:2025-06-23
Advisor:Dr. Eugenio F. Fornasiero
Referee:Dr. Dr. Oliver Schlüter
Referee:Prof. Dr. Stefan Jakobs
Persistent Address: http://dx.doi.org/10.53846/goediss-11337

 

 

Files in this item

Name:20240528_PhD_Thesis_merged.pdf
Size:22.9Mb
Format:PDF

This file will be freely accessible after 2026-07-04.

Abstract

English

Synaptic vesicle (SV) biogenesis plays an important role in neurotransmission, yet the intricacies of SV formation remain a significant scientific challenge. In this PhD thesis, I seek to address both technical and biological challenges associated with studying SV biogenesis through three main objectives: First, I aimed to establish a multiplexing method that enables high-resolution visualization of multiple targets within the same sample. This objective required the development of advanced imaging techniques that can simultaneously resolve various SV components, thereby providing a comprehensive view of the molecular composition within neurons. Second, I collected a high-resolution stimulated emission depletion (STED) microscopy dataset focused on several key SV proteins in developing neurons. The use of STED microscopy was critical here, as it surpasses the resolution limitations of traditional light microscopy and allows for the visualization of SV proteins at a nanoscale level within the complex environment of developing hippocampal neurons. Third, I analyzed the distribution and nanocluster formations of different SV proteins across various organelles in the secretory pathway. By examining specific protein localization and clustering, I aimed to uncover the molecular mechanisms and spatial organization involved in SV biogenesis. Together, these integrated approaches are designed to overcome existing limitations in SV research, providing unprecedented insights into the assembly and organization of synaptic vesicles. The high-resolution model developed from this research offers a novel perspective on SV formation, enhancing our understanding of synaptic function and potentially informing therapeutic strategies for neurodevelopmental disorders. This work, therefore, makes a critical contribution to the field of neurobiology by advancing both methodological capabilities and biological knowledge of synaptic vesicle biogenesis in primary hippocampal neurons.
Keywords: synaptic vesicle; neurodevelopment; STED; SUM-PAINT