Thesis
Exploiting surface functionalisation and micropatterning for live cell imaging of mitochondrial trafficking in hippocampal neural cultures
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2025
- Thesis identifier
- T17529
- Person Identifier (Local)
- 201653348
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Mitochondrial motility is essential for neuronal function, ensuring proper ATP distribution, calcium buffering, and synaptic maintenance. However, tracking mitochondrial transport in neurons remains challenging due to their highly branched and overlapping processes. The conditions used to grow neurons in vitro, including the substrate on which the cells are grown, impact morphology, branching, extent of neurite growth, and potentially intracellular transport, including mitochondrial transport and dynamics, yet the specific effects on mitochondrial motility remain unclear. This thesis investigates mitochondrial motility in primary hippocampal neurons cultured on traditionally used substrates - poly-D-lysine (PDL), laminin and dual PDL/laminin – and on chemically charged substrates: carboxyethylsilanetriol (CES) and aminopropyltriethoxysilane (APTES). Additionally, microcontact printing was employed to create micropatterned substrates to restrict neurite outgrowth and enable structured mitochondrial tracking. To optimise micropatterning for neuronal cultures, key parameters—including stamp wettability, bio-ink composition and substrate blocking— were adjusted to enhance pattern fidelity and reduce off-pattern adhesion. Live-cell imaging and kymograph analyses were used to quantify mitochondrial motility within neurons grown on different substrate conditions. Compared to traditional coatings, CES-coated substrates significantly reduced the fraction of motile mitochondria, segmental velocities, and run lengths, while increasing pause duration and frequency. APTES substrates supported moderately higher motility than CES, indicating that surface charge influences cytoskeletal interactions and organelle transport. Additionally, neuron-to-astrocyte ratios were higher on CES and APTES surfaces, suggesting substrate chemistry impacts cell composition. Micropatterned substrates induced further reductions in motile fractions and travel distances across all conditions, with a significant shift in mitochondrial pausing behaviour. These findings suggest that both biochemical and spatial constraints influence neurons to affect intracellular organelle transport, providing a controlled framework for studying neuronal trafficking mechanisms. This study establishes a reproducible platform for quantifying mitochondrial motility under defined spatial constraints and highlights how substrate composition and patterning can modulate intracellular transport. These insights offer a foundation for future studies investigating transport deficits in neurodegenerative diseases and axonal injury.
- Advisor / supervisor
- Sandison, Mairi
- Chalmers, Susan
- Resource Type
- DOI
Las relaciones
Elementos
| Miniatura | Título | Fecha de subida | Visibilidad | Acciones |
|---|---|---|---|---|
|
|
T17529.pdf | 2025-12-11 | Público | Descargar |