The development of microfluidic assays for functional neural network communication studies and CNS drug discovery

Student thesis: Doctoral Thesis

Abstract

The study of neuroscience and the research tools used to perform this research is fundamental to the understanding of the human brain under physiological and pathophysiological conditions. Indeed, the prevalence of CNS disorders in the global population has sharply increased over the last two decades, and new techniques are required to dissect the complexities of the underlying mechanisms.Whilst in vivo research presents the closest means of replicating human CNS disorders, there are significant translational challenges in replicating these diseases using animal models. Breaking down the complexities present within the brain to assess single cellular mechanisms sequentially may be facilitated by in vitro methodologies, which provide invaluable information on neural network function under controlled conditions.A surge in microfluidic technology over the last 15 years has enabled considerable advances in the development of new in vitro research tools for neuroscientific research, offering greater control over experimental conditions including neural network patterning and fluid handling. Microfluidic devices are often transparent and thus can be readily interfaced with microscopy for optical imaging assays following chemical stimulation of neuronal cultures.This thesis explores novel avenues for microfluidic assay development using dual chamber microfluidic devices containing environmentally isolated, but synaptically connected neural networks. First, voltage imaging assays are considered as an alternative to Ca2+ imaging assays to improve upon the temporal resolutions of standard optical imaging, whilst maintaining a higher data throughput when compared to electrophysiological whole cell patch clamp recordings. Then, the need for manual drug applications are resolved by means of developing a microfluidic perfusion system for early CNS drug discovery.Finally, chemogenetic assays are employed in combination with Ca2+ imaging and microfluidic perfusion to selectively stimulate a sub-population of transfected neurons whilst monitoring the subsequent cascade of activity in the surrounding neural network. In conclusion, the microfluidic assays developed can be used for studying neurophysiological mechanisms of synaptic communication, are capable of screening CNS acting drugs, and lay the groundwork for alternative methods to manipulate the activity of neural networks.
Date of Award30 Jul 2020
Original languageEnglish
Awarding Institution
  • University Of Strathclyde
SponsorsNational Centre for the Replacement, Refinement and Reduction of Animals in Research NC3Rs & University of Strathclyde
SupervisorMichele Zagnoni (Supervisor) & Trevor Bushell (Supervisor)

Cite this

'