Dissecting the molecular mechanisms of drosophila border cell migration using time-lapse live cell imaging

Dodgson, Lauren Dissecting the molecular mechanisms of drosophila border cell migration using time-lapse live cell imaging. Doctor of Philosophy thesis, University of Liverpool.

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Abstract

Dissection of the cellular dynamics and molecular pathways that drive collective cell migration is necessary to better understand cellular rearrangements that underpin normal development, as well as disease states such as cancer metastasis. Border cell migration in the Drosophila ovary has proven to be a good model of invasive cell migration, because of its genetic tractability, and also because recent advances in culturing egg chambers ex vivo have facilitated live cell imaging in this system. The aim of this thesis was to further develop and implement live cell imaging approaches, and to apply these to characterise the role of Pico, the Drosophila Mig10/RIAM/Lpd (MRL) protein in border cell migration. MRL proteins are known to regulate actin dynamics, but their role in epithelial cell migration had not been established. Through careful optimisation, suitable approaches were developed for: medium preparation; dissection and mounting of egg chambers; acquisition of images by confocal microscopy. A fluorescently-labelled reporter strain with improved optical properties was generated to monitor actin dynamics, and a number of other reporters were characterised, either alone or in combination, to determine their behaviour and effect on migration. After trialling several analytical tools and quantitative methods, a streamlined approach to analysing the image data was developed allowing: tracking of border cell migration in four dimensions (XYZ and time) to obtain information about behaviour of the migratory cells; measurement of cellular protrusion dynamics to obtain mechanistic insight into why cellular dynamics might change in different genetic backgrounds. Finally, these approaches were applied to the characterisation of Pico and its interacting partner SCAR, demonstrating that pico affects border cell migration through the modulation of actin protrusion dynamics in a SCAR-dependent manner.

Item Type:	Thesis (Doctor of Philosophy)
Additional Information:	Published article in Appendix Open Access Date: 2013-10 (completed)
Uncontrolled Keywords:	Drosophila, Border cell, migration, pico, MRL, SRF, Live-cell Imaging
Subjects:	?? QH301 ??
Divisions:	Faculty of Health and Life Sciences
Depositing User:	Symplectic Admin
Date Deposited:	06 Aug 2014 11:08
Last Modified:	17 Dec 2022 01:41
DOI:	10.17638/00016293
Supervisors:	Bennett, Daimark
URI:	https://livrepository.liverpool.ac.uk/id/eprint/16293