The osteoclast ruffled border is a highly convoluted, complex membrane that is necessary for bone resorption. It is thought to form following mass lysosomal fusion with the boneapposing plasma membrane and vesicular trafficking is vital for its formation and function. The aim of this PhD was to better understand the ultrastructure, formation and function of the ruffled border using TEM and advanced imaging techniques. Ruffled border reformation following calcitonin treatment was visualised and the stages of ruffled border formation were described. Ruffled borders in healthy and osteopetrotic osteoclasts were also imaged by TEM and characterised using a morphological grading system. The key findings of this thesis are as follows: (1) vacuoles, not lysosomes, are the primary contributors of membrane to the ruffled border and the membrane projections of the ruffled border form passively as a consequence of channel formation, not actively by membrane folding, (2) extracellular vesicles are located, and appear to be released, at the ruffled border. Various functional aspects of the ruffled border were also investigated. Vesicles near the ruffled border were identified and characterised by immunoelectron microscopy based on their content and morphology. We found no morphological defects in ruffled borders in mice deficient in Plekhm1. In osteoclasts derived from patients with a SNX10 mutation, we found that while the cells retained the capacity to form well-developed ruffled borders, they did so less often than healthy control osteoclasts. Importantly, we observed that even in a population of healthy osteoclasts, ruffled border morphology is highly heterogeneous because they are at different stages in the resorption cycle. In conclusion, the data in this thesis provide novel findings, previously unseen details regarding how resorbing osteoclasts interact with the bone surface, and have revealed unique insights into ruffled border morphology, formation and the vesicles with which it interacts.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:742395 |
| Date | January 2018 |
| Creators | McDermott, Emma |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=236980 |
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