Skip to main content
Open Access Publications from the University of California

UC Irvine

UC Irvine Electronic Theses and Dissertations bannerUC Irvine

Spatiotemporal organization of cellular machinery underlying calcium signaling

  • Author(s): Ellefsen, Kyle Lee Kreitlein
  • Advisor(s): Parker, Ian
  • et al.
Creative Commons 'BY' version 4.0 license

The spatiotemporal diversity of Ca2+ signals enables them to mediate diverse functions inside the same cell; this diversity arises in part from the position, arrangement, and communication between Ca2+ permeable channels. The work presented in this thesis describes various approaches for determining how these complex Ca2+ signals are generated. The first part of this dissertation describes the construction of two types of microscopes which enhance spatiotemporal resolution of Ca2+ signals. The spinning-spot shadowless total internal reflection fluorescence microscopy (chapter 2) provides enhanced shadow-free imaging of Ca2+ signals arising near the plasma membrane. A simplified lattice light-sheet microscope (chapter 3) enables high spatial resolution and 3D visualization of Ca2+ signals throughout the cell. This dissertation also describes new analytic techniques for understanding local Ca2+ signals. Chapter 4 describes a new set of algorithms and software for detecting and analyzing of local calcium signals. Chapter 5 details the precision with which the location of channels underlying local Ca2+ signals can be determined. In the next two chapters, these techniques are applied to study signaling pathways involving two important Ca2+ permeable channels. In chapter 6, Ca2+ signals generated by the inositol trisphosphate receptor (IP3R) are used to infer the diffusion coefficient of its agonist, IP3, leading to the conclusion that IP3 is a local, rather than global, messenger. Chapter 7 details work which shows that the mechanosensitive Ca2+ channel Piezo1 diffuses in the plasma membrane and that its activity is enriched in regions where cells generate mechanical traction forces.

Main Content
For improved accessibility of PDF content, download the file to your device.
Current View