Advanced search
1 file | 29.48 MB

Exploring the role of A-type lamins in cellular oxidative stress

Tom Sieprath (UGent)
(2017)
Author
Promoter
(UGent) and (UGent)
Organization
Abstract
Every cell contains the genetic information needed to create an entire organism. This blueprint is stored in the cell nucleus. The nucleus continuously regulates the accessibility of this information based on ever-changing intra- and extracellular stimuli. Therefore, proper functioning of the nucleus is crucial for cellular and organismal survival. The nuclear lamina, a perinuclear network composed of type V intermediate filaments called lamins, is emerging as key regulator in nuclear organization. It physically shapes the nucleus, influences gene expression and modulates cell differentiation. A recent addition to the expanding list of functions of the nuclear lamina is an apparent involvement in cellular redox homeostasis. Indeed, cells from patients suffering from various laminopathies display increased levels of intracellular reactive oxygen species (ROS) and often show a higher susceptibility towards induced ROS. The underlying pathways however, remain poorly understood. The goal of this PhD dissertation was to obtain a better insight in this novel putative pathogenic feature. Chapter 1 comprises a general introduction into lamin biology and the state of the art with respect to their involvement in redox biology, next to a guide into fluorescence microscopy of redox-related processes. In Chapter 2, a quantitative comparison and characterization is presented of various experimental perturbations to interfere with lamin A metabolism in primary fibroblast cells. Based on the results shown in this chapter, perturbations were selected to create the models that were used in the experiments in chapters 4 & 5. In chapter 3, the development and benchmarking of a novel high-content microscopy method for the simultaneous measurement of intracellular ROS levels and mitochondrial function is outlined, together with a complementary automated analysis pipeline. The application of the newly developed method from Chapter 3 on the selected models from Chapter 2 culminated in the discovery that distinct lamin variants induce divergent oxidative responses, eventually resulting in different cell fates (Chapter 4), and pointed to the involvement of perturbed protein degradation pathways as a causal factor for oxidative stress (Chapter 5).

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 29.48 MB

Citation

Please use this url to cite or link to this publication:

Chicago
Sieprath, Tom. 2017. “Exploring the Role of A-type Lamins in Cellular Oxidative Stress”. Ghent, Belgium: Ghent University. Faculty of Bioscience Engineering.
APA
Sieprath, T. (2017). Exploring the role of A-type lamins in cellular oxidative stress. Ghent University. Faculty of Bioscience Engineering, Ghent, Belgium.
Vancouver
1.
Sieprath T. Exploring the role of A-type lamins in cellular oxidative stress. [Ghent, Belgium]: Ghent University. Faculty of Bioscience Engineering; 2017.
MLA
Sieprath, Tom. “Exploring the Role of A-type Lamins in Cellular Oxidative Stress.” 2017 : n. pag. Print.
@phdthesis{8534220,
  abstract     = {Every cell contains the genetic information needed to create an entire organism. This blueprint is stored in the cell nucleus. The nucleus continuously regulates the accessibility of this information based on ever-changing intra- and extracellular stimuli.
Therefore, proper functioning of the nucleus is crucial for cellular and organismal survival. The nuclear lamina, a perinuclear network composed of type V intermediate filaments called lamins, is emerging as key regulator in nuclear organization. It physically shapes the nucleus, influences gene expression and modulates cell differentiation. A recent addition to the expanding list of functions of the nuclear lamina is an apparent involvement in cellular redox homeostasis. Indeed, cells from patients suffering from various laminopathies display increased levels of intracellular reactive oxygen species (ROS) and often show a higher susceptibility towards induced ROS. The underlying pathways however, remain poorly understood. The goal of this PhD dissertation was to obtain a better insight in this novel putative pathogenic feature.
Chapter 1 comprises a general introduction into lamin biology and the state of the art with respect to their involvement in redox biology, next to a guide into fluorescence microscopy of redox-related processes. In Chapter 2, a quantitative comparison and characterization is presented of various experimental perturbations to interfere with lamin A metabolism in primary fibroblast cells. Based on the results shown in this chapter, perturbations were selected to create the models that were used in the experiments in chapters 4 \& 5. In chapter 3, the development and benchmarking of a novel high-content microscopy method for the simultaneous measurement of intracellular ROS levels and mitochondrial function is outlined, together with a complementary automated analysis pipeline. The application of the newly developed method from Chapter 3 on the selected models from Chapter 2 culminated in the discovery that distinct lamin variants induce divergent oxidative responses, eventually resulting in different cell fates (Chapter 4), and pointed to the involvement of perturbed protein degradation pathways as a causal factor for oxidative stress (Chapter 5).},
  author       = {Sieprath, Tom},
  isbn         = {9789463570435},
  language     = {eng},
  pages        = {VIII, XII, 140},
  publisher    = {Ghent University. Faculty of Bioscience Engineering},
  school       = {Ghent University},
  title        = {Exploring the role of A-type lamins in cellular oxidative stress},
  year         = {2017},
}