Disruptive effect of oxidative stress on intracellular Ca2+ signaling
The intracellular Ca2+ signaling system is intricate and of utmost importance for the health of each individual cell and thus the organism as a whole. It may be attacked and disturbed by many or most pathophysiological conditions. Flaws in this system will lead to malfunctioning and sometimes cell death.
There is still much to learn about this system, and differences between cell types add to the complexity. In this project, we want to focus on the Ca2+ signaling system that controls salivary fluid secretion, and how this fine-tuned system is brought out of balance by increased oxidative stress, due to for instance radiation in cancer treatment or inflammation in the autoimmune disease Sjögren’s syndrome, or simply during normal aging. In all cases, reduced Ca2+ signaling via the SOCE mechanism (store-operated Ca2+
Figure 1. Ca2+ signaling in salivary gland acinar cells may be interrupted by several factors, and by several intracellular pathways. (Ambudkar et al., BBA Molecular Cell Research 1865 (2018) 1771-1777).
Your part of the project
- Intracellular Ca2+-signaling: Use the calcium-sensitive intracellular dye Fura-2 on salivary gland cell lines. Compare SOCE current before and after oxidative stress, use pharmacological blockers of the relevant signaling components such as STIM1 (an endoplasmic reticulum Ca2+ sensor), Orai1 (a cell membrane Ca2+ pore controlled by STIM1), TRPM2 (a Ca2+ channel that is redox-sensitive) and aquaporin 5 (a water channel) to assess the role of each protein in the observed stress-induced changes.
- Real time qPCR on cell lines: compare expression levels of the relevant signaling components before and after oxidative stress.
- Western blot on cell lines: compare protein expression levels of the same signaling components.
- If time: Fura-2 experiments and/or expression studies of the most affected protein(s) from part 1, 2 and 3 on salivary gland biopsies from Sjögren´s patients.
Supervisors / research group
Profs Hilde K. Galtung and Trude M. Haug at the Institute of Oral Biology will be main supervisors. The master student will join our research group consisting of several PhD students and technicians, ensuring all the help and support needed to perform the suggested experiments. Professor Marianne Fyhn will be the internal supervisor at IBV.
For more information, please contact us: