Supervisors

Introduction:

Alzheimer's disease (AD) is a progressive neurodegenerative disorder with no treatment. Overproduction of amyloid-β (Aβ) and subsequent deposits of Aβ plaques in brain tissue is the principal contributor to AD's pathogenesis. The blood-cerebrospinal fluid barrier plays a vital role in the clearance of Aβ from the brain and can inhibit Aβ aggregation and toxicity (Solár et al., 2020). On the other hand, neuroinflammatory responses associated with changes at the blood-CSF barrier have been directly implicated in the Aβ plaques deposit in the brain, as inflammatory cytokines released locally have been shown to both alleviate and aggravate the cluster formation (Brkic et al., 2015; Hu et al., 2009). However, the exact relationship is unclear. 

During the previous collaboration with JCMM, we learned that Z310 cells, an in vitro model of the blood-CSF barrier, are able to uptake the Aβ from the culture medium and subsequently respond to Aβ aggregation.

The current proposal aims to have a deeper look at signalling pathways involved in inflammatory reactions caused by Aβ aggregation in Z310 cells. We will use different techniques, including cell culture, immunocytochemistry (ICC), and imaging.

Supervisor: Alemeh Zamani, PhD (English-speaking)

 

Experimental Plan:

Seed cells in 24-well plates

(1)    Incubate the cells with 5µM Aβ1–42 for a varying period

(2)    Examine the viability of cells using live/dead assay

(3)    Check the level of different immune receptors using ICC

(4)    Check the level of different markers of the barrier integrity using ICC

Seed cells on Transwell plate

(1)    Apply the 5µM Aβ1–42 after barrier formation

(2)    Measure trans-epithelial voltage for barrier integrity alteration study

 

References:

Brkic, M., Balusu, S., Van Wonterghem, E., Gorlé, N., Benilova, I., Kremer, A., Van Hove, I., Moons, L., De Strooper, B., Kanazir, S., Libert, C., Vandenbroucke, RE, 2015. Amyloid β Oligomers Disrupt Blood-CSF Barrier Integrity by Activating Matrix Metalloproteinases. J. Neurosci. 35, 12766–12778. https://doi.org/10.1523/JNEUROSCI.0006-15.2015

Hu, X., Crick, S.L., Bu, G., Frieden, C., Pappu, R.V., Lee, J.-M., 2009. Amyloid seeds formed by cellular uptake, concentration, and aggregation of the amyloid-beta peptide. Proc Natl Acad Sci U S A 106, 20324–20329. https://doi.org/10.1073/pnas.0911281106

Solár, P., Zamani, A., Kubíčková, L., Dubový, P., Joukal, M., 2020. Choroid plexus and the blood–cerebrospinal fluid barrier in disease. Fluids Barriers CNS 17, 35. https://doi.org/10.1186/s12987-020-00196-2