Involvement of the blood-cerebrospinal fluid barrier in Alzheimer's disease progression- an in-vitro study

Téma již má řešitele.
Řešitel
Anna Brodecká - Gymnázium Brno, Slovanské náměstí, příspěvková organizace
Instituce
Masarykova univerzita
Fakulta/ústav
Fakulta lékařská
Další údaje o pracovišti
Department of Anatomy
Lektoři
Alemeh Zamani
Podpora
JCMM podpořila toto SOČ téma částkou 20 Kč na materiál a částkou 10 000 Kč na honorář školitele.

 

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 both alleviate and aggravate the cluster formation (Brkic et al., 2015; Hu et al., 2009). However, the exact relationship is unclear. 

The culture system enables molecular manipulations to investigate the molecular mechanism underlying the blood-CSF barrier dysfunction in relation to AD.

Experimental Plan:

Z310 cell and HEK293 cell culture (control cells)

  1. Seed cells in 8-well chamber slides 

  2. Incubate the cells with 1µM FITC– Aβ1–42 

  3. Examine cellular uptake after 24h using microscopy 

  4. Incubate cells with Aβ1–42 for varying periods of time 

  5. Examine the viability of cells using MTT assay 

  6. Find the time required for HMW formation of Aβ1-42 using WB 

  7. Check cytokine release using a Multiplex array 

  8. Check TTR and TJs expression for barrier integrity using ICC 

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