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Study of the impact of Tau phosphorylation on its internalization and processing in cultured neuronal cells

Mathilde Wauters1*, Paula Paci1 and Laurence Ris1
  • 1 Umons, Faculty of Medicine and Pharmacy, Belgium

Several studies have already revealed the propagation of Tau protein aggregates in different brain regions according to a stereotypical scheme that might explained the successive onset of symptoms in Alzheimer’s disease. In comparison, the implication of soluble Tau proteins in this transmission is poorly reported in the literature even though it seems that they constitute the most toxic species. In this study, we hypothesized that the phosphorylation of the protein, which is implied in its intracellular aggregation, could also play a role in this transmission. We applied two recombinant monomeric Tau proteins presenting a distinct phosphorylation state in the extracellular space of neuroblastoma cells (N2A) and of primary embryonic cortical cells. Proteins were detected by immunocytochemistry through a his-Tag included at their C-terminal extremity. We observed that the phosphorylation increased protein internalization by up to 10 times within both cell types. However, cellular distribution and processing of the phosphorylated protein were different in N2A cells and in cortical neurons. In N2A cells, proteins were found mainly around the nucleus up to 3 days after the period of incubation. Conversely, in cortical cells, proteins were much more distributed in soma and neurites and disappeared 24h after the end of the incubation period. This could be explained either by a digestion or an exocytosis of the protein. These two hypotheses will be tested by further experiments and the formation of intra-cytoplasmic aggregates will be studied in the presence of phosphorylated or non-phosphorylated proteins. These results could help to better understand the role of extracellular tau in the propagation of the disease and in the associated synaptic transmission deficits.

Acknowledgements

We would like to thank Progenosis who generously supplied us with non-phosphorylated and phosphorylated Tau proteins

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Keywords: tau protein, Extracellular Space, Phosphorylation, neuronal cell culture, propagation

Conference: 11th National Congress of the Belgian Society for Neuroscience, Mons, Belgium, 22 May - 22 May, 2015.

Presentation Type: Poster presentation

Topic: Neuroscience

Citation: Wauters M, Paci P and Ris L (2015). Study of the impact of Tau phosphorylation on its internalization and processing in cultured neuronal cells. Front. Neurosci. Conference Abstract: 11th National Congress of the Belgian Society for Neuroscience. doi: 10.3389/conf.fnins.2015.89.00001

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Received: 05 May 2015; Published Online: 05 May 2015.

* Correspondence: Miss. Mathilde Wauters, Umons, Faculty of Medicine and Pharmacy, Mons, Hainaut, B-7000, Belgium, mathilde.wauters@student.umons.ac.be