Investigation of putative signal transduction processes of normal prion protein and their role in spongiform encephalopathy genesis

• Forschungsstelle: EU FRP
• Projektnummer: 98.0162
• Projekttitel Englisch: Investigation of putative signal transduction processes of normal prion protein and their role in spongiform encephalopathy genesis

Schlüsselwörter (Englisch)

Prion protein; spongiform encephalopathy

Alternative Projektnummern (Englisch)

EU project number: BMH4CT98-6006

Forschungsprogramme (Englisch)

EU-programme: 4. Frame Research Programme - 4.2 Agriculture and agroindustry

Kurzbeschreibung (Englisch)

See abstract

Partner und Internationale Organisationen (Englisch)

Coordinator: Institute of Psychiatry, Dep. of Neuroscience (UK)

Abstract (Englisch)

During the funding period, the lab at the Institute of Neuropathology has been attempting to establish long term slice cultures from PrP over-expressing mice, PrP-null mice and wild-type mice for expression on PrP in adenoviral vectors. In collaboration with Beat Gaehwiler the Zurich lab has started to establish the organotypic hippocampal slice culture (OHSC) technique in order to study prion replication in vitro. Due to the long duration of prion replication in vivo, we had to ascertain a slice survival time of up to 8 weeks in the case of tga20 transgenic mice (and much more for wild-type mice) needs to be achieved. C57BL/6 wild-type, prion gene (Prnp) knock-out and PrP overexpressing mice (tga20) were sacrificed (postnatal day 6) by decapitation. Thereafter, a frontal slice of the caudal cerebrum was aseptically removed and transversally sectioned at 350 µm thickness on a tissue chopper. Slices were then transferred onto culture plate inserts displaying 0.4 µm membrane pores. Some of the slice cultures were irradiated with 1000 Rad directly after preparation in order to diminish glial proliferation within the cultures. OHSCs were kept in culture for up to 8 weeks; medium was changed every 3 days. OHSCs of all experimental groups were taken at different time points for histological analysis. Immunohistochemical stainings included GFAP (glial fibrillary acidic protein) for detection of astrocytes, CD11b (Mac-1) for displaying microglial cells, and MAP-2 (microtubule-associated protein) as well as synaptophysin for detection of neuronal structures. It was shown that after 7 to 10 days about 90% of all slices died. Only very few OHSCs stayed alive after 8 weeks in vitro, although various changes in the culture conditions were performed (e.g. medium, preparation). In summary, this sub-program is progressing satisfactorily but is not yet at a stage for downstream analysis.

Datenbankreferenzen (Englisch)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 98.0162