ABeta mediated diminution of MTT reduction-an artefact of single cell culture?

• Autorzy: Ronicke R. , Klemm A. , Meinhardt J. , Schroder U.H. , Fandrich M. , Reymann K.G.
• Języki publikacji: EN

Abstrakty

EN

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT) reduction assay is a frequently used and easily reproducible method to measure beta-amyloid (Aβ) toxicity in different types of single cell culture. To our knowledge, the infl uence of Aβ on MTT reduction has never been tested in more complex tissue. Initially, we reproduced the disturbed MTT reduction in neuron and astroglia primary cell cultures from rats as well as in the BV2 microglia cell line, utilizing four different Aβ species, namely freshly dissolved Aβ (25-35), fi brillar Aβ (1-40), oligomeric Aβ (1-42) and oligomeric Aβ (1-40). In contrast to the fi ndings in single cell cultures, none of these Aβ species altered MTT reduction in rat organotypic hippocampal slice cultures (OHC). Moreover, application of Aβ to acutely isolated hippocampal slices from adult rats and in vivo intracerebroventricular injection of Aβ also did not infl uence the MTT reduction in the respective tissue. Failure of Aβ penetration into the tissue cannot explain the differences between single cells and the more complex brain tissue. Thus electrophysiological investigations disclosed an impairment of long-term potentiation (LTP) in the CA1 region of hippocampal slices from rat by application of oligomeric Aβ (1-40), but not by freshly dissolved Aβ (25-35) or fi brillar Aβ (1-40). In conclusion, the experiments revealed a glaring discrepancy between single cell cultures and complex brain tissue regarding the effect of different Aβ species on MTT reduction. Particularly, the differential effect of oligomeric versus other Aβ forms on LTP was not refl ected in the MTT reduction assay. This may indicate that the Aβ oligomer effect on synaptic function refl ected by LTP impairment precedes changes in formazane formation rate or that cells embedded in a more natural environment in the tissue are less susceptible to damage by Aβ, raising cautions against the consideration of single cell MTT reduction activity as a reliable assay in Alzheimerís drug discovery studies.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2009
• Tom: 69
• Numer: 1
• Opis fizyczny: p.94-95

Twórcy

autor

Ronicke R.

• Leibniz Institute for Neurobiology, Magdeburg, Germany

autor

Klemm A.

• Leibniz Institute for Neurobiology, Magdeburg, Germany

autor

Meinhardt J.

• Leibniz Institute for Neurobiology, Magdeburg, Germany

autor

Schroder U.H.

• Leibniz Institute for Neurobiology, Magdeburg, Germany

autor

Fandrich M.

• Leibniz Institute for Neurobiology, Magdeburg, Germany

autor

Reymann K.G.

• Leibniz Institute for Neurobiology, Magdeburg, Germany