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A Plate-Based Assay for the Measurement of Endogenous Monoamine Release in Acute Brain Slices
Published on: August 11, 2021
A novel method for the rapid determination of beta-amyloid toxicity on acute hippocampal slices using MTT and LDH
Emese Mozes1, Akos Hunya, Aniko Posa
1Department of Medical Chemistry, University of Szeged, Szeged, Hungary. mese859@gmail.com
Abstract:
It is difficult task to measure precisely the toxic effect of beta-amyloid (Aβ 1-42) peptides and also the protective effect of novel drug candidates against Aβ-peptides. The widely used MTT-assay in cell lines or primary cell cultures could be insensitive against Aβ-peptides. We describe here an easy and relevant method for testing Aβ 1-42 toxicity on acute hippocampal slices derived from rat. Brain slice viability in different conditions was measured using MTT and LDH assays. The concomitant use of these two assays can give detailed and relevant results on the toxic effect of Aβ 1-42 in oxygen-glucose deprived (OGD) acute brain slice model. Both assays are capable of quantifying tissue viability by measuring optical density (OD). We found that simultaneous application of OGD and Aβ 1-42 treatment induced a more intensive decrease in hippocampal slice viability than their separate effects. The use of MTT and LDH assay for quantifying brain slice viability proved to be an easy ex vivo method for investigating Aβ toxicity. Testing brain slices is more relevant in Alzheimer's Disease research than using in vitro cell cultures, due to maintenance of the three dimensional cellular network, the cell variability and intact cell connections.
Insights
Measuring beta-amyloid (Aβ 1-42) toxicity is challenging. This study presents an easy ex vivo method using MTT and LDH assays on rat brain slices, offering a relevant approach for Alzheimer's research.
Area of Science:
- Neuroscience
- Toxicology
- Biochemistry
Background:
- Assessing beta-amyloid (Aβ) peptide toxicity and drug efficacy is difficult.
- Standard MTT assays in cell cultures may lack sensitivity to Aβ toxicity.
Purpose of the Study:
- To introduce a straightforward and relevant ex vivo method for evaluating Aβ 1-42 toxicity.
- To compare the sensitivity of MTT and LDH assays in an acute brain slice model.
Main Methods:
- Utilized acute hippocampal slices from rats.
- Measured brain slice viability using MTT and Lactate Dehydrogenase (LDH) assays.
- Applied oxygen-glucose deprivation (OGD) combined with Aβ 1-42 treatment.
Main Results:
- Simultaneous OGD and Aβ 1-42 treatment significantly reduced hippocampal slice viability.
- The combined use of MTT and LDH assays provided detailed and relevant toxicity data.
- This ex vivo method proved effective in quantifying Aβ toxicity.
Conclusions:
- The MTT and LDH assay combination offers an easy ex vivo method for Aβ toxicity assessment.
- Brain slice models are more relevant than cell cultures for Alzheimer's research due to preserved cellular architecture.
- This method aids in investigating Aβ toxicity and potential therapeutic interventions.
