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Quantitative analysis of acetylcholine in mouse brain tissue sections by Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences, MMS, Medical Mass Spectrometry.
2014 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Intro             This study revolves around the process of aging, with respect to acetylcholine production, which declines with increasing age and can cause dementia-related diseases. The acetylcholinesterase inhibitor tacrine was previously used in the treatment of Alzheimer’s disease and was used to study its effect with respect to increased acetylcholine production in young (12 weeks) and old (14 months) mice.

Method         Duplicates of frozen brain tissue were cut into sagittal and coronal sections with a thickness of 14 µm, and mounted on conductive indium tin oxide (ITO) glass slides. A total of four animals were used, one young tacrine treated, one old tacrine treated, one young control and one old control. Imaging and quantitation acetylcholine was performed using Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging (MALDI MSI).

Results, Discussion and Conclusion     This study presents a proof-of-principle of the quantitation of acetylcholine using MALDI MS Imaging. The results showed a 3-fold increase in acetylcholine in young mice, and a 2-fold increase in the old mice when comparing tacrine treated and control. The areas of higher concentrations correspond to the presynaptic neurons at the termination of the cholinergic pathways, which include the cortex, hippocampus, thalamus, hypothalamus and hindbrain. One exception is the cerebellum, which exhibited low concentrations in all tissues. The distribution of acetylcholine in the control mice was more evenly distributed. A tacrine dependent increase of acetylcholine in the basolateral amygdala was detected in both young and old mice, an area involved in fear-related learning. Another nucleus called the interpeduncular nucleus (IPN) exhibited a 3-4 fold increase compared to the rest of the midbrain. IPN was unaffected by the tacrin treatment and observed in all animals. This is explained by the dense cholinergic innervation from the medial habenula, which is an important link between forebrain and midbrain structures. 

Place, publisher, year, edition, pages
2014. , 31 p.
Keyword [sv]
Kvantifiering, avbildande masspektrometri, acetylkolin, tacrin, möss
National Category
Pharmacology and Toxicology
URN: urn:nbn:se:uu:diva-216794OAI: oai:DiVA.org:uu-216794DiVA: diva2:690823
Subject / course
Pharmaceutical Pharmacology
Educational program
Master of Science Programme in Pharmacy
Available from: 2014-05-28 Created: 2014-01-24 Last updated: 2014-05-28Bibliographically approved

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