scholarly journals Targeted UHPLC-ESI-MS/MS Analysis of Selected Neurotransmitters, Tryptophan and Its Metabolite Kynurenine in Tau Transgenic Rat Brain Tissue: A Pivotal Study

Separations ◽  
2022 ◽  
Vol 9 (1) ◽  
pp. 16
Author(s):  
Juraj Piestansky ◽  
Andrea Forgacsova ◽  
Dominika Olesova ◽  
Jaroslav Galba ◽  
Peter Mikus ◽  
...  
Keyword(s):  
Brain Tissue ◽  
Aspartic Acid ◽  
High Performance ◽  
Aminobutyric Acid ◽  
Transgenic Rat ◽  
Elevated Concentration ◽  
Tissue Samples ◽  
Validation Parameters ◽  
On Line ◽  
The Central Nervous System

Neurotransmitters (NT) are widely distributed in the central nervous system. These molecules are important for many physiological processes and the function of the immune system. Imbalance of NT are linked to numerous neurological disorders and diseases, including tauopathies. Here, a targeted approach based on on-line combination of ultra-high performance liquid chromatography with tandem mass spectrometry was validated and applied to the quantitative analysis of nine NT (acetylcholine, choline, aspartic acid, asparagine, glutamic acid, glutamine, pyroglutamate, γ-aminobutyric acid, N-acetyl-L-aspartic acid), tryptophan and its metabolite kynurenine in brain tissue samples of a rat model for tauopathy. The applied analytical method was characterized by excellent validation parameters for all analytes, such as limits of detection in the range of 0.01–1.70 µg/mL, regression coefficients of the calibration curves ≥ 0.9946, intra-day and inter-day precision expressed as coefficient of variation in the range of 0.6–11.9% and 0.6–14.4%, and accuracy in the range of 87.6–107.1% and 87.2–119.6%. Our analytical approach led to the identification of increased levels of choline and γ-aminobutyric acid in pons, and elevated concentration levels of pyroglutamate in medulla oblongata. These findings indicate that NT could play a valuable role in the study and clarification of neuroinflammation and neurodegenerative diseases.

Visualisation of GABAergic neurons and synapses in the rat brain using immunohistochemistry for two forms of glutamate decarboxylase

2021 ◽  
Vol 21 (2) ◽  
pp. 63-73
Author(s):  
Valeria A. Razenkova ◽  
Dmitrii E. Korzhevskii
Keyword(s):  
Brain Tissue ◽  
Glutamate Decarboxylase ◽  
Brain Structures ◽  
Gabaergic Neurons ◽  
Laboratory Animals ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Background Staining ◽  
The Central Nervous System ◽  
Rat Forebrain

BACKGROUND: Taking into account the importance of GABAergic brain system research and also the opportunity to achieve specific and accurate results in laboratory studies using immunohistochemical approaches, it seems important to have a reliable method of visualization GABA-synthesizing cells, their projections and synapses, for the morphofunctional analysis of GABAergic system both in normal conditions and in the experimental pathology. AIM: The aim of the study was to visualize analyze GABAergic neurons and synapses within rats brain using three different antibody types against glutamate decarboxylase and to identify the optimal conditions for reaction performing. MATERIALS AND METHODS: The study was performed on paraffin brain tissue sections of 5 adult Wistar rats. Immunohistochemical reactions using three antibody types against glutamate decarboxylase isoform 67 (GAD67) and glutamate decarboxylase isoform 65 (GAD65) were performed. Additional controls on C57/Bl6 mice and Chinchilla rabbits brain samples were also carried out. RESULTS: Antibodies used in the research made it possible to achieve high quality of GABAergic structures visualizing without increasing background staining. At the same time different antibody types are distinct in their efficacy to perform immunohistochemistry reaction on laboratory animal brain tissue samples. By performing additional controls, we discovered that there is necessary to adsorb secondary reagents immunoglobulins in order to eliminate nonspecific staining. It was found that GAD67 and GAD65 distribution in rat forebrain structures is different. It was stated that GAD67 immunohistochemistry most completely reveals GABAergic brain structures compared to GAD65 immunhistochemistry. The possibility of determining morphological features of GABAergic neurons and synaptic terminals, as well as performing quantitative analysis, was demonstrated. CONCLUSIONS: The approach proposed makes it possible to specifically visualize GABAergic structures of the central nervous system of different laboratory animals. This could be useful both in fundamental studies and in pathology research.

scholarly journals A Simple Densitometric Method for the Quantification of Inhibitory Neurotransmitter Gamma-Aminobutyric Acid (GABA) in Rat Brain Tissue

2011 ◽  
Vol 2011 ◽  
pp. 1-5 ◽  
Author(s):  
Chidambaram Saravana Babu ◽  
Krishnamoorthy Selvarajan Kesavanarayanan ◽  
Periyathambi Kalaivani ◽  
Vijayan Ranju ◽  
Muthiah Ramanathan
Keyword(s):  
Rat Brain ◽  
Brain Tissue ◽  
High Performance ◽  
Aminobutyric Acid ◽  
Gamma Aminobutyric Acid ◽  
Inhibitory Neurotransmitter ◽  
Recovery Study ◽  
Layer Chromatography ◽  
Hptlc Method ◽  
Rat Brain Tissue

Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian central nervous system which is involved in various physiological and pathological processes. The present study demonstrates a simple high-performance thin-layer chromatography (HPTLC) method which was developed for the estimation of GABA in rat brain tissue. The method was validated in terms of precision, recovery, reproducibility, and variability. Instrumental precision was found to be 0.5891% CV and reproducibility of the method was found to be 0.4141% CV. Interday and intraday precision of the method was found to be 0.9453% and 1.3236% CV, respectively. Accuracy of the method was checked by the recovery study, and the average recovery of GABA was found to be 97.98% at 40 ng and 96.15% at 80 ng levels. The present HPTLC method for GABA estimation was found to be simple, precise, reproducible, sensitive, and accurate. No doubt, this proposed method will be a useful tool for the estimation of GABA in rat brain tissue.

scholarly journals Simultaneous HPLC–MS–MS quantification of 8-iso-PGF2α and 8,12-iso-iPF2α in CSF and brain tissue samples with on-line cleanup

2010 ◽  
Vol 878 (24) ◽  
pp. 2209-2216 ◽  
Author(s):  
Magdalena Korecka ◽  
Christopher M. Clark ◽  
Virginia M.-Y. Lee ◽  
John Q. Trojanowski ◽  
Leslie M. Shaw
Keyword(s):  
Brain Tissue ◽  
Tissue Samples ◽  
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