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  • Label-free lipidome study of paraventricular thalamic nucleus (PVT) of rat brain with post-traumatic stress injury by Raman imaging.

Label-free lipidome study of paraventricular thalamic nucleus (PVT) of rat brain with post-traumatic stress injury by Raman imaging.

The Analyst (2020-11-03)
Ardalan Chaichi, Syed Mohammad Abid Hasan, Nishir Mehta, Fabrizio Donnarumma, Philip Ebenezer, Kermit K Murray, Joseph Francis, Manas Ranjan Gartia
ABSTRACT

Post-traumatic stress disorder (PTSD) is a widespread psychiatric injury that develops serious life-threatening symptoms like substance abuse, severe depression, cognitive impairments, and persistent anxiety. However, the mechanisms of post-traumatic stress injury in brain are poorly understood due to the lack of practical methods to reveal biochemical alterations in various brain regions affected by this type of injury. Here, we introduce a novel method that provides quantitative results from Raman maps in the paraventricular nucleus of the thalamus (PVT) region. By means of this approach, we have shown a lipidome comparison in PVT regions of control and PTSD rat brains. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was also employed for validation of the Raman results. Lipid alterations can reveal invaluable information regarding the PTSD mechanisms in affected regions of brain. We have showed that the concentration of cholesterol, cholesteryl palmitate, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, sphingomyelin, ganglioside, glyceryl tripalmitate and sulfatide changes in the PVT region of PTSD compared to control rats. A higher concentration of cholesterol suggests a higher level of corticosterone in the brain. Moreover, concentration changes of phospholipids and sphingolipids suggest the alteration of phospholipase A2 (PLA2) which is associated with inflammatory processes in the brain. Our results have broadened the understanding of biomolecular mechanisms for PTSD in the PVT region of the brain. This is the first report regarding the application of Raman spectroscopy for PTSD studies. This method has a wide spectrum of applications and can be applied to various other brain related disorders or other regions of the brain.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
1,2-Diacyl-sn-glycero-3-phospho-L-serine, ≥97% (TLC), from bovine brain, amorphous powder
Sigma-Aldrich
Cholesterol, Sigma Grade, ≥99%
Sigma-Aldrich
1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine, ≥97%
Sigma-Aldrich
Glyceryl tripalmitate, ≥99%
Sigma-Aldrich
L-α-Phosphatidylinositol from Glycine max (soybean), ~50% (TLC)
Sigma-Aldrich
1,2-Dipalmitoyl-sn-glycero-3-phosphate sodium salt, ≥99%
Sigma-Aldrich
Cholesteryl palmitate, ≥98% (HPLC; detection at 205 nm)
Sigma-Aldrich
Galactocerebrosides from bovine brain, ≥97% (TLC)