Development and Validation of an HPTLC Method for Simultaneous Estimation of Excitatory Neurotransmitters in Rat Brain

Babu, Chidambaram Saravana; Sunil, Adoor Gopalan; Vasanthi, Hannah R.; Muthusamy, Velusamy Shanmuganathan; Ramanathan, Meena

doi:10.1080/10826070701588760

Cited by 19 publications

(8 citation statements)

References 10 publications

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“…However, high levels of glutamate show excitotoxicity, which is a critical mechanism contributing to neuronal death [ 31 ]. Several studies showed that the concentration of glutamate continued to be elevated in ischemia and dementia [ 12 , 32 , 33 ]. The high level of glutamate in hippocampus of VD rats was observed in our experiment, which indicated that VD was able to disturb the glutamate level of rats and led to excitotoxicity.…”

Section: Discussionmentioning

confidence: 99%

“…However, high levels of glutamate is neurotoxic [ 11 ]. Studies show that the concentration of glutamate continues to be elevated in dementia and cerebral ischemia [ 7 , 12 ], which leads to the excessive flux of calcium inside the neuron. Then, neuronal homeostasis is severely impaired and eventually, neurodegeneration happens [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

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Hydrogen Sulfide Prevents Synaptic Plasticity from VD-Induced Damage via Akt/GSK-3β Pathway and Notch Signaling Pathway in Rats

Liu

Gao

et al. 2015

Mol Neurobiol

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Our previous study has demonstrated that hydrogen sulfide (H2S) attenuates neuronal injury induced by vascular dementia (VD) in rats, but the mechanism is still poorly understood. In this study, we aimed to investigate whether the neuroprotection of H2S was associated with synaptic plasticity and try to interpret the potential underlying mechanisms. Adult male Wistar rats were suffered the ligation of bilateral common carotid arteries. At 24 h after surgery, rats were administered intraperitoneally with sodium hydrosulfide (NaHS, 5.6 mg·kg−1·day−1), a H2S donor, for 3 weeks in the VD+NaHS group and treated intraperitoneally with saline in the VD group respectively. Our results demonstrated that NaHS significantly decreased the level of glutamate. It obviously ameliorated cognitive flexibility as well as the spatial learning and memory abilities by Morris water maze. Moreover, NaHS significantly improved the long-term depression (LTD), and was able to elevate the expression of N-methyl-d-aspartate receptor subunit 2A, which plays a pivotal role in synaptic plasticity. Interestingly, NaHS decreased the phosphorylation of Akt, and it could maintain the activity of glycogen synthase kinase-3β (GSK-3β). Surprisingly, NaHS triggered the canonical Notch pathway by increasing expressions of Jagged-1 and Hes-1. These findings suggest that NaHS prevents synaptic plasticity from VD-induced damage partly via Akt/GSK-3β pathway and Notch signaling pathway.Hydrogen sulfide modulated the ratio of NMDAR 2A/2B and improved the synaptic plasticity via Akt/GSK-3β pathway and Notch signaling pathway in VD rats.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Hydrogen Sulfide Prevents Synaptic Plasticity from VD-Induced Damage via Akt/GSK-3β Pathway and Notch Signaling Pathway in Rats

Liu

Gao

et al. 2015

Mol Neurobiol

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“…Purity of GABA in the brain tissue sample was confirmed by comparing the UV absorption spectra at the start, middle, and end position of the bands. Based on our earlier report [19], we fixed the mobile phase: n-butanol: glacial acetic and water (65 : 18 : 28 v/v/v) with dual development procedure for achieving the best resolution of GABA (R F -0.49). GABA was visualized and detected by derivatizing the plate in 0.2% w/v ninhydrin after the second run.…”

Section: Resultsmentioning

confidence: 99%

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

Babu

Kesavanarayanan

Kalaivani

et al. 2011

Chromatography Research International

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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 thinlayer 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.

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“…HPTLC (CAMAG, version 1.3.4, USA) chromatographic condition: Silica gel GF 254 as Stationary phase; n‐butanol:glacial acetic acid:water (65:15:25, v/v/v) as mobile phase; Applicator: Linomat V; Scanner: CAMAG TLC scanner III; Developing chamber: twin trough glass chamber (20 × 10); Developing mode: ascending mode (multiple development); Detection reagent: 0.2% ninhydrin in acetone; scanning wavelength: 486 nm; experimental condition: 25 ± 2 °C temp/RH: 30–65%. Standard solutions of l ‐glutamic acid were prepared at 20–200 ng for plotting calibration curve (Babu et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

Post‐ischemic administration of nimodipine following focal cerebral ischemic‐reperfusion injury in rats alleviated excitotoxicity, neurobehavioural alterations and partially the bioenergetics

Babu

Ramanathan²

2010

Intl J of Devlp Neuroscience

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The present study focuses on the temporal calcium significance in middle cerebral artery occluded (2 h ischemia)-reperfused (70 h reperfusion) rats treated with nimodipine (NM) through concurrent measurements of excitotoxicity, bioenergetics and neurobehavioural paradigms. Further, the suitable therapeutic time window of calcium channel antagonism in stroke was also ascertained. NM (5 mg/kg, i.p.) was administered at pre (30 min before the induction of ischemia), during (1 h following occlusion of MCA) and post-ischemic (3 h after begin of reperfusion) states. The magnitude of neuroprotection in terms of excitotoxicity (glutamate, glutamine synthetase, Na(+)K(+)ATPase), bioenergetics (ATP, NAD(+)) and neurobehavioural paradigms (neurological score and open field exploratory behaviour) were measured and compared to ensure the therapeutic time-window of NM in stroke. Middle cerebral artery occlusion-reperfusion (MCAO/R) was found to elevate glutamate, glutamine synthetase levels and deplete Na(+)K(+)ATPase activity in the vehicle treated group (IR group). Significant decrease in bioenergetics such as ATP and NAD(+) levels was also observed. Further, IR group demonstrated grievous oxidative stress (increase in lipid peroxidation, protein carbonyl content, nitrite/nitrate levels and decrease in superoxide dismutase and glutathione levels) along with anxiogenic behaviour, neurological deficits and neuronal damage and decreased nuclear to cytoplasm ratio in CA1 hippocampal region. Post-ischemic NM administration reversed the excitotoxicity, neurobehavioural and histopathological alterations significantly, but it restored bioenergetics level in MCAO/R rats only partially. These findings were further confirmed with the combination treatment (CT) of post-ischemic NM and pre-ischemic memantine (MN) administration, since MN showed protective effect in the pre-ischemic administration (Babu and Ramanathan, 2009). The failure of NM to forefend the neurodegeneration on pre- and during-ischemic administration suggests that the initial phase damages in ischemic-reperfusion (IR) might be mediated through other mechanism(s) such as glutamergic overstimulation or reverse operation of glutamate transporters. From the present study, it is concluded that calcium plays a crucial role in post-ischemic status and the suitable therapeutic time window of calcium antagonism is the post-ischemic state.

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Development and Validation of an HPTLC Method for Simultaneous Estimation of Excitatory Neurotransmitters in Rat Brain

Cited by 19 publications

References 10 publications

Hydrogen Sulfide Prevents Synaptic Plasticity from VD-Induced Damage via Akt/GSK-3β Pathway and Notch Signaling Pathway in Rats

Hydrogen Sulfide Prevents Synaptic Plasticity from VD-Induced Damage via Akt/GSK-3β Pathway and Notch Signaling Pathway in Rats

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

Post‐ischemic administration of nimodipine following focal cerebral ischemic‐reperfusion injury in rats alleviated excitotoxicity, neurobehavioural alterations and partially the bioenergetics

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