Use of 2,3-Naphthalenedicarboxaldehyde Derivatization for Single-Cell Analysis of Glutathione by Capillary Electrophoresis and Histochemical Localization by Fluorescence Microscopy

Orwar, Owe; Fishman, Harvey A.; Ziv, Noam; Scheller, Richard H.; Zare, Richard N.

doi:10.1021/ac00119a010

Cited by 128 publications

(100 citation statements)

References 6 publications

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“…Reduced glutathione concentration in the tissues extracts was measured with 2,3-naphthalenedicarboxaldehyde derivatization as described in ref. 60.…”

Section: Methodsmentioning

confidence: 99%

β-Amyloid infusion results in delayed and age-dependent learning deficits without role of inflammation or β-amyloid deposits

Malm¹,

Ort

Tähtivaara

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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␤-Amyloid (A␤) polypeptide plays a critical role in the pathogenesis of Alzheimer's disease (AD), which is characterized by progressive decline of cognitive functions, formation of A␤ deposits and neurofibrillary tangles, and loss of neurons. Increased genetic production or direct intracerebral administration of A␤ in animal models results in A␤ deposition, gliosis, and impaired cognitive functions. Whether aging renders the brain prone to A␤ and whether inflammation is required for A␤-induced learning deficits is unclear. We show that intraventricular infusion of A␤ 1-42 results in learning deficits in 9-month-old but not 2.5-month-old mice. Deficits that become detectable 12 weeks after the infusion are associated with a slight reduction in Cu,Zn superoxide dismutase activity but do not correlate with A␤ deposition and are not associated with gliosis. In rats, A␤ infusion induced learning deficits that were detectable 6 months after the infusion. Approximately 20% of the A␤ immunoreactivity in rats was associated with astrocytes. NMR spectrum analysis of the animals cerebrospinal fluid revealed a strong reduction trend in several metabolites in A␤-infused rats, including lactate and myo-inositol, supporting the idea of dysfunctional astrocytes. Even a subtle increase in brain A␤ 1-42 concentration may disrupt normal metabolism of astrocytes, resulting in altered neuronal functions and age-related development of learning deficits independent of A␤ deposition and inflammation.aging ͉ cerebrospinal fluid ͉ learning ͉ oxidative stress ͉ Alzheimer's disease A lzheimer's disease (AD) is the most common cause of progressive impairment of cognitive functions in humans during aging (1). Diagnosis is confirmed by the presence of neurofibrillary tangles (formed via hyperphosphorylation of tau) and ␤-amyloid (A␤)-containing plaques (2). A␤, a 39-to 43-aalong polypeptide derived from the A␤ precursor protein (APP), is neurotoxic and plays a critical role in the pathogenesis of AD (1, 2). Mutations in APP or presenilin genes that lead to the elevation of fibrillogenic form of A␤ are responsible for only a small portion of AD cases (1,3,4). In addition, triplication of APP gene-containing chromosome 21 in Down's syndrome results in AD, and the transgenic (TG) mouse models (5) overexpressing human mutant APP or presenilin genes severalfold, show progressive accumulation of A␤ deposits, gliosis, and cognitive decline (2). The relationship between A␤ deposition and cognitive decline is unclear because the severity of dementia does not correlate with A␤ plaques (6). In TG models, cognitive deficits can occur in the absence of A␤ deposits (7-10). Recent findings emphasize the role of soluble A␤ oligomers. A␤ oligomers are highly neurotoxic (11), and their amount in the brain correlates with the severity of AD and with spatial learning deficits in APP TG mice (12). However, it is not completely clear whether the neuropathology in these mice is solely a consequence of A␤ accumulation or whether increased concentrations of other cle...

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“…Reduced glutathione concentration in the tissues extracts was measured with 2,3-naphthalenedicarboxaldehyde derivatization as described in ref. 60.…”

Section: Methodsmentioning

confidence: 99%

β-Amyloid infusion results in delayed and age-dependent learning deficits without role of inflammation or β-amyloid deposits

Malm¹,

Ort

Tähtivaara

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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show abstract

“…Reduced glutathione (GSH) concentration in the tissue extracts was measured with 2,3-naphthalenedicarboxaldehyde derivatization as described previously (Orwar et al, 1995). Superoxide dismutase 1 (SOD1) activity was measured from tissue homogenates by a previously described method (Beauchamp and Fridovich, 1971).…”

Section: Measurement Of Copper Concentration and Markers Of Oxidativementioning

confidence: 99%

Pyrrolidine Dithiocarbamate Activates Akt and Improves Spatial Learning in APP/PS1 Mice without Affecting β-Amyloid Burden

Malm¹,

Iivonen²,

Goldsteins³

et al. 2007

J. Neurosci.

146

108

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Pyrrolidine dithiocarbamate (PDTC) is a clinically tolerated inhibitor of nuclear factor-B (NF-B), antioxidant and antiinflammatory agent, which provides protection in brain ischemia models. In neonatal hypoxia-ischemia model, PDTC activates Akt and reduces activation of glycogen synthase kinase 3␤ (GSK-3␤). Because chronic inflammation, oxidative stress, and increased GSK-3␤ activity are features of Alzheimer's disease (AD) pathology, we tested whether PDTC reduces brain pathology and improves cognitive function in a transgenic animal model of AD. A 7 month oral treatment with PDTC prevented the decline in cognition in AD mice without altering ␤-amyloid burden or gliosis. Moreover, marked oxidative stress and activation of NF-B were not part of the brain pathology. Instead, the phosphorylated form of GSK-3␤ was decreased in the AD mouse brain, and PDTC treatment increased the phosphorylation of Akt and GSK-3␤. Also, PDTC treatment increased the copper concentration in the brain. In addition, PDTC rescued cultured hippocampal neurons from the toxicity of oligomeric A␤ and reduced tau phosphorylation in the hippocampus of AD mice. Finally, astrocytic glutamate transporter GLT-1, known to be regulated by Akt pathway, was decreased in the transgenic AD mice but upregulated back to the wild-type levels by PDTC treatment. Thus, PDTC may improve spatial learning in AD by interfering with Akt-GSK pathway both in neurons and astrocytes. Because PDTC is capable of transferring external Cu 2ϩ into a cell, and, in turn, Cu 2ϩ is able to activate Akt, we hypothesize that PDTC provides the beneficial effect in transgenic AD mice through Cu 2ϩ -activated Akt pathway.

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“…In the presence of ROS, similar to those generated by radiation, GSH neutralises the radicals, undergoing oxidation primarily to form a glutathione disulphide dimer (GSSG). The monomer and oxidised dimer can be separated by CE (18) and fluorescently detected. In this case, derivatization should be done following the separation (22) as GSSG does not have a free thiol group that can be labelled.…”

Section: Fluorescence Detectionmentioning

confidence: 99%

“…(14) Nitric oxide (NO † ) DAF-FM 495 515 (13) Hydroxyl (OH † ) TA 315 425 (12) 'General' ROS DHR123, DHE, MitoSOX 505 500 525 600 (9 -11) Glutathione (GSH, GSSG) NDA, OPA 430 338 530 458 (18,19) Figure 1. Fluorescence spectra measured using the RARAF breadboard LIF system of DHR-123 before (grey) and after (black) addition of KO 2 , a superoxide generator.…”

Section: Coupling Ce-lif To a Microbeammentioning

confidence: 99%

Microbeam-coupled capillary electrophoresis

Garty

Ehsan

Yang

et al. 2015

Radiat Prot Dosimetry

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Use of 2,3-Naphthalenedicarboxaldehyde Derivatization for Single-Cell Analysis of Glutathione by Capillary Electrophoresis and Histochemical Localization by Fluorescence Microscopy

Cited by 128 publications

References 6 publications

β-Amyloid infusion results in delayed and age-dependent learning deficits without role of inflammation or β-amyloid deposits

β-Amyloid infusion results in delayed and age-dependent learning deficits without role of inflammation or β-amyloid deposits

Pyrrolidine Dithiocarbamate Activates Akt and Improves Spatial Learning in APP/PS1 Mice without Affecting β-Amyloid Burden

Microbeam-coupled capillary electrophoresis

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