Proteomic analysis of the hippocampus in Alzheimer's disease model mice by using two-dimensional fluorescence difference in gel electrophoresis

Takano, Masaoki; Yamashita, Takuya; Nagano, Kazuya; Otani, Mieko; Matsushima, Kouji; Kamada, Haruhiko; Tsunoda, Shin-ichi; Tsutsumi, Yasuo; Tomiyama, Takami; Mori, Hiroshi; Matsuura, Kenji; Matsuyama, Shogo

doi:10.1016/j.neulet.2012.11.010

Cited by 38 publications

(38 citation statements)

References 26 publications

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“…29,30) Protein (300 µg) was applied to ImmobilineDryStrip pH 3-10 NL (7 cm) gels (GE Healthcare UK Ltd.) and separated at 50 V for 6 h, at 100 V for 6 h, and at 2000 V for 6 h. The IPG strips were equilibrated for 15 min in 50 mM Tris-HCl (pH 8.8), 6 M urea, 30% (v/v) glycerol, 1% SDS, and 1% (v/v) dithiothreitol (DTT), and then for 15 min in the same buffer with 2.5% (w/v) iodoacetamide instead of DTT. After equilibration, the IPG strips were placed onto a 12.5% acrylamide gel and SDS-PAGE was performed at 5 mA/ gel for 7 h. SYPRO Ruby Staining Proteins on the SDS-polyacrylamide gels were detected using SYPRO Ruby Protein Gel Stain (Molecular Probes) as previously described, with some modification.…”

Section: Methodsmentioning

confidence: 99%

“…After equilibration, the IPG strips were placed onto a 12.5% acrylamide gel and SDS-PAGE was performed at 5 mA/ gel for 7 h. SYPRO Ruby Staining Proteins on the SDS-polyacrylamide gels were detected using SYPRO Ruby Protein Gel Stain (Molecular Probes) as previously described, with some modification. 29,30) Gels after 2-DE were fixed in a solution containing 10% acetic acid/50% methanol for 30 min, then 7% acetic acid/10% methanol for 30 min. After fixation, the gels were incubated in undiluted stock solution of SYPRO Ruby for 90 min and then destained with 7% acetic acid/10% methanol for 30 min.…”

Section: Methodsmentioning

confidence: 99%

“…29,30) Each of three sets of samples was represented by two independent biological replicates of 2-DE gels. The gel images were normalized with the Prodigy SameSpots software to compensate for differences in the staining intensity among the gels.…”

Section: Methodsmentioning

confidence: 99%

“…29,30) Protein spots in the gels stained with SYPRO Ruby was cut out and subjected to trypsin digestion with porcine trypsin (Promega, Madison, WI, U.S.A.). Briefly, gel pieces were washed, dehydrated, and diluted in 200 µL of 25 mM ammonium hydrogen carbonate with 5% acetonitrile (ACN) (v/v).…”

Section: Methodsmentioning

confidence: 99%

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Withdrawal: Proteomic Profiling in the Spinal Cord and Sciatic Nerve in a Global Cerebral Ischemia-Induced Mechanical Allodynia Mouse Model

Harada

Matsuura

Takano

et al. 2016

Biological & Pharmaceutical Bulletin

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Central post-stroke pain (CPSP) is one of the complications of cerebral ischemia and neuropathic pain syndrome. At present, there are few studies of pain in regions such as the spinal cord or sciatic nerve in cerebral ischemic animal models. To identify proteomic changes in the spinal cord and sciatic nerve in global cerebral ischemic model mice, in the present study we performed an investigation using proteomic methods. In a comparison between the intensity of protein spots obtained from a sham and that from a bilateral carotid artery occulusion (BCAO) in spinal cord and sciatic nerve, the levels of 10 (spinal cord) and 7 (sciatic nerve) protein spots were altered. The protein levels in the spinal cord were significantly increased in N G ,N Gdimethylarginine dimethylaminohydrolase 1 (DDAH1), 6-phosphogluconolactonase isoform 1, and precursor apoprotein A-I and decreased in dihydropyrimidinase-related protein 2 (CRMP-2), enolase 1B, rab guanosine 5′-diphosphate (GDP) dissociation inhibitor beta, septin-2 isoform a, isocitrate dehydrogenase subunit alpha, cytosolic malate dehydrogenase, and adenosine triphosphate synthase. The protein levels in the sciatic nerve were significantly increased in a mimecan precursor, myosin light chain 1/3, and myosin regulatory light chain 2 (MLC2), and decreased in dihydropyrimidinase-related protein 3 (CRMP-4), protein disulfideisomerase A3, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 1, and B-type creatine kinase. In addition, CRMP-2 and CRMP-4 protein levels were decreased, and DDAH1 and MLC2 protein levels were increased on day 1 after BCAO using Western blotting. These results suggested that changes in these proteins may be involved in the regulation of CPSP.Key words global ischemia; pain; spinal cord; sciatic nerve; proteome Cerebral stroke remains one of the leading causes of death and disability worldwide.1) Recent insights into the basic mechanism involved in ischemic stroke indicate that endothelial dysfunctions along with oxidative stress and neuroinflammation are key elements in the occurrence and development of ischemic brain damage that results in cell damage and death.2,3) Many clinical symptoms can manifest after strokes, including motor deficits, cognitive dysfunction, language problems, emotional disturbances, social maladjustment, somatosensory dysfunction, and central pain. 4,5) Cerebral stroke occasionally induces central post-stroke pain (CPSP) that manifests as burning pain, throbbing pain, aching pain, slashing pain, allodynia, and hyperalgesia, either continuously or intermittently on the affected side.6,7) CPSP is known to be one of the neuropathic pain.8) Neuropathic pain is defined as "pain initiated or caused by a primary lesion or dysfunction of the nervous system." It can be a debilitating and difficult condition to treat and is often resistant to simple analgesics, requiring additional analgesic treatment. 9)Some hypotheses of the pathophysiological mechanisms of somatosensory dysfunction have been proposed to explain CPSP symptoms, such as the ...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Withdrawal: Proteomic Profiling in the Spinal Cord and Sciatic Nerve in a Global Cerebral Ischemia-Induced Mechanical Allodynia Mouse Model

Harada

Matsuura

Takano

et al. 2016

Biological & Pharmaceutical Bulletin

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“…PACSIN1 is a neuronal-specific protein that regulates activitydependent retrieval of synaptic vesicles in the presynaptic terminals (19)(20)(21)(22) as well as the endocytosis of the NR3A subunit of NMDA receptors on the postsynaptic membrane (23). More importantly, PACSIN1 has been implicated in various neurodegenerative disorders, including Huntington and Alzheimer's diseases (24,25). PACSIN2 is ubiquitously expressed, whereas the expression of PACSIN3 is restricted to the muscle, heart, and lung (26).…”

Section: Syndapin | Receptor Traffickingmentioning

confidence: 99%

PICK1 interacts with PACSIN to regulate AMPA receptor internalization and cerebellar long-term depression

Anggono

Koç-Schmitz

Widagdo

et al. 2013

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

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The dynamic trafficking of AMPA receptors (AMPARs) into and out of synapses is crucial for synaptic transmission, plasticity, learning, and memory. The protein interacting with C-kinase 1 (PICK1) directly interacts with GluA2/3 subunits of the AMPARs. Although the role of PICK1 in regulating AMPAR trafficking and multiple forms of synaptic plasticity is known, the exact molecular mechanisms underlying this process remain unclear. Here, we report a unique interaction between PICK1 and all three members of the protein kinase C and casein kinase II substrate in neurons (PACSIN) family and show that they form a complex with AMPARs. Our results reveal that knockdown of the neuronal-specific protein, PACSIN1, leads to a significant reduction in AMPAR internalization following the activation of NMDA receptors in hippocampal neurons. The interaction between PICK1 and PACSIN1 is regulated by PACSIN1 phosphorylation within the variable region and is required for AMPAR endocytosis. Similarly, the binding of PICK1 to the ubiquitously expressed PACSIN2 is also regulated by the homologous phosphorylation sites within the PACSIN2-variable region. Genetic deletion of PACSIN2, which is highly expressed in Purkinje cells, eliminates cerebellar long-term depression. This deficit can be fully rescued by overexpressing wild-type PACSIN2, but not by a PACSIN2 phosphomimetic mutant, which does not bind PICK1 efficiently. Taken together, our data demonstrate that the interaction of PICK1 and PACSIN is required for the activity-dependent internalization of AMPARs and for the expression of long-term depression in the cerebellum.syndapin | receptor trafficking

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Pharmacogenomics of Alzheimer’s Disease: Novel Therapeutic Strategies for Drug Development

Cacabelos

Torrellas

et al. 2014

Methods in Molecular Biology

165

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Alzheimer's disease (AD) is a major problem of health and disability, with a relevant economic impact on our society. Despite important advances in pathogenesis, diagnosis, and treatment, its primary causes still remain elusive, accurate biomarkers are not well characterized, and the available pharmacological treatments are not cost-effective. As a complex disorder, AD is a polygenic and multifactorial clinical entity in which hundreds of defective genes distributed across the human genome may contribute to its pathogenesis. Diverse environmental factors, cerebrovascular dysfunction, and epigenetic phenomena, together with structural and functional genomic dysfunctions, lead to amyloid deposition, neurofibrillary tangle formation, and premature neuronal death, the major neuropathological hallmarks of AD. Future perspectives for the global management of AD predict that genomics and proteomics may help in the search for reliable biomarkers. In practical terms, the therapeutic response to conventional drugs (cholinesterase inhibitors, multifactorial strategies) is genotype-specific. Genomic factors potentially involved in AD pharmacogenomics include at least five categories of gene clusters: (1) genes associated with disease pathogenesis; (2) genes associated with the mechanism of action of drugs; (3) genes associated with drug metabolism (phase I and II reactions); (4) genes associated with drug transporters; and (5) pleiotropic genes involved in multifaceted cascades and metabolic reactions. The implementation of pharmacogenomic strategies will contribute to optimize drug development and therapeutics in AD and related disorders.

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Proteomic analysis of the hippocampus in Alzheimer's disease model mice by using two-dimensional fluorescence difference in gel electrophoresis

Cited by 38 publications

References 26 publications

Withdrawal: Proteomic Profiling in the Spinal Cord and Sciatic Nerve in a Global Cerebral Ischemia-Induced Mechanical Allodynia Mouse Model

Withdrawal: Proteomic Profiling in the Spinal Cord and Sciatic Nerve in a Global Cerebral Ischemia-Induced Mechanical Allodynia Mouse Model

PICK1 interacts with PACSIN to regulate AMPA receptor internalization and cerebellar long-term depression

Pharmacogenomics of Alzheimer’s Disease: Novel Therapeutic Strategies for Drug Development

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