Proteomic analysis of the brain tissues from a transgenic mouse model of amyloid β oligomers

Takano, Masaoki; Matsushima, Kouji; Otani, Mieko; Sano, Keiji; Nakamura-Hirota, Tooru; Tokuyama, Shogo; Min, Kyong Son; Tomiyama, Takami; Mori, Hiroshi; Matsuyama, Shogo

doi:10.1016/j.neuint.2012.05.018

Cited by 20 publications

(23 citation statements)

References 29 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%

“…29,30) The samples were dissolved in 5 µL of 50% ACN/0.3% (v/v) trifluoroacetic acid. For the matrix, α-cyano-4-hydroxycinnamic acid (1 µg/µL: Wako Junyaku, Osaka, Japan) dissolved in the same mixture was used.…”

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%

Section: Methodsmentioning

confidence: 99%

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