Different trends in modulation of NMDAR1 and NMDAR2B gene expression in cultured cortical and hippocampal neurons after lead exposure

Lau, W.S.; Yeung, C.W.; Lui, P.W.; Cheung, L.H.; Poon, Nick; Yung, Kin Lam

doi:10.1016/s0006-8993(01)03395-9

Cited by 38 publications

(16 citation statements)

References 39 publications

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“…Western blot analysis was performed in accordance with a previous study [39] . Thirty hours after the oligo treatments, neurons in culture were washed (3 !…”

Section: Western Blot Analysismentioning

confidence: 99%

Ablation of Gene Expression ofN-Methyl-<i>D</i>-Aspartate Receptor One by Antisense Oligonucleotides in Striatal Neurons in Culture

Lui

Yeung²,

Yung³

et al. 2005

Neurosignals

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In the present study, a twenty-mer antisense oligonucleotide specific for N-methyl-D-aspartate receptor one (ANR1) was applied to striatal neurons in primary cell culture. The ANR1 was found to be specific and nontoxic. Significant reductions in expression of NR1 mRNA and proteins were resulted after a single dose of ANR1 transcripts. Interestingly, there were reductions in total NR1 proteins but two phosphorylated forms of NR1 proteins at serine 896 and 897 residues were not reduced. There was also no change in the pattern of distribution of NR1 immunoreactivity in the striatal neurons. In addition, significant reductions of NMDA-mediated peak inward current were found after application of a higher concentration of ANR1 (20–100 µM) by patch clamp recordings. The present results indicate that ANR1 is a useful agent in reducing NMDA receptor functions. The present data thus provide detailed cellular and molecular mechanisms to explain our previous findings of amelioration of motor symptoms in a rat model of Parkinson’s disease. More importantly, application of ANR1 was also found to display neuroprotective effects of striatal neurons against NMDA-induced excitotoxic cell death. The findings have implications in development of new approach in prevention of cell death in neurodegenerative diseases and new treatments for these diseases.

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“…Western blot analysis was performed in accordance with a previous study [39] . Thirty hours after the oligo treatments, neurons in culture were washed (3 !…”

Section: Western Blot Analysismentioning

confidence: 99%

Ablation of Gene Expression ofN-Methyl-<i>D</i>-Aspartate Receptor One by Antisense Oligonucleotides in Striatal Neurons in Culture

Lui

Yeung²,

Yung³

et al. 2005

Neurosignals

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“…The polymerase chain reaction (PCR) was carried out with Gene Taq (Nippon Gene, Co., Ltd., Tokyo). The oligonucleotide primers for NR2B were designed based on the sequences described by Lau et al (12). The sequences of PCR primers were as follows: NR2B, 5′-TCC GTC TTT CTT ATG GG ATA TGC-3′ (sense), 5′-CCT CTA GGC GGA CAG ATT AAG G-3′ (antisense); glyceraldehyde-3-phosphate dehydrogenase (G3PDH), 5′-YGC CTG CTT CAC CAC CTT-3′ (sense), 5′-TGC MTC CTG CAC CAC CAA CT-3′ (antisense) (Sigma-Aldrich, St. Louis, MO, USA).…”

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confidence: 99%

Repeated Administration of Amitriptyline Reduces Oxaliplatin-Induced Mechanical Allodynia in Rats

et al. 2012

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Oxaliplatin has widely been used as a key drug in the treatment of colorectal cancer. However, it causes severe acute and chronic peripheral neuropathies. Acute neuropathy includes acral paresthesias and dysesthesia triggered or enhanced by exposure to cold, and it appears soon after administration (1). After multiple cycles of treatment, the patients develop chronic neuropathy characterized by sensory and motor dysfunction. This chronic neuropathy is a dose-limiting toxicity and a major clinical problem in oxaliplatin chemotherapy (2).Antidepressant drugs have been recommended to be used as first-line drugs for the treatment of neuropathic pain (3,4), and in particular, amitriptyline is demonstrated to possess an analgesic activity for neuropathic pain in many randomized controlled trials (5).Recently, we reported that repeated administration of oxaliplatin induced cold hyperalgesia in the early phase and mechanical allodynia in the late phase in rats (6), and spinal NR2B-containing N-methyl-D-aspartate (NMDA) receptors are involved in the oxaliplatin-induced mechanical allodynia (7). However, the effect of amitriptyline on the oxaliplatin-induced neuropathy remains unexplored. Accordingly, we investigated the effect of amitriptyline on the oxaliplatin-induced cold hyperalgesia and mechanical allodynia in rats. Moreover, we examined the effect of amitriptyline on the oxaliplatininduced increase in the expression of NR2B protein and mRNA in the rat spinal cord.Male Sprague-Dawley rats (Kyudo Co., Tosu) were used. Rats were housed in groups of four to five per cage, with lights on from 7:00 AM to 7:00 PM. Animals had free access to food and water in their home cages. All experiments were approved by the Experimental Animal Care and Use Committee of Kyushu University ( Fukuoka) according to the National Institutes of Health guidelines, and we followed the International Association for the Study of Pain (IASP) Committee for Research and Ethical Issues guidelines for animal research (8).Oxaliplatin (Elplat ® ) was obtained from Yakult Co., Ltd. (Tokyo) and was dissolved in 5% glucose solution. Amitriptyline was provided by Wako Pure Chemical Industries, Ltd. (Osaka) and was dissolved in distilled water. Oxaliplatin (4 mg/kg) or vehicle (5% glucose solution) was injected intraperitoneally (i.p.) in volumes of 1 mL/kg, twice a week for 4 weeks (days 1, 2, 8, 9, 15, 16, 22, and 23). Amitriptyline (5 and 10 mg/kg) was administered p.o. once a day for 27 days (from day 1). The doses of these drugs were chosen based on previous reports (6,7,9). Behavioral tests were performed blindly with respect to drug administration.The cold hyperalgesia was assessed by acetone test described by Flatters and Bennett (10). The acetone test was performed before the first drug administration (on day 0) and on days 4, 7, 11, 14, 18, 21, and Pharmacy, Kyushu University Hospital, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan Received January 6, 2012; Accepted February 17, 2012 Abstract. Oxaliplatin is a key drug in the treatme...

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“…In contrast to NR1, which is expressed constitutively, NMDA receptor R2B (NR2B) expression is highly regulated during development of the brain. Studies have revealed that exposure to drugs such dioxin (Kakeyama et al, 2001), NMDA antagonists MK-801 and activator protein-5 , ethanol (Kalluri et al, 1998;Kumari and Ticku, 1998), and heavy metal lead (Lau et al, 2002) altered the expression of the NR2B gene. Transcription regulation of NMDA receptor components has been described in more detail for the NR1 subunit.…”

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confidence: 99%

Neuron-Restrictive Silencer Factor Regulates theN-Methyl-d-aspartate Receptor 2B Subunit Gene in Basal and Ethanol-Induced Gene Expression in Fetal Cortical Neurons

Mei

Rani²,

Ticku

2005

Mol Pharmacol

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Neuron-restrictive silencer factor (NRSF) is a transcriptional repressor of multiple neuronal genes. This study addressed the role of NRSF in N-methyl-D-aspartate (NMDA) receptor NR2B promoter activity and the molecular mechanisms of ethanolinduced NR2B up-regulation in fetal cortical neurons. The 5Ј-flanking region of the NR2B gene contains five NRSE-like elements. Functional analysis of the upstream regions of the NR2B gene by transient transfection of neurons revealed that neuronrestrictive silencer element (NRSE) motifs located between base pair Ϫ1407 and Ϫ2741 represses transcription of the gene. Analysis by electrophoretic mobility shift assay and reporter gene assay identified NRSE2 and 3 as responsible for repressing NR2B gene transcription. The identity of NRSF as the functional binding factor is suggested by the specific binding of in vitro synthesized NRSF or cell lysate to the labeled probes and the specific antibody-induced supershift. Furthermore, whereas mutations of NRSE2 and 3 motifs increased the promoter activity, overexpression of NRSF reduced it significantly. The pattern of NRSF expression during development was investigated and demonstrated that the highest expression is on embryonic day 14 with moderate expression on postnatal day 0, reflecting a possible role of NRSF as a regulator during development. Treatment of cultured cortical neurons with 100 mM ethanol for 5 days caused a significant decrease in the NRSF mRNA and protein levels, NRSF/NRSE binding activity, and an increase in the promoter activity. Therefore, our studies suggest that NRSF is a negative regulator of NR2B expression and may contribute to the ethanol-induced up-regulation of the NR2B gene in fetal cortical neurons.The neuron-restrictive silencer element (NRSE), also called repressor element-1, has been identified in many genes. Studies indicate that NRSE mediates transcriptional repression of many neuron-specific genes via the neuronrestrictive silencer factor (NRSF) or repressor element silencing transcription factor (Chong et al

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Different trends in modulation of NMDAR1 and NMDAR2B gene expression in cultured cortical and hippocampal neurons after lead exposure

Cited by 38 publications

References 39 publications

Ablation of Gene Expression ofN-Methyl-<i>D</i>-Aspartate Receptor One by Antisense Oligonucleotides in Striatal Neurons in Culture

Ablation of Gene Expression ofN-Methyl-<i>D</i>-Aspartate Receptor One by Antisense Oligonucleotides in Striatal Neurons in Culture

Repeated Administration of Amitriptyline Reduces Oxaliplatin-Induced Mechanical Allodynia in Rats

Neuron-Restrictive Silencer Factor Regulates theN-Methyl-d-aspartate Receptor 2B Subunit Gene in Basal and Ethanol-Induced Gene Expression in Fetal Cortical Neurons

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