Protective effect of nilvadipine against glutamate neurotoxicity in purified retinal ganglion cells
We determined the effect of nilvadipine, a dihydropyridine-type calcium channel blocker, in preventing glutamate neurotoxicity in purified retinal ganglion cells (RGCs). RGCs were purified from dissociated rat retinal cells (postnatal days 6–8), using a modified two-step panning method, and cultured...
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| Veröffentlicht in: | Brain research 2003-01, Vol.961 (2), p.213-219 |
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| creator | Otori, Yasumasa Kusaka, Shunji Kawasaki, Atsushi Morimura, Hiroyuki Miki, Atsuya Tano, Yasuo |
| description | We determined the effect of nilvadipine, a dihydropyridine-type calcium channel blocker, in preventing glutamate neurotoxicity in purified retinal ganglion cells (RGCs). RGCs were purified from dissociated rat retinal cells (postnatal days 6–8), using a modified two-step panning method, and cultured in serum-free medium containing neurotrophic factors and forskolin. RGC survival after exposure to glutamate (25 μM) with nilvadipine or other calcium channel blockers was measured by calcein-acetoxymethyl ester staining after 3 days in culture. Changes in the level of intracellular Ca
2+ ([Ca
2+]
i) were measured with fura-2 fluorescence. Induction of apoptosis was evaluated using the TDT-dUTP terminal nick-end labeling technique. The neurotoxic effects of low doses of glutamate were blocked by a specific α-amino-3-dihydro-5-methylisoxazole-4-propionate-kainate receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (20 μM). Simultaneous application of nilvadipine (1–100 nM) with glutamate protected against glutamate neurotoxicity in a dose-dependent manner. Calcium-imaging experiments showed that the glutamate-evoked [Ca
2+]
i increase was significantly blocked by nilvadipine (
P |
| doi_str_mv | 10.1016/S0006-8993(02)03951-3 |
| format | Article |
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2+ ([Ca
2+]
i) were measured with fura-2 fluorescence. Induction of apoptosis was evaluated using the TDT-dUTP terminal nick-end labeling technique. The neurotoxic effects of low doses of glutamate were blocked by a specific α-amino-3-dihydro-5-methylisoxazole-4-propionate-kainate receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (20 μM). Simultaneous application of nilvadipine (1–100 nM) with glutamate protected against glutamate neurotoxicity in a dose-dependent manner. Calcium-imaging experiments showed that the glutamate-evoked [Ca
2+]
i increase was significantly blocked by nilvadipine (
P<0.001), but not nifedipine and diltiazem, in about 50% of RGCs. In addition, the application of nilvadipine significantly reduced glutamate-induced apoptosis (
P<0.001). These findings suggest that nilvadipine may partly inhibit glutamate-induced apoptotic cell death by blocking calcium influx via voltage-dependent calcium channels in purified RGCs.</description><identifier>ISSN: 0006-8993</identifier><identifier>EISSN: 1872-6240</identifier><identifier>DOI: 10.1016/S0006-8993(02)03951-3</identifier><identifier>PMID: 12531488</identifier><identifier>CODEN: BRREAP</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Animals ; Apoptosis ; Apoptosis - drug effects ; Biological and medical sciences ; Calcium - metabolism ; Calcium channel ; Calcium Channel Blockers - pharmacology ; Calcium Channels, L-Type - metabolism ; Cell Culture Techniques ; Cell Survival - drug effects ; Diltiazem - pharmacology ; Dose-Response Relationship, Drug ; Excitatory Amino Acid Antagonists - pharmacology ; Eye and associated structures. Visual pathways and centers. Vision ; Fundamental and applied biological sciences. Psychology ; Glutamate receptor ; Glutamic Acid - toxicity ; Neuroprotective Agents - pharmacology ; Nifedipine - analogs & derivatives ; Nifedipine - pharmacology ; Quinoxalines - pharmacology ; Rat ; Rats ; Receptors, AMPA - metabolism ; Receptors, Kainic Acid - metabolism ; Retinal cell culture ; Retinal ganglion cell ; Retinal Ganglion Cells - drug effects ; Retinal Ganglion Cells - metabolism ; Vertebrates: nervous system and sense organs</subject><ispartof>Brain research, 2003-01, Vol.961 (2), p.213-219</ispartof><rights>2002 Elsevier Science B.V.</rights><rights>2003 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c488t-aea2107bfc7b7aa6422e8b37de62869971ffcf7ad93d61076f2d8a288b25f5b83</citedby><cites>FETCH-LOGICAL-c488t-aea2107bfc7b7aa6422e8b37de62869971ffcf7ad93d61076f2d8a288b25f5b83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0006-8993(02)03951-3$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14467943$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12531488$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Otori, Yasumasa</creatorcontrib><creatorcontrib>Kusaka, Shunji</creatorcontrib><creatorcontrib>Kawasaki, Atsushi</creatorcontrib><creatorcontrib>Morimura, Hiroyuki</creatorcontrib><creatorcontrib>Miki, Atsuya</creatorcontrib><creatorcontrib>Tano, Yasuo</creatorcontrib><title>Protective effect of nilvadipine against glutamate neurotoxicity in purified retinal ganglion cells</title><title>Brain research</title><addtitle>Brain Res</addtitle><description>We determined the effect of nilvadipine, a dihydropyridine-type calcium channel blocker, in preventing glutamate neurotoxicity in purified retinal ganglion cells (RGCs). RGCs were purified from dissociated rat retinal cells (postnatal days 6–8), using a modified two-step panning method, and cultured in serum-free medium containing neurotrophic factors and forskolin. RGC survival after exposure to glutamate (25 μM) with nilvadipine or other calcium channel blockers was measured by calcein-acetoxymethyl ester staining after 3 days in culture. Changes in the level of intracellular Ca
2+ ([Ca
2+]
i) were measured with fura-2 fluorescence. Induction of apoptosis was evaluated using the TDT-dUTP terminal nick-end labeling technique. The neurotoxic effects of low doses of glutamate were blocked by a specific α-amino-3-dihydro-5-methylisoxazole-4-propionate-kainate receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (20 μM). Simultaneous application of nilvadipine (1–100 nM) with glutamate protected against glutamate neurotoxicity in a dose-dependent manner. Calcium-imaging experiments showed that the glutamate-evoked [Ca
2+]
i increase was significantly blocked by nilvadipine (
P<0.001), but not nifedipine and diltiazem, in about 50% of RGCs. In addition, the application of nilvadipine significantly reduced glutamate-induced apoptosis (
P<0.001). These findings suggest that nilvadipine may partly inhibit glutamate-induced apoptotic cell death by blocking calcium influx via voltage-dependent calcium channels in purified RGCs.</description><subject>Animals</subject><subject>Apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>Biological and medical sciences</subject><subject>Calcium - metabolism</subject><subject>Calcium channel</subject><subject>Calcium Channel Blockers - pharmacology</subject><subject>Calcium Channels, L-Type - metabolism</subject><subject>Cell Culture Techniques</subject><subject>Cell Survival - drug effects</subject><subject>Diltiazem - pharmacology</subject><subject>Dose-Response Relationship, Drug</subject><subject>Excitatory Amino Acid Antagonists - pharmacology</subject><subject>Eye and associated structures. Visual pathways and centers. Vision</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glutamate receptor</subject><subject>Glutamic Acid - toxicity</subject><subject>Neuroprotective Agents - pharmacology</subject><subject>Nifedipine - analogs & derivatives</subject><subject>Nifedipine - pharmacology</subject><subject>Quinoxalines - pharmacology</subject><subject>Rat</subject><subject>Rats</subject><subject>Receptors, AMPA - metabolism</subject><subject>Receptors, Kainic Acid - metabolism</subject><subject>Retinal cell culture</subject><subject>Retinal ganglion cell</subject><subject>Retinal Ganglion Cells - drug effects</subject><subject>Retinal Ganglion Cells - metabolism</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0006-8993</issn><issn>1872-6240</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU2LFDEQhoMo7rj6E5RcFD205qM7nT4tsvgFCwrqOVSnK0NJT3pM0oP7783sDO7RUxJ43srLU4w9l-KtFNK8-y6EMI0dBv1aqDdCD51s9AO2kbZXjVGteMg2_5AL9iTnX_Wp9SAeswupOi1bazfMf0tLQV_ogBxDqDe-BB5pPsBEe4rIYQsUc-HbeS2wg4I84lpDyx_yVG45Rb5fEwXCiScsFGHmW4jbmZbIPc5zfsoeBZgzPjufl-znxw8_rj83N18_fbl-f9P4WqU0gKCk6Mfg-7EHMK1SaEfdT2iUNcPQyxB86GEa9GQqaIKaLChrR9WFbrT6kr06zd2n5feKubgd5WMDiLis2UlrOtErU8HuBPq05JwwuH2iHaRbJ4U72nV3dt1RnRPK3dl1uuZenD9Yxx1O96mzzgq8PAOQPcwhQfSU77m2Nf3QHgddnTisOg6EyWVPGD1OlOoK3LTQf6r8BZDBmNY</recordid><startdate>20030131</startdate><enddate>20030131</enddate><creator>Otori, Yasumasa</creator><creator>Kusaka, Shunji</creator><creator>Kawasaki, Atsushi</creator><creator>Morimura, Hiroyuki</creator><creator>Miki, Atsuya</creator><creator>Tano, Yasuo</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TK</scope></search><sort><creationdate>20030131</creationdate><title>Protective effect of nilvadipine against glutamate neurotoxicity in purified retinal ganglion cells</title><author>Otori, Yasumasa ; Kusaka, Shunji ; Kawasaki, Atsushi ; Morimura, Hiroyuki ; Miki, Atsuya ; Tano, Yasuo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c488t-aea2107bfc7b7aa6422e8b37de62869971ffcf7ad93d61076f2d8a288b25f5b83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>Biological and medical sciences</topic><topic>Calcium - metabolism</topic><topic>Calcium channel</topic><topic>Calcium Channel Blockers - pharmacology</topic><topic>Calcium Channels, L-Type - metabolism</topic><topic>Cell Culture Techniques</topic><topic>Cell Survival - drug effects</topic><topic>Diltiazem - pharmacology</topic><topic>Dose-Response Relationship, Drug</topic><topic>Excitatory Amino Acid Antagonists - pharmacology</topic><topic>Eye and associated structures. Visual pathways and centers. Vision</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glutamate receptor</topic><topic>Glutamic Acid - toxicity</topic><topic>Neuroprotective Agents - pharmacology</topic><topic>Nifedipine - analogs & derivatives</topic><topic>Nifedipine - pharmacology</topic><topic>Quinoxalines - pharmacology</topic><topic>Rat</topic><topic>Rats</topic><topic>Receptors, AMPA - metabolism</topic><topic>Receptors, Kainic Acid - metabolism</topic><topic>Retinal cell culture</topic><topic>Retinal ganglion cell</topic><topic>Retinal Ganglion Cells - drug effects</topic><topic>Retinal Ganglion Cells - metabolism</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Otori, Yasumasa</creatorcontrib><creatorcontrib>Kusaka, Shunji</creatorcontrib><creatorcontrib>Kawasaki, Atsushi</creatorcontrib><creatorcontrib>Morimura, Hiroyuki</creatorcontrib><creatorcontrib>Miki, Atsuya</creatorcontrib><creatorcontrib>Tano, Yasuo</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><jtitle>Brain research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Otori, Yasumasa</au><au>Kusaka, Shunji</au><au>Kawasaki, Atsushi</au><au>Morimura, Hiroyuki</au><au>Miki, Atsuya</au><au>Tano, Yasuo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Protective effect of nilvadipine against glutamate neurotoxicity in purified retinal ganglion cells</atitle><jtitle>Brain research</jtitle><addtitle>Brain Res</addtitle><date>2003-01-31</date><risdate>2003</risdate><volume>961</volume><issue>2</issue><spage>213</spage><epage>219</epage><pages>213-219</pages><issn>0006-8993</issn><eissn>1872-6240</eissn><coden>BRREAP</coden><abstract>We determined the effect of nilvadipine, a dihydropyridine-type calcium channel blocker, in preventing glutamate neurotoxicity in purified retinal ganglion cells (RGCs). RGCs were purified from dissociated rat retinal cells (postnatal days 6–8), using a modified two-step panning method, and cultured in serum-free medium containing neurotrophic factors and forskolin. RGC survival after exposure to glutamate (25 μM) with nilvadipine or other calcium channel blockers was measured by calcein-acetoxymethyl ester staining after 3 days in culture. Changes in the level of intracellular Ca
2+ ([Ca
2+]
i) were measured with fura-2 fluorescence. Induction of apoptosis was evaluated using the TDT-dUTP terminal nick-end labeling technique. The neurotoxic effects of low doses of glutamate were blocked by a specific α-amino-3-dihydro-5-methylisoxazole-4-propionate-kainate receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (20 μM). Simultaneous application of nilvadipine (1–100 nM) with glutamate protected against glutamate neurotoxicity in a dose-dependent manner. Calcium-imaging experiments showed that the glutamate-evoked [Ca
2+]
i increase was significantly blocked by nilvadipine (
P<0.001), but not nifedipine and diltiazem, in about 50% of RGCs. In addition, the application of nilvadipine significantly reduced glutamate-induced apoptosis (
P<0.001). These findings suggest that nilvadipine may partly inhibit glutamate-induced apoptotic cell death by blocking calcium influx via voltage-dependent calcium channels in purified RGCs.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>12531488</pmid><doi>10.1016/S0006-8993(02)03951-3</doi><tpages>7</tpages></addata></record> |
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| subjects | Animals Apoptosis Apoptosis - drug effects Biological and medical sciences Calcium - metabolism Calcium channel Calcium Channel Blockers - pharmacology Calcium Channels, L-Type - metabolism Cell Culture Techniques Cell Survival - drug effects Diltiazem - pharmacology Dose-Response Relationship, Drug Excitatory Amino Acid Antagonists - pharmacology Eye and associated structures. Visual pathways and centers. Vision Fundamental and applied biological sciences. Psychology Glutamate receptor Glutamic Acid - toxicity Neuroprotective Agents - pharmacology Nifedipine - analogs & derivatives Nifedipine - pharmacology Quinoxalines - pharmacology Rat Rats Receptors, AMPA - metabolism Receptors, Kainic Acid - metabolism Retinal cell culture Retinal ganglion cell Retinal Ganglion Cells - drug effects Retinal Ganglion Cells - metabolism Vertebrates: nervous system and sense organs |
| title | Protective effect of nilvadipine against glutamate neurotoxicity in purified retinal ganglion cells |
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