Foreign metallothionein-I expression by transient transfection in MT-I and MT-II null astrocytes confers increased protection against acute methylmercury cytotoxicity
The mechanisms associated with metallothionein (MT) gene regulation are complex and poorly understood. Only a modest increase in brain MT expression levels is attained by exposure to metals, MT gene transfection, and MT gene knock-in techniques. Accordingly, in the present study, MT null astrocytes...
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| creator | Yao, Chang Ping Allen, Jeffrey W. Mutkus, Lysette A. Xu, Shao Bin Tan, Kim H. Aschner, Michael |
| description | The mechanisms associated with metallothionein (MT) gene regulation are complex and poorly understood. Only a modest increase in brain MT expression levels is attained by exposure to metals, MT gene transfection, and MT gene knock-in techniques. Accordingly, in the present study, MT null astrocytes isolated from transgenic mice deficient in MT-I and MT-II genes were introduced as a zero background model of MT expression. MT protein levels were determined by western blot analysis. MT proteins in MT-I and MT-II null astrocytes were undetectable. Transient MT-I gene transfection increased the levels of foreign MT expression in MT-I and MT-II null astrocytes by 2.3-fold above basal levels in wild-type astrocytes. Intracellular
Na
2
51
CrO
4
efflux and
d-[2,3-
3
H
]aspartate uptake were studied as indices of acute methylmercury (MeHg) (5 μM) cytotoxicity. In MT-I and MT-II knockout astrocytes MeHg led to significant (
p |
| doi_str_mv | 10.1016/S0006-8993(99)02211-8 |
| format | Article |
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Na
2
51
CrO
4
efflux and
d-[2,3-
3
H
]aspartate uptake were studied as indices of acute methylmercury (MeHg) (5 μM) cytotoxicity. In MT-I and MT-II knockout astrocytes MeHg led to significant (
p<0.01) increase in
Na
2
51
CrO
4
efflux and a significant (
p<0.05) decrease in the initial rate (1 min) of
d-[2,3-
3
H
]aspartate uptake compared to MT-I and MT-II knockout controls. Transfection of the MT-I gene in MT-I and MT-II null mice significantly (
p<0.01) decreased the effect of MeHg on
Na
2
51
CrO
4
efflux in MT null, as well as wild-type astrocytes. MT-I gene transfection in MT-I and MT-II null astrocytes reversed the inhibitory effect of MeHg on
d-[2,3-
3
H
]aspartate uptake, such that initial rates of uptake in MT-I transfected cells in the presence and absence of MeHg (5 μM) were indistinguishable. These results demonstrate that: (1) astrocytes lacking MTs are more sensitive to MeHg than those with basal MT protein levels, (2) the MT-I gene can be overexpressed in MT-I and MT-II null astrocytes by transient MT-I gene transfection, and (3) that foreign MT expression endows astrocytes with increased resistance to MeHg.</description><identifier>ISSN: 0006-8993</identifier><identifier>EISSN: 1872-6240</identifier><identifier>DOI: 10.1016/S0006-8993(99)02211-8</identifier><identifier>PMID: 10650127</identifier><identifier>CODEN: BRREAP</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Animals ; Aspartic Acid - pharmacokinetics ; Astrocytes - enzymology ; Biological and medical sciences ; Biological Transport - drug effects ; Cell Survival - drug effects ; Chemical and industrial products toxicology. Toxic occupational diseases ; Chromates ; Chromium Radioisotopes ; Cytotoxins - pharmacology ; Female ; Gene Expression Regulation, Enzymologic ; Knock-in ; Knockout ; Male ; Medical sciences ; Metallothionein ; Metallothionein - genetics ; Metallothionein - metabolism ; metallothionein I ; Metals and various inorganic compounds ; Methylmercury ; Methylmercury Compounds - toxicity ; Mice ; Mice, Knockout ; MT-I and MT-II null astrocytes ; Plasmids ; Sodium Compounds ; Toxicology ; Transfection ; Transgenic mouse</subject><ispartof>Brain research, 2000-02, Vol.855 (1), p.32-38</ispartof><rights>2000 Elsevier Science B.V.</rights><rights>2000 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c487t-cbeb71901de477a6a905e4067f0137d22b9eaa316f29f890e9712051a4ff75a63</citedby><cites>FETCH-LOGICAL-c487t-cbeb71901de477a6a905e4067f0137d22b9eaa316f29f890e9712051a4ff75a63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0006899399022118$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1264807$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10650127$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yao, Chang Ping</creatorcontrib><creatorcontrib>Allen, Jeffrey W.</creatorcontrib><creatorcontrib>Mutkus, Lysette A.</creatorcontrib><creatorcontrib>Xu, Shao Bin</creatorcontrib><creatorcontrib>Tan, Kim H.</creatorcontrib><creatorcontrib>Aschner, Michael</creatorcontrib><title>Foreign metallothionein-I expression by transient transfection in MT-I and MT-II null astrocytes confers increased protection against acute methylmercury cytotoxicity</title><title>Brain research</title><addtitle>Brain Res</addtitle><description>The mechanisms associated with metallothionein (MT) gene regulation are complex and poorly understood. Only a modest increase in brain MT expression levels is attained by exposure to metals, MT gene transfection, and MT gene knock-in techniques. Accordingly, in the present study, MT null astrocytes isolated from transgenic mice deficient in MT-I and MT-II genes were introduced as a zero background model of MT expression. MT protein levels were determined by western blot analysis. MT proteins in MT-I and MT-II null astrocytes were undetectable. Transient MT-I gene transfection increased the levels of foreign MT expression in MT-I and MT-II null astrocytes by 2.3-fold above basal levels in wild-type astrocytes. Intracellular
Na
2
51
CrO
4
efflux and
d-[2,3-
3
H
]aspartate uptake were studied as indices of acute methylmercury (MeHg) (5 μM) cytotoxicity. In MT-I and MT-II knockout astrocytes MeHg led to significant (
p<0.01) increase in
Na
2
51
CrO
4
efflux and a significant (
p<0.05) decrease in the initial rate (1 min) of
d-[2,3-
3
H
]aspartate uptake compared to MT-I and MT-II knockout controls. Transfection of the MT-I gene in MT-I and MT-II null mice significantly (
p<0.01) decreased the effect of MeHg on
Na
2
51
CrO
4
efflux in MT null, as well as wild-type astrocytes. MT-I gene transfection in MT-I and MT-II null astrocytes reversed the inhibitory effect of MeHg on
d-[2,3-
3
H
]aspartate uptake, such that initial rates of uptake in MT-I transfected cells in the presence and absence of MeHg (5 μM) were indistinguishable. These results demonstrate that: (1) astrocytes lacking MTs are more sensitive to MeHg than those with basal MT protein levels, (2) the MT-I gene can be overexpressed in MT-I and MT-II null astrocytes by transient MT-I gene transfection, and (3) that foreign MT expression endows astrocytes with increased resistance to MeHg.</description><subject>Animals</subject><subject>Aspartic Acid - pharmacokinetics</subject><subject>Astrocytes - enzymology</subject><subject>Biological and medical sciences</subject><subject>Biological Transport - drug effects</subject><subject>Cell Survival - drug effects</subject><subject>Chemical and industrial products toxicology. Toxic occupational diseases</subject><subject>Chromates</subject><subject>Chromium Radioisotopes</subject><subject>Cytotoxins - pharmacology</subject><subject>Female</subject><subject>Gene Expression Regulation, Enzymologic</subject><subject>Knock-in</subject><subject>Knockout</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Metallothionein</subject><subject>Metallothionein - genetics</subject><subject>Metallothionein - metabolism</subject><subject>metallothionein I</subject><subject>Metals and various inorganic compounds</subject><subject>Methylmercury</subject><subject>Methylmercury Compounds - toxicity</subject><subject>Mice</subject><subject>Mice, Knockout</subject><subject>MT-I and MT-II null astrocytes</subject><subject>Plasmids</subject><subject>Sodium Compounds</subject><subject>Toxicology</subject><subject>Transfection</subject><subject>Transgenic mouse</subject><issn>0006-8993</issn><issn>1872-6240</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUGP1SAUhYnROM_Rn6BhYYwuqkBbKKuJmTj6kjEuHNeE0ssMpoUnUDP9Q_5O6euLunPFveE7cHIOQs8peUsJ5e--EkJ41UlZv5byDWGM0qp7gHa0E6zirCEP0e4PcoaepPS9rHUtyWN0RglvCWVih35dhQju1uMJsh7HkO9c8OB8tcdwf4iQUtlxv-ActU8OfN4mCyavN87jzzcF1n44Dnvs53HEOuUYzJIhYRO8hZgKaSLoBAM-xJBPcn2rnU8ZazNnWD3cLeME0cxxwUUecrh3xuXlKXpk9Zjg2ek8R9-uPtxcfqquv3zcX76_rkzTiVyZHnpBJaEDNEJoriVpoSFcWEJrMTDWS9C6ptwyaTtJQArKSEt1Y61oNa_P0avt3eLxxwwpq8klA-OoPYQ5KSqamhNBCthuoIkhpQhWHaKbdFwUJWotSB0LUmv6Skp1LEh1Rffi9MHcTzD8o9oaKcDLE6CT0aMtYRuX_nKMNx1ZsYsNg5LGTwdRJVPaMTC4WLJVQ3D_cfIbT-iw_g</recordid><startdate>20000207</startdate><enddate>20000207</enddate><creator>Yao, Chang Ping</creator><creator>Allen, Jeffrey W.</creator><creator>Mutkus, Lysette A.</creator><creator>Xu, Shao Bin</creator><creator>Tan, Kim H.</creator><creator>Aschner, Michael</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>20000207</creationdate><title>Foreign metallothionein-I expression by transient transfection in MT-I and MT-II null astrocytes confers increased protection against acute methylmercury cytotoxicity</title><author>Yao, Chang Ping ; Allen, Jeffrey W. ; Mutkus, Lysette A. ; Xu, Shao Bin ; Tan, Kim H. ; Aschner, Michael</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c487t-cbeb71901de477a6a905e4067f0137d22b9eaa316f29f890e9712051a4ff75a63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Animals</topic><topic>Aspartic Acid - pharmacokinetics</topic><topic>Astrocytes - enzymology</topic><topic>Biological and medical sciences</topic><topic>Biological Transport - drug effects</topic><topic>Cell Survival - drug effects</topic><topic>Chemical and industrial products toxicology. Toxic occupational diseases</topic><topic>Chromates</topic><topic>Chromium Radioisotopes</topic><topic>Cytotoxins - pharmacology</topic><topic>Female</topic><topic>Gene Expression Regulation, Enzymologic</topic><topic>Knock-in</topic><topic>Knockout</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Metallothionein</topic><topic>Metallothionein - genetics</topic><topic>Metallothionein - metabolism</topic><topic>metallothionein I</topic><topic>Metals and various inorganic compounds</topic><topic>Methylmercury</topic><topic>Methylmercury Compounds - toxicity</topic><topic>Mice</topic><topic>Mice, Knockout</topic><topic>MT-I and MT-II null astrocytes</topic><topic>Plasmids</topic><topic>Sodium Compounds</topic><topic>Toxicology</topic><topic>Transfection</topic><topic>Transgenic mouse</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yao, Chang Ping</creatorcontrib><creatorcontrib>Allen, Jeffrey W.</creatorcontrib><creatorcontrib>Mutkus, Lysette A.</creatorcontrib><creatorcontrib>Xu, Shao Bin</creatorcontrib><creatorcontrib>Tan, Kim H.</creatorcontrib><creatorcontrib>Aschner, Michael</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>Yao, Chang Ping</au><au>Allen, Jeffrey W.</au><au>Mutkus, Lysette A.</au><au>Xu, Shao Bin</au><au>Tan, Kim H.</au><au>Aschner, Michael</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Foreign metallothionein-I expression by transient transfection in MT-I and MT-II null astrocytes confers increased protection against acute methylmercury cytotoxicity</atitle><jtitle>Brain research</jtitle><addtitle>Brain Res</addtitle><date>2000-02-07</date><risdate>2000</risdate><volume>855</volume><issue>1</issue><spage>32</spage><epage>38</epage><pages>32-38</pages><issn>0006-8993</issn><eissn>1872-6240</eissn><coden>BRREAP</coden><abstract>The mechanisms associated with metallothionein (MT) gene regulation are complex and poorly understood. Only a modest increase in brain MT expression levels is attained by exposure to metals, MT gene transfection, and MT gene knock-in techniques. Accordingly, in the present study, MT null astrocytes isolated from transgenic mice deficient in MT-I and MT-II genes were introduced as a zero background model of MT expression. MT protein levels were determined by western blot analysis. MT proteins in MT-I and MT-II null astrocytes were undetectable. Transient MT-I gene transfection increased the levels of foreign MT expression in MT-I and MT-II null astrocytes by 2.3-fold above basal levels in wild-type astrocytes. Intracellular
Na
2
51
CrO
4
efflux and
d-[2,3-
3
H
]aspartate uptake were studied as indices of acute methylmercury (MeHg) (5 μM) cytotoxicity. In MT-I and MT-II knockout astrocytes MeHg led to significant (
p<0.01) increase in
Na
2
51
CrO
4
efflux and a significant (
p<0.05) decrease in the initial rate (1 min) of
d-[2,3-
3
H
]aspartate uptake compared to MT-I and MT-II knockout controls. Transfection of the MT-I gene in MT-I and MT-II null mice significantly (
p<0.01) decreased the effect of MeHg on
Na
2
51
CrO
4
efflux in MT null, as well as wild-type astrocytes. MT-I gene transfection in MT-I and MT-II null astrocytes reversed the inhibitory effect of MeHg on
d-[2,3-
3
H
]aspartate uptake, such that initial rates of uptake in MT-I transfected cells in the presence and absence of MeHg (5 μM) were indistinguishable. These results demonstrate that: (1) astrocytes lacking MTs are more sensitive to MeHg than those with basal MT protein levels, (2) the MT-I gene can be overexpressed in MT-I and MT-II null astrocytes by transient MT-I gene transfection, and (3) that foreign MT expression endows astrocytes with increased resistance to MeHg.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>10650127</pmid><doi>10.1016/S0006-8993(99)02211-8</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Brain research, 2000-02, Vol.855 (1), p.32-38 |
| issn | 0006-8993 1872-6240 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Aspartic Acid - pharmacokinetics Astrocytes - enzymology Biological and medical sciences Biological Transport - drug effects Cell Survival - drug effects Chemical and industrial products toxicology. Toxic occupational diseases Chromates Chromium Radioisotopes Cytotoxins - pharmacology Female Gene Expression Regulation, Enzymologic Knock-in Knockout Male Medical sciences Metallothionein Metallothionein - genetics Metallothionein - metabolism metallothionein I Metals and various inorganic compounds Methylmercury Methylmercury Compounds - toxicity Mice Mice, Knockout MT-I and MT-II null astrocytes Plasmids Sodium Compounds Toxicology Transfection Transgenic mouse |
| title | Foreign metallothionein-I expression by transient transfection in MT-I and MT-II null astrocytes confers increased protection against acute methylmercury cytotoxicity |
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