Transfection and overexpression of metallothionein-I in neonatal rat primary astrocyte cultures and in astrocytoma cells increases their resistance to methylmercury-induced cytotoxicity

Chang Ping Yao, Jeffrey W. Allen, Dawn R. Conklin, Michael Aschner

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Metallothionein-I (MT-I) was expressed in neonatal rat primary astrocyte cultures and an astrocytoma cell line by pGFAP-MT-I plasmid transfection under the control of the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter. Following transient transfection of the pGFAP-MT-I plasmid, MT-I mRNA and MT-I protein levels were determined by northern blot and immunoprecipitation analyses, respectively. The ability of cells over- expressing MT-I to withstand acute methylmercury (MeHg) treatment was measured by the release of preloaded Na2 51CrO4, an indicator of membrane integrity. Transfection with the pGFAP-MT-I plasmid led to increased mRNA (2.5-fold in astrocytes and 7.4-fold in astrocytomas) and MT-I protein (2.4- fold in astrocytes and 4.0-fold in astrocytomas) levels compared with their respective controls. Increased expression of MT-I was associated with attenuated release of Na2 51CrO4 upon MeHg (5 μM) treatment. These results demonstrate that MT-I can be highly expressed both in primary astrocyte cultures and astrocytomas by pGFAP-MT-I plasmid transfection, and lend credence to the hypothesis that increased expression of MT-I affords protection against the cytotoxic effects of MeHg. Taken together, the data suggest that MT offer effective cellular adaptation to MeHg cytotoxicity.

Original languageEnglish (US)
Pages (from-to)414-420
Number of pages7
JournalBrain Research
Volume818
Issue number2
DOIs
StatePublished - Feb 13 1999
Externally publishedYes

Fingerprint

Metallothionein
Astrocytoma
Astrocytes
Transfection
Plasmids
Messenger RNA
Glial Fibrillary Acidic Protein
Immunoprecipitation
Northern Blotting
Proteins

Keywords

  • Astrocyte
  • Astrocytoma
  • Glial fibrillary acidic protein
  • Metallothionein-I
  • Methylmercury

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Transfection and overexpression of metallothionein-I in neonatal rat primary astrocyte cultures and in astrocytoma cells increases their resistance to methylmercury-induced cytotoxicity. / Yao, Chang Ping; Allen, Jeffrey W.; Conklin, Dawn R.; Aschner, Michael.

In: Brain Research, Vol. 818, No. 2, 13.02.1999, p. 414-420.

Research output: Contribution to journalArticle

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