Cadmium chloride (CdCl2)-induced metallothionein (MT) expression in neonatal rat primary astrocyte cultures

L. Rising, D. Vitarella, H. K. Kimelberg, Michael Aschner

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Metallothionein (MT) protein and mRNA levels were studied following exposure of rat neonatal primary astrocyte cultures to cadmium chloride (CdCl2). MT mRNA was probed on Northern blots with a32P labeled synthetic cDNA probe specific for rat MT mRNA. The probe hybridizes to a single mRNA with a size appropriate for MT, approximately 550 bases. Expression of MT-I mRNA in astrocyte monolayers exposed to 2 × 10-6 M CdCl2 for 6 h was increased approximately 5-fold (9.7 fg/μg total RNA) over MT-I mRNA levels in controls (2 fg/μg total RNA). MT-I mRNA could also be detected in untreated cells, suggesting constitutive MT expression in these cells. Western-blot analysis revealed a marked increase in MT protein levels upon exposure to CdCl2 (1 × 10-6 M; 96 h). Consistent with the constitutive expression of MTs both at the mRNA level and protein level, we have also demonstrated a time-dependent increase in MT-immunoreactivity in astrocytes exposed to CdCl2. The present study suggests that astrocytes constitutively express MTs, and that MT-induction by CdCl2 may be an example of a generalized increase in MTs in response to heavy metal exposure, thus protecting astrocytes, and perhaps also indirectly, juxtaposed neurons from the neurotoxic effects of heavy metals.

Original languageEnglish (US)
Pages (from-to)91-98
Number of pages8
JournalBrain Research
Volume678
Issue number1-2
DOIs
StatePublished - Apr 24 1995
Externally publishedYes

Fingerprint

Cadmium Chloride
Metallothionein
Astrocytes
Messenger RNA
Heavy Metals
RNA
Proteins
Northern Blotting
Complementary DNA
Western Blotting

Keywords

  • Astrocyte
  • Cadmium chloride
  • d-Aspartate
  • Metallothionein
  • Rat

ASJC Scopus subject areas

  • Developmental Biology
  • Molecular Biology
  • Clinical Neurology
  • Neuroscience(all)

Cite this

Cadmium chloride (CdCl2)-induced metallothionein (MT) expression in neonatal rat primary astrocyte cultures. / Rising, L.; Vitarella, D.; Kimelberg, H. K.; Aschner, Michael.

In: Brain Research, Vol. 678, No. 1-2, 24.04.1995, p. 91-98.

Research output: Contribution to journalArticle

@article{4e1f8447fcc1495db0217ab94d5ea22f,
title = "Cadmium chloride (CdCl2)-induced metallothionein (MT) expression in neonatal rat primary astrocyte cultures",
abstract = "Metallothionein (MT) protein and mRNA levels were studied following exposure of rat neonatal primary astrocyte cultures to cadmium chloride (CdCl2). MT mRNA was probed on Northern blots with a32P labeled synthetic cDNA probe specific for rat MT mRNA. The probe hybridizes to a single mRNA with a size appropriate for MT, approximately 550 bases. Expression of MT-I mRNA in astrocyte monolayers exposed to 2 × 10-6 M CdCl2 for 6 h was increased approximately 5-fold (9.7 fg/μg total RNA) over MT-I mRNA levels in controls (2 fg/μg total RNA). MT-I mRNA could also be detected in untreated cells, suggesting constitutive MT expression in these cells. Western-blot analysis revealed a marked increase in MT protein levels upon exposure to CdCl2 (1 × 10-6 M; 96 h). Consistent with the constitutive expression of MTs both at the mRNA level and protein level, we have also demonstrated a time-dependent increase in MT-immunoreactivity in astrocytes exposed to CdCl2. The present study suggests that astrocytes constitutively express MTs, and that MT-induction by CdCl2 may be an example of a generalized increase in MTs in response to heavy metal exposure, thus protecting astrocytes, and perhaps also indirectly, juxtaposed neurons from the neurotoxic effects of heavy metals.",
keywords = "Astrocyte, Cadmium chloride, d-Aspartate, Metallothionein, Rat",
author = "L. Rising and D. Vitarella and Kimelberg, {H. K.} and Michael Aschner",
year = "1995",
month = "4",
day = "24",
doi = "10.1016/0006-8993(95)00170-U",
language = "English (US)",
volume = "678",
pages = "91--98",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Cadmium chloride (CdCl2)-induced metallothionein (MT) expression in neonatal rat primary astrocyte cultures

AU - Rising, L.

AU - Vitarella, D.

AU - Kimelberg, H. K.

AU - Aschner, Michael

PY - 1995/4/24

Y1 - 1995/4/24

N2 - Metallothionein (MT) protein and mRNA levels were studied following exposure of rat neonatal primary astrocyte cultures to cadmium chloride (CdCl2). MT mRNA was probed on Northern blots with a32P labeled synthetic cDNA probe specific for rat MT mRNA. The probe hybridizes to a single mRNA with a size appropriate for MT, approximately 550 bases. Expression of MT-I mRNA in astrocyte monolayers exposed to 2 × 10-6 M CdCl2 for 6 h was increased approximately 5-fold (9.7 fg/μg total RNA) over MT-I mRNA levels in controls (2 fg/μg total RNA). MT-I mRNA could also be detected in untreated cells, suggesting constitutive MT expression in these cells. Western-blot analysis revealed a marked increase in MT protein levels upon exposure to CdCl2 (1 × 10-6 M; 96 h). Consistent with the constitutive expression of MTs both at the mRNA level and protein level, we have also demonstrated a time-dependent increase in MT-immunoreactivity in astrocytes exposed to CdCl2. The present study suggests that astrocytes constitutively express MTs, and that MT-induction by CdCl2 may be an example of a generalized increase in MTs in response to heavy metal exposure, thus protecting astrocytes, and perhaps also indirectly, juxtaposed neurons from the neurotoxic effects of heavy metals.

AB - Metallothionein (MT) protein and mRNA levels were studied following exposure of rat neonatal primary astrocyte cultures to cadmium chloride (CdCl2). MT mRNA was probed on Northern blots with a32P labeled synthetic cDNA probe specific for rat MT mRNA. The probe hybridizes to a single mRNA with a size appropriate for MT, approximately 550 bases. Expression of MT-I mRNA in astrocyte monolayers exposed to 2 × 10-6 M CdCl2 for 6 h was increased approximately 5-fold (9.7 fg/μg total RNA) over MT-I mRNA levels in controls (2 fg/μg total RNA). MT-I mRNA could also be detected in untreated cells, suggesting constitutive MT expression in these cells. Western-blot analysis revealed a marked increase in MT protein levels upon exposure to CdCl2 (1 × 10-6 M; 96 h). Consistent with the constitutive expression of MTs both at the mRNA level and protein level, we have also demonstrated a time-dependent increase in MT-immunoreactivity in astrocytes exposed to CdCl2. The present study suggests that astrocytes constitutively express MTs, and that MT-induction by CdCl2 may be an example of a generalized increase in MTs in response to heavy metal exposure, thus protecting astrocytes, and perhaps also indirectly, juxtaposed neurons from the neurotoxic effects of heavy metals.

KW - Astrocyte

KW - Cadmium chloride

KW - d-Aspartate

KW - Metallothionein

KW - Rat

UR - http://www.scopus.com/inward/record.url?scp=0028988105&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028988105&partnerID=8YFLogxK

U2 - 10.1016/0006-8993(95)00170-U

DO - 10.1016/0006-8993(95)00170-U

M3 - Article

C2 - 7620903

AN - SCOPUS:0028988105

VL - 678

SP - 91

EP - 98

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1-2

ER -