Cadmium chloride (CdCl 2)-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 (CdCl 2). MT mRNA was probed on Northern blots with a 32P 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 CdCl 2 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 CdCl 2 (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 CdCl 2. The present study suggests that astrocytes constitutively express MTs, and that MT-induction by CdCl 2 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.