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  • Title: Transgenic mouse blastocysts that overexpress metallothionein-I resist cadmium toxicity in vitro.
    Author: Lee DK, Fu K, Liang L, Dalton T, Palmiter RD, Andrews GK.
    Journal: Mol Reprod Dev; 1996 Feb; 43(2):158-66. PubMed ID: 8824913.
    Abstract:
    The role of metallothionein with regard to cadmium toxicity in vitro was investigated using preimplantation mouse blastocysts derived from a transgenic strain that constitutively overexpresses metallothionein-I transgenes (MT-I*). Northern blot and in situ hybridization revealed high levels of MT-I mRNA in transgenic blastocysts when compared with control blastocysts, and reverse transcriptase-polymerase chain reaction-amplified MT-I mRNA was almost exclusively MT-I*. Moreover, pulse-labeling experiments showed that the relative rate of synthesis of MT was 9-fold higher in transgenic blastocysts. Cadmium (Cd2+) toxicity was assessed after incubating blastocysts for 4 hr in Whitten's medium containing 50 microM Cd2+. Embryos that displayed abnormal morphology were judged "sensitive". Transgenic blastocysts were more resistant to cadmium-induced morphological changes than were control blastocysts. "Sensitive" and "resistant" blastocysts were individually genotyped by polymerase chain reaction, or they were transferred to foster mothers, and embryonic development to midterm was monitored. Of the blastocysts derived from mating heterozygous transgenic males with control females, 56% were transgenic before incubation with Cd2+, whereas 95% of the blastocysts that retained normal morphology after incubation were transgenic. Moreover, after Cd2+ exposure, transgenic blastocysts with normal morphology were nine times more likely to develop to midterm than were control blastocysts with normal morphology. Blastocysts with abnormal morphology failed to develop to midterm. These studies indicate that MT plays a central role in protection from Cd2+ toxicity within the physiological context of the developing mouse embryo.
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