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Real-time fluorescent quantitative RT-PCR assay for the expression of metallothioneins in rat hippocampal neurons
Author(s): Hai-Hong Qin; Fu-Di Wang; Jun-Sheng Guo; Hui Shen; Run-Ping Li
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Paper Abstract

Metallothioneins (MTs) are short, cysteine-rich proteins for heavy metal homeostasis and detoxification; they can bind a variety of heavy metals and also act as radical scavengers. In brain cells, they play a neuroprotective role in many aspects. However, because the previous methods can't quantify their gene expression at the mRNA level, their regulation in brain, especially in neurons, is not well known by now. In this study, we use a more accurate method, the real-time fluorescent quantitative RT-PCR technique, to determine the expression of three MT isomers on 100 μM zinc exposure after 0, 2, 4, 6 and 8 hours in primary culture rat hippocampal neurons. The result shows that the expression of all three MT isomers was higher compared with the values determined by other methods. This means that the roles played by neuron MTs in protecting neurons injury on zinc fluctuation was even stronger than what has been suspected before. In conclusion, our study proved that the real-time fluorescent quantitative RT-PCR technique is a simple, rapid and more precise method than previous techniques in the detection of gene expression, especially for those genes with low abundant mRNA. Our study also suggest that may of the past studies about gene expression should be verified by real-time Fluorescent quantitative RT-PCR once more in order to reach a more scientific explanation on certain problem.

Paper Details

Date Published: 29 July 2004
PDF: 5 pages
Proc. SPIE 5486, ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics, (29 July 2004); doi: 10.1117/12.572446
Show Author Affiliations
Hai-Hong Qin, Second Military Medical Univ. (China)
Fu-Di Wang, Second Military Medical Univ. (China)
Jun-Sheng Guo, Second Military Medical Univ. (China)
Hui Shen, Second Military Medical Univ. (China)
Run-Ping Li, Second Military Medical Univ. (China)

Published in SPIE Proceedings Vol. 5486:
ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics
Ruikang K. Wang; Jeremy C. Hebden; Alexander V. Priezzhev; Valery V. Tuchin, Editor(s)

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