Abstract
The polymerase chain reaction (PCR) differential display method is a powerful tool to detect and characterize alteration of gene expression in eukaryotic cells. In order to screen the differentially expressed genes between the adult (3 mo) and aged (24 mo) rats, the PCR differential display method was adopted in the present study. One differentially expressed cDNA band (C7-1) was identified and the aged rats expressed more the C7-1 gene than the adult rats in the frontal cortex, but not in the hippocampus and hypothalamus. The C7-1 cDNA band was recovered, reamplified, and subcloned as a probe in Northern blot analysis. A transcript of approx 2.8 kb was expressed in the frontal cortex of both the adult and aged rats, but the C7-1 mRNA level was increased for 52% in the aged rats. The C7-1 gene was then sequenced that contains 243 bp. We have found that the C7-1 cDNA shows no significant homology to any published genes, suggesting that the C7-1 gene is an unknown gene associated with aging. This study provides the first evidence to show that there is alteration in gene expression associated with aging by using the PCR differential display method.
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Azhar S., Cao L., and Reaven E. (1995) Alteration of the adrenal antioxidant defense system during aging in rats.J. Clin. Invest. 96, 1414–1424.
Chomczynski P. and Sacchi N. (1987) Single-step method of isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.Anal. Biochem. 162, 156–159.
Douglass J., McKinzie A. A., and Couceyro P. (1995) PCR differential display identifies a rat brain mRNA that is transcriptionally regulated by cocaine and amphetamine.J. Neurosci. 15, 2471–2481.
Finch C. (1990)Longevity, Senescence, and the Genome, University of Chicago Press, Chicago.
Gou J. P., Eyer J., and Leterrier J. F. (1995) Progressive hyperphosphorylation of neurofilament heavy subunits with aging.Biochem. Biophys. Res. Commun. 215, 368–376.
Gresik E. W., Wenk-Salamore K., Onetti-Muda A., Gubits R. M., and Shaw P. A. (1986) Effect of advanced age on the induction by androgen or thyroid hormone of epidermal growth factor and epidermal growth factor mRNA in the submandibular glands of C57BL/6 male mice.Mech. Aging Dev. 34, 175–189.
Huang A. M. and Lee E. H. Y. (1995) Analysis of gene expression associated with memory consolidation in rats using the differential display method.Soc. Neurosci. Abst. 21, 1933.
Imaizumi K., Tsuda M., Wanaka A., Tohyama M., and Takagi T. (1994) Differential expression of sgk mRNA, a member of the Ser/Thr protein kinase gene family, in rat brain after CNS injury.Mol. Brain Res. 26, 189–196.
Kanungo M. S. (1994) Changes in gene expression during aging, inGenes and Aging, Cambridge University Press, Cambridge, UK, pp. 167–242.
Kennedy B. K., Austriaco N. R., Zhang J., and Guarente L. (1995) Mutation in the silencing gene SIR4 can delay aging inS. cerevisiae.Cell 80, 485–496.
Liang P. and Pardee A. B. (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction.Science 257, 967–971.
Liang P., Averboukh L., and Pardee A. B. (1993) Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization.Nucleic Acids Res. 21, 3269–3275.
Milman N., Graudal N., and Andersen H. C. (1988) Acute phase reactants in the elderly.Clin. Chim. Acta. 176, 59–62.
Parhad I. M., Scott J. N., Cellars L. A., Bains J. S., Krekoski C. A., and Clark A. W. (1995) Axonal atrophy in aging is associated with a decline in neurofilament gene expression.J. Neurosci. Res. 41, 355–366.
Roy A. K., Nath T. S., Motwani N. M., and Chatterjee B. (1983) Age-dependent regulation of the polymorphic forms of alpha 2μ-globulin.J. Biol. Chem. 258, 10,123–10,127.
Semsei I., Rao G., and Richardson A. (1989) Changes in the expression of superoxide dismutase and catalse as a function of age and dietary restriction.Biochem. Biophys. Res. Commun. 164, 620–625.
Sierra F., Fey G. H., and Guigoz Y. (1989) T-kininogen gene expression is induced during aging.Mol. Cell Biol. 9, 5610–5616.
Whittemore S. R., Ebendal T., Larkfors L., Olson L., Seiger A., Stromberg I., and Persson H. (1986) Development and regional expression of beta nerve growth factor messenger RNA and protein in the rat central nervous system.Proc. Natl. Acad. Sci. USA 83, 817–821.
Yu C. E., Oshima J., Fu Y. H., Wijsman E. M., Hisama F., Alisch R. Matthews S., Nakura J., Miki T., Ouais S., Martin G. M., Mulligan J., and Schellenberg G. D. (1996) Positional cloning of the Wernerís syndrome gene.Science 272, 258–262.
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Wu, H.C., Lee, E.H.Y. Identification of a rat brain gene associated with aging by PCR differential display method. J Mol Neurosci 8, 13–18 (1997). https://doi.org/10.1007/BF02736859
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DOI: https://doi.org/10.1007/BF02736859