Fleming J E, Walton J K, Dubitsky R, Bensch K G
Ryoichi Sasakawa Center for Aging Research, Linus Pauling Institute of Science and Medicine, Palo Alto, CA 94306.
Proc Natl Acad Sci U S A. 1988 Jun;85(11):4099-103. doi: 10.1073/pnas.85.11.4099.
We used high-resolution two-dimensional polyacrylamide gel electrophoresis to evaluate the effect of aging on the heat shock response in Drosophila melanogaster. Although the aging process is not well understood at the molecular level, recent observations suggest that quantitative changes in gene expression occur as these fruit flies approach senescence. Such genetic alterations are in accord with our present data, which clearly show marked differences in the synthesis of heat shock proteins between young and old fruit flies. In 10-day-old flies, a heat shock of 20 min results in the expression of 14 new proteins as detectable by two-dimensional electrophoresis of [35S]methionine-labeled polypeptides, whereas identical treatment of 45-day-old flies leads to the expression of at least 50 new or highly up-regulated proteins. In addition, there is also a concomitant increase in the rate of synthesis of a number of the normal proteins in the older animals. Microdensitometric determinations of the low molecular weight heat shock polypeptides on autoradiographs of five age groups revealed that their maximum expression occurs at 47 days for a population of flies with a mean life span of 33.7 days. Moreover, a heat shock effect similar to that observed in senescent flies occurs in young flies fed canavanine, an arginine analogue, before heat shock.
我们使用高分辨率二维聚丙烯酰胺凝胶电泳来评估衰老对黑腹果蝇热休克反应的影响。尽管衰老过程在分子水平上尚未被充分理解,但最近的观察表明,随着这些果蝇接近衰老,基因表达会发生定量变化。这种基因改变与我们目前的数据一致,我们的数据清楚地显示了年轻果蝇和老年果蝇热休克蛋白合成的显著差异。在10日龄的果蝇中,20分钟的热休克导致14种新蛋白质的表达,这可通过对[35S]甲硫氨酸标记的多肽进行二维电泳检测到,而对45日龄果蝇进行相同处理则导致至少50种新的或高度上调的蛋白质表达。此外,老年动物中许多正常蛋白质的合成速率也随之增加。对五个年龄组的放射自显影片上低分子量热休克多肽的显微密度测定表明,对于平均寿命为33.7天的果蝇群体,它们的最大表达出现在47天。此外,在热休克前喂食刀豆氨酸(一种精氨酸类似物)的年轻果蝇中,会出现与衰老果蝇中观察到的类似的热休克效应。