Analysis of aging-associated genes in myocardium : application of RT-PCR differential display method.

这已经知道心脏的physiological function of the heart is altered according to the agingfor example,isoproterenol和endothelin-1 are decreased by aging in the rat positive inotropic effectsventricles. While the binding capacity for b-adrenergic receptor was decreasedthe specific binding capacity of endothelin-1 was increased in the aged rats. in sarcoplasmicreticulum, the activity of Ca D12+ D1 was reduced. In coronary arterial endothelium，nitric oxide production was very low-in aged rat. The molecular basis of these age-dependentfunctional changes not been understood. Differential gene expression provides the basis for manyfundamental cellular processes with differentiation，development and aging. The identification and cloning of genes whose expression is modulated is animportant step in advancing our understanding of aging phenomena. in order to isolate andcharacterize gene产品differentially expressed in aging， we applied the diffe…rential display reverse transcription-polymerase chain reaction for screening thosegenes in 2-3 months-old和27 months-old Fischer 344 rat ventricles. Total RNAs were isolated byacid-guanidium-phenol-chloroform method. Messenger RNAs were obtained from total RNA by using oligod(T) column. The first-strand cDNAs from each RNA were synthesized with three oligo-d(T)-anchorPCR was carried out with anchor primers and eight arbitrary primersand the products were analyzed on sequence gels. the 19 unique PCR products displayed increasedexpression in aged heart，66 clones decreased in aged heart. Differential expression was confirmed by Northern blotanalysis or reverse Northern blotting. The clones included genes for mitochondrial metabolism suchas cytochrome c oxidase, myosin heavy chain，cell surface glycoproteins and several kinases/phosphatases for signal transduction. 31 clones weresequenced and 14 clones unknown genes. The method used in this study is valuable for studyingthe age-associated gene expression我们持续分析未知的性别识别在这方面研究少

It has been known that the physiological function of the heart is altered according to the aging. For example, the positive inotropic effects of isoproterenol and endothelin-1 are decreased by aging in the rat ventricles. While the binding capacity for b-adrenergic receptor was decreased, the specific binding capacity of endothelin-1 was increased in the aged rats. In sarcoplasmic reticulum, the activity of CaィイD12+ィエD1 was reduced. In coronary arterial endothelium, nitric oxide production was very low-in aged rat. The molecular basis of these age-dependent functional changes has not been understood. Differential gene expression provides the basis for many fundamental cellular processes associated with differentiation, development and aging. The identification and cloning of genes whose expression is modulated is an important step in advancing our understanding of aging phenomena. In order to isolate and characterize gene products differentially expressed in aging, we applied the diffe … More rential display reverse transcription-polymerase chain reaction for screening those genes in 2-3 months-old and 27 months-old Fischer 344 rat ventricles. Total RNAs were isolated by acid-guanidium-phenol-chloroform method. Messenger RNAs were obtained from total RNA by using oligo d(T) column. The first-strand cDNAs from each RNA were synthesized with three oligo-d(T)-anchor primers. PCR was carried out with anchor primers and eight arbitrary primers, and the products were analyzed on sequence gels. The 19 unique PCR products displayed increased expression in aged heart, while 66 clones were decreased in aged heart. Differential expression was confirmed by Northern blot analysis or reverse Northern blotting. The clones included genes for mitochondrial metabolism such as cytochrome c oxidase, myosin heavy chain, cell surface glycoproteins and several kinases/phosphatases for signal transduction. 31 clones were sequenced and 14 clones were unknown genes. The method used in this study is valuable for studying the age-associated gene expression, and we are continuing to analyze the unknown genes identified in this study. Less