Membrane glycopeptides from old and young human erythrocytes.
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ABSTRACT:
SUBMITTER: Balduini C
PROVIDER: S-EPMC1168036 | biostudies-other | 1974 Jun
REPOSITORIES: biostudies-other
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Project description:Understanding gene expression changes over the lifespan of cells is of fundamental interest and gives important insights into processes related to maturation and ageing. This study was undertaken to understand the global transcriptome changes associated with ageing in fish erythrocytes. Fish erythrocytes retain their nuclei throughout their lifetime and they are transcriptionally and translationally active. However, they lose important functions during their lifespan in the circulation. We separated rainbow trout (Oncorhynchus mykiss) erythrocytes into young and old fractions using fixed angle-centrifugation and analyzed transcriptome changes using RNA sequencing (RNA-seq) technology and quantitative real-time PCR. We found 930 differentially expressed between young and old erythrocyte fractions; 889 of these showed higher transcript levels in young, while only 34 protein-coding genes had higher transcript levels in old erythrocytes. In particular ion binding, signal transduction, membrane transport, and various enzyme classes are affected in old erythrocytes. The transcripts with higher levels in old erythrocytes were associated with 7 different GO terms within biological processes and 9 within molecular functions and cellular components respectively. Our study furthermore found several highly abundant transcripts as well as a number of differentially expressed genes for which the protein products are currently not known revealing the gaps of knowledge in most non-mammalian vertebrates. Our data provide the first insight into changes involved in ageing on the transcriptional level and thus opens new perspectives for the study of maturation processes in fish erythrocytes.
2018-02-13 | GSE106570 | GEO
Project description:To gain knowledge about the behaviour of calpastatin (the specific inhibitor of the Ca(2+)-dependent thiol protease calpain) in the intact cell, we analysed the inhibitor by specific antibodies and determined its activity in erythrocytes from individuals 20-34 years old (young) and 70-93 years old (old). Differences between old and young in the behaviour of erythrocyte calpastatin were observed. Erythrocytes of old individuals had lower amounts of calpastatin and less calpastatin activity than those of young ones. A difference between old and young was also found in the molecular-mass distribution of calpastatin subunits. Increasing the erythrocyte Ca2+ induced changes in calpastatin in young individuals, rendering it similar to calpastatin in cells of old individuals. When calpastatin (isolated from erythrocytes of a young individual) was added to erythrocyte membranes, the initial binding and subsequent association of calpastatin with the membrane were lower in old than in young individuals. We had previously found that calpain binding and activation were enhanced in erythrocyte membranes from old individuals, along with enhanced degradation of band 3 (a major erythrocyte transmembrane anion-transport protein). The overall results indicate an interaction of calpain with calpastatin in the intact cell. Enhanced activation of erythrocyte calpain and degradation of calpastatin occur under conditions of increased cellular Ca2+ and in cells of the aged.
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