Effect of calorie restriction on the metabolic history of chronologically aging yeast

Mechanism of liponecrosis, a distinct mode of programmed cell death

“…Measurement of changes in the pH of culture medium, 72 measurement of PE level in the extracellular leaflet of the PM, 22 cellular respiration assay, 72 monitoring of the mitochondrial membrane potential, 72 ROS measurement, 73 preparation of cellular extracts and a microanalytic assay for measuring ATP, 74 recovery of insoluble aggregates of denatured proteins and an assay for monitoring the extent of protein aggregation, 73 immunodetection of carbonyl groups of oxidatively damaged cellular proteins, 73 SDS-PAGE, 75 and an assay for measuring oxidatively damaged cellular lipids 13 were performed as previously described.…”

Lithocholic bile acid accumulated in yeast mitochondria orchestrates a development of an anti-aging cellular pattern by causing age-related changes in cellular proteome

“…Preparation of total cell lysates, 37 purification of mitochondria, 81 SDS-PAGE, 82 quantitative mass spectrometric analysis of proteins 37 and statistical analysis 83 were performed as previously described.…”

“…Yeast cells for these experiments were cultured under caloric restriction (CR) conditions on 0.2% glucose rather than under non-CR conditions on 2% glucose. We chose CR conditions for the following reasons: (1) mitochondria, the organelles whose LCA-driven ability to operate as signaling compartments in yeast chronological aging we investigated in this study, are significantly more abundant and functionally active in yeast cells limited in calorie supply than they are in yeast cells on a high-calorie diet; 37,38 …”

Metabolomic and Lipidomic Analyses of Chronologically Aging Yeast

“…Recent metabolomic and lipidomic analyses of the metabolic history of chronologically aging yeast cells strongly suggest that their longevity is defined by a pattern of metabolism and organelle dynamics established prior to cell entry into a non-proliferative state in a genotype-, diet-, and pharmacological interventiondependent manner [8,9,[20][21][22][23][25][26][27][28]. This chapter describes detailed protocols for such high-throughput analyses that have been used to measure the levels of trehalose, glycogen, ethanol, and acetic acid as well as to quantitatively assess the entire complement of cellular lipids.…”

“…Due to the relatively short and easily monitored replicative and chronological life spans of this genetically and biochemically manipulable unicellular eukaryote with annotated genome, it has been successfully used to (1) identify numerous novel longevity genes, many of which have been later implicated in regulating longevity of multicellular eukaryotic organisms; (2) establish the chemical nature of molecular damage that causes cellular and organismal aging and accelerates the onset of age-related diseases; and (3) identify a number of longevity-extending small molecules, many of which have been later shown to slow down aging, improve health, attenuate age-related pathologies and delay the onset of age-related diseases in multicellular eukaryotes [3,5,8,9,[18][19][20][21][22][23][24][25][26][27][28].…”