Mapping Genetic Polymorphisms Affecting Natural Variation in Drosophila Longevity

Maria De Luca and Jeff Leips Summary Analyses of mutations affecting life span in model organisms have revealed a number of genes that regulate longevity in evolutionarily conserved signaling pathways. These studies suggest that genes involved in insulin-like signaling pathways, metabolism, stress response, and prevention of oxidative damage influence life span. However, we do not know whether functional polymorphisms at these candidate genes affect population variation in longevity. To...

Germinal Vesicle Nuclear Transfer

Collection of GV oocytes and IVM of denuded oocytes. The isolation and culture of immature oocytes are performed essentially following procedures described (24). Female mice are primed by a single i.p. injection of 5 IU of PMSG 44-46 h prior to collection of GV oocytes, and euthanized humanely by cervical dislocation (25). Abdominal areas are sterilized with 70 ethanol. Skin and peritoneum are cut to open and expose the abdominal cavity. Ovaries are removed and ovaries with antral follicles...

Identifying Genes that Extend Life Span Using a High Throughput Screening System

Cuiying Chen and Roland Contreras Summary We developed a high-throughput functional genomic screening system that allows identification of genes prolonging lifespan in the baker's yeast Saccharomyces cerevisiae. The method is based on isolating yeast mother cells with a higher than average number of cell divisions as indicated by the number of bud scars on their surface. Fluorescently labeled wheat germ agglutinin (WGA) was used for specific staining of chitin, a major component of bud scars....

References

W., Marchie, A., et al. 2003 Effects of a dietary portfolio of cholesterol-lowering foods vs lovastatin on serum lipids and C-reactive protein. see comment . JAMA 290, 502-510. 2. Jenkins, D. J., Kendall, C. W., Marchie, A., Augustin, L. S. 2004 Too much sugar, too much carbohydrate, or just too much comment . Am. J. Clin. Nutr. 79, 711-712. 3. Jenkins, D. J., Kendall, C. W., Marchie, A., et al. 2003 Type 2 diabetes and the vegetarian diet. Am. J. Clin. Nutr. 78 3...

Methods for Cell Sorting of Young and Senescent Cells

Passos and Thomas von Zglinicki Cellular senescence, the ultimate and irreversible loss of replicative capacity of cells in primary culture, has been a popular model for studying the aging process. However, the replicative life span of human fibroblasts is heterogeneous even in clonal populations, with the fraction of senescent cells increasing at each population doubling, rather than all cells entering senescence simultaneously. Thus, the study of individual cells in a mass culture is...

Preparation of Yeast for Replicative Life Span Analysis

Replicative Life Span

1. 2.2X supplemented yeast extract peptone YEP media 22.2 g L Bacto-yeast extract Becton, Dickinson and Company, Franklin, NJ , 44.4 g L Bacto-peptone Becton, Dickinson and Company, Franklin, NJ in water and filter sterilized with a 0.22-pm vacuum filter. Supplement the media by adding 40 mL L 0.5 adenine solution and 20 mL L for the remaining 1 amino acid solutions see Notes 1-3 . 2. 4 agar Bacto-agar Becton, Dickinson and Company, Franklin, NJ 8 g of agar is added into 200 mL of water and...