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Altered expression of Apoptotic Machinery genes explains the decrease of oocytes competence with aging. MR Guglielmino 1 , M Vento 2 , M Santonocito 1 , P Borzì 2 , M Ragusa 1 , D Barbagallo 1 , I Casciano 3 , L Statello 1 , P Scollo 2 , M Romani 3 , M Purrello 1 , C Di Pietro 1 . 1 Dipartimento Gian Filippo Ingrassia, Sezione di Biologia, Genetica, Genomica Cellulare e Molecolare Giovanni Sichel, Università degli Studi di Catania; E-mail: dipietro@unict.it 2 Servizio di PMA/Azienda Ospedaliera Cannizzaro, Catania; E-mail: mrln.vento@gmail.com 3 Tumor Genetics and Epigenetics, Istituto Nazionale per la Ricerca sul Cancro - IST Genova; E-mail: massimo.romani@istge.it To identify the biomolecular bases of reduced oocyte competence with aging, we compared the Apoptotic Machinery (AM) transcriptome of pooled MII oocytes from two cohorts of women, respectively younger than 35 (young) and older than 38 years (old). We found that the expression profile of AM genes is strongly dependent on maternal age. In fact, 36 out of the 84 AM genes analyzed showed a specific age-dependent expression profile: 16 proapoptotic genes were considerably upregulated and 7 antiapoptotic genes were downregulated in oocytes from older women. Through Real-Time PCR on single MII oocytes (6 from young and 5 from older women), we confirmed that the mRNAs encoding prosurvival proteins Bcl2 and cFLAR are more abundant in oocytes from younger women (p
Towards understanding of life and death regulation at CD95/Fas Nicolai Fricker(1,3), Carina Pforr(1), Roland Eils(2,3,4), Peter H. Krammer(1), Inna N. Lavrik(1,3) 1Division of Immunogenetics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany 2Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany 3Bioquant, University of Heidelberg, 69120 Heidelberg, Germany 4Department for Bioinformatics and Functional Genomics, Institute for Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany CD95 (APO-1/Fas) is a member of the death receptor family. CD95 stimulation leads to the induction of apoptotic as well as non-apoptotic pathways. CD95 signaling starts with the formation of the death-inducing signaling complex (DISC). The DISC consists of CD95, the adaptor molecule FADD (Fas-Associated Death Domain), procaspase-8, procaspase-10 and c- FLIPL/S/R (the cellular FLICE-inhibitory proteins). The regulation of life and death decisions at the DISC is largely influenced by the ratio between pro- and anti-apoptotic DED (death effector domain)–containing proteins and is poorly understood. We have applied a systems biology approach to understand the regulation of life/death decisions in CD95 signaling pathway in quantitative terms. In our experiments we have used HeLa cells overexpressing various components of the CD95 system. Stimulation of HeLa cells has led to the induction of both apoptotic and non-apoptotic pathways depending on the concentration of the CD95 DISC components. We have quantitatively analyzed apoptotic and non-apoptotic responses applying single-cell and population-based measurements. Based on these experimental data a mathematical model of the CD95 DISC has been constructed. The model takes into account the complex interplay between various DED-containing proteins of the DISC, e.g. procaspase-8 and c-FLIP isoforms as well as their cleavage products. The application of systems biology allowed to identify new mechanisms of regulation of CD95 signaling on the quantitative level and has demonstrated how exact concentrations of c-FLIP and procaspase-8 and their cleavage products at the DISC can determine life/death decisions. 23
Page 1 and 2: Luxembourg, January 26th to 28th, 2
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Session 9: Neurodegenerative Diseas
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List of participants (In alphabetic
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Mrs. Emilie Bana Laboratoire d'Ing
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Dr. Alessandro Corsaro Laboratory o
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Pr. Paul Fisher Microbial Cell Biol
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Mr. Kasper Hoebe Cincinnati Childre
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Mr. Hendrik Koopmans Cell Biology W
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Mr. Romuald Menth Technical Support
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We thank our sponsors:
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