Effects of Aging and Calorie Restriction on the Global Gene Expression Profiles of Mouse Testis and Ovary. Mus musculus

AGEMAP is an initiative of the National Institute on Aging (NIA) designed to study the aging process on mice with Caloric Restriction (CR) versus Ad Libitum (AL) diet. The aim of the AGEMAP project is to provide a valuable source of information about genetic influences involved in aging of the major organ systems. A total of 67 mice (C57BL/6) were sacrificed. Both male and female mice were divided in 7 categories based on the diet and the age. Four categories in ad libitum (1 month, 6 month,16 month and 24 month) and three categories in caloric restriction (6 month, 16 month and 24 month). In the current study we focused on gonads, using microarray hybridization (Agilent platform 44K array) to identify gene expression changes age-associated CR-associated, between young versus old mice at different time points. Keywords: development or differentiation design,replicate design,stimulus or stress design,time series design Overall design: The experiment was done as time-course type. Four time points for mice on ad libitum diet (1 month, 6 month, 16 month and 24 month), and three time points on caloric restriction diet. For each time point we used two biological replicates. We analyzed gene expression patterns in ovaries and testes sampled from 1-, 6-, 16-, and 24-month-old C57BL/6 mice fed on AL and CR diets (Figure 1A and D). Mice were kept on an AL diet until 14 weeks of age and then split into AL and CR gonads . Two individual animals for each condition were used for the microarray studies. We examined a total of 28 samples (two AL ovaries at each of 1, 6, 16, and 24 months; two CR ovaries at 6, 16, and 24 months; two AL testes at 1, 6, 16, and 24 months; and two CR testes at 6, 16, and 24 months). These 28 samples were hybridized on whole-genome oligonucleotide microarrays bearing approximately 44,000 probes, representing 25,585 nonredundant genes with gene symbols (Additional file 1). The same RNA samples were used for quantitative reverse-transcription polymerase chain reaction (RT-PCR)

AGEMAP是美国国家衰老研究所（National Institute on Aging, NIA）发起的一项研究计划，旨在通过对比热量限制（Caloric Restriction, CR）与自由采食（Ad Libitum, AL）两种饮食方式，探究小鼠的衰老进程。AGEMAP项目的目标是为研究主要器官系统衰老过程中的遗传影响因素提供宝贵的信息资源。本次研究共处死67只C57BL/6品系小鼠，按饮食类型与月龄将雌雄小鼠分为7组：自由采食组设4个月龄节点（1、6、16、24月龄），热量限制组设3个月龄节点（6、16、24月龄）。本研究聚焦性腺组织，采用微阵列杂交（microarray hybridization）技术（安捷伦Agilent平台44K芯片），旨在识别不同时间节点下年轻与老年小鼠中与衰老、热量限制相关的基因表达变化。关键词：发育或分化设计、重复设计、刺激或应激设计、时间序列设计。总体实验设计：本实验为时间序列类研究。自由采食饮食组设置4个时间点，热量限制饮食组设置3个时间点，每个时间点均使用2个生物学重复。本研究分析了分别取自AL与CR饮食组的1、6、16、24月龄C57BL/6小鼠的卵巢与睾丸组织的基因表达模式（见图1A与图1D）。所有小鼠在14周龄前均采用AL饮食饲养，14周龄后被随机分为AL组与CR组并采集性腺组织。每个实验条件均使用2只独立个体进行微阵列检测。本次研究共制备28份检测样本，具体包括：1、6、16、24月龄AL组卵巢各2份；6、16、24月龄CR组卵巢各2份；1、6、16、24月龄AL组睾丸各2份；6、16、24月龄CR组睾丸各2份。这28份样本被用于搭载有约44000个探针的全基因组寡核苷酸微阵列进行杂交，该芯片可覆盖25585个带有基因符号的非冗余基因（附加文件1）。本研究同时使用同一份RNA样本开展定量反转录聚合酶链反应（RT-PCR）实验。