Unknown,Transcriptomics,Genomics,Proteomics

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Ectodysplasin regulates the lymphotoxin-beta pathway for hair differentiation

ABSTRACT: To find developmental stage specific genes controled by EDA Keywords: development or differentiation design,genetic modification design Triplicated for all samples

ORGANISM(S): Mus musculus

SUBMITTER: Yulan Piao

PROVIDER: E-GEOD-6952 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Ectodysplasin regulates the lymphotoxin-beta pathway for hair differentiation.

Cui Chang-Yi CY Hashimoto Tsuyoshi T Grivennikov Sergei I SI Piao Yulan Y Nedospasov Sergei A SA Schlessinger David D

Proceedings of the National Academy of Sciences of the United States of America 20060531 24

Mutations in the EDA gene cause anhidrotic/hypohidrotic ectodermal dysplasia, a disorder characterized by defective formation of hair, sweat glands, and teeth in humans and in a mouse model, "Tabby" (Ta). The gene encodes ectodysplasin, a TNF ligand family member that activates the NF-kappaB-signaling pathway, but downstream targets and the mechanism of skin appendage formation have been only partially analyzed. Comparative transcription profiling of embryonic skin during hair follicle developme ...[more]

PMID: 16738056

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Project description:The gold standard for examining pluripotency of stem cells is to see whether cells can contribute to entire body of animals. Here we show that the increased frequency of Zscan4 activation in mouse ES cells not only enhances, but also maintains their developmental potency in long-term cell culture. As the potency increases, even a whole animal can be produced from a single ES cell injected into 4N blastocyst at unusually high success rate. Although Zscan4-activated cells express genes that are also expressed in 2-cell stage mouse embryos, transiently Zscan4-activated state of ES cells is not associated with the high potency of ES cells. It is thus concluded that ES cells acquire higher potency by going through transient Zscan4 activation state more frequently than the regular state. Taken together, our results indicate that frequent activation of Zscan4 can rejuvenate pluripotent stem cells. MC1 ES cells (for pZscan4-Emerald transfection) and R26R6 ES cells (for pZscan4-Cre ERT2 transfection) were grown on gelatin in 6-well plate, 5x10^5cells in suspension were transfected with 1ug of linearized pZscan4-Emerald vector or pZscan4-Cre ERT2 vector using Effectene (QIAGEN) according manufacturer protocol, and plated in gelatin coated 100 mm dishes. Cells were selected with 5 ug/ml blasticidin, colonies were picked on the 8th day, expanded and frozen for further analysis. DNA microarray analysis of pZscan4-Emerald cells was carried out as described. In brief, universal Mouse Reference RNA (Stratagene) were labeled with Cy5-dye, mixed with Cy3-labeled samples, and used for hybridization on the NIA Mouse 44K Microarray v2.2 (manufactured by Agilent Technologies #014117). The intensity of each gene feature was extracted from scanned microarray images using Feature Extraction 9.5.1.1 software (Agilent Technologies). Microarray data analyses were carried out by using an application developed in-house to perform ANOVA and other analyses (NIA Array Analysis software; http://lgsun.grc.nia.nih.gov)

Project description: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 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)

Dataset Information

Ectodysplasin regulates the lymphotoxin-beta pathway for hair differentiation

Publications

Ectodysplasin regulates the lymphotoxin-beta pathway for hair differentiation.

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