Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:Analysis of gene expression differences in three types of mouse motor neurons: (1) those harvested from E13.5 mouse embryos; (2) those derived from embryonic stem cells by directed differentiation; and (3) those made from mouse embryonic fibroblasts by transcription factor reprogramming. Hb9::GFP+ mouse motor neurons were obtained by three methods [(1) dissection of Hb9::GFP-transgenic E13.5 mouse embryo spinal cord; (2) directed differentiation of an Hb9::GFP-transgenic mouse embryonic stem cell line, V6.5; (3) reprogramming of Hb9::GFP-transgenic mouse embryonic fibroblasts by overexpression of 10 transcription factors]. Each type of motor neuron was purified by FACS and harvested in Trizol. Total RNA was purified and prepared for hybridization onto Illumina MouseRef-8.
Project description:Analysis of gene expression differences in three types of mouse motor neurons: (1) those harvested from E13.5 mouse embryos; (2) those derived from embryonic stem cells by directed differentiation; and (3) those made from mouse embryonic fibroblasts by transcription factor reprogramming.
Project description:We use comprehensive and unsupervised transcriptome analyses to provide molecular classifications of sensory neurons in the mouse geniculate ganglion. 96 neurons were isolated on a C1 Fluodigm chip, underwent RNA-Seq, and iteratively clustered into sub-classes.