Transcription profiling of mouse cornea from three strains reveal central corneal thickness is a genetic dependent trait among inbred strains of mice
ABSTRACT: Central corneal thickness (CCT) exhibits broad variability. We determined the corneal gene expression profile three mouse strains with distinct corneal thickness: C57BLKS/J (88.6 um), SJL/J (123.5 um), and C57BL/6J (100.1 um). Experiment Overall Design: Enucleated eyes from 4 month old mice were dissected in phosphate-buffered saline and a punch of central cornea was collected utilizing a 2-mm biopsy punch. Two central corneal punches (left and right eyes) were pooled from each mouse to form one sample; three samples were analyzed per strain.
Project description:Central corneal thickness (CCT) exhibits broad variability. We determined the corneal gene expression profile three mouse strains with distinct corneal thickness: C57BLKS/J (88.6 um), SJL/J (123.5 um), and C57BL/6J (100.1 um). Keywords: Strain comparison Overall design: Enucleated eyes from 4 month old mice were dissected in phosphate-buffered saline and a punch of central cornea was collected utilizing a 2-mm biopsy punch. Two central corneal punches (left and right eyes) were pooled from each mouse to form one sample; three samples were analyzed per strain.
Project description:Transplantation of ex vivo expanded limbal stem cells (LSC) is the main treatment for limbal stem cell deficiency though the clinical problem of donor tissues shortage. Recently, as the development of tissue engineering, embryonic stem cells (ESC) derived corneal epithelial-like cells (ESC-CEC) has become a new direction to this issue.Our group successfully induced ESC into corneal epithelial-like cells, and in the present study we explored various aspects of physiological properties of ESC-CEC. The experiment included three samples: hES, the human embryonic stem cell line H1, RA_SB, the corneal epithelial-like cells derived from hES by differentiation with RA and SB, epithelial_cell, the primary human limbal stem cells from cadaver eyes. hES, the human embryonic stem cell line H1, RA_SB, the corneal epithelial-like cells derived from hES by differentiation with RA and SB, epithelial_cell, the primary human limbal stem cells from cadaver eyes.
Project description:To identifycandidate PITX2-dependent genes during corneal development, microarray analysis was used to profile gene expression in corneas taken from flash-frozen e12.5 mouse embryos of control (Pitx2+/-) and temporal knockout Pitx2-deficient embryos. Overall design: To identify candidate PITX2-dependent targets, gene was independently measured in corneal tissue isolated from four embryos of each geneotype. Corneal tissue isolated from both eyes of a given embryo were pooled and constituted an individual sample pool.
Project description:PPARgamma null (PpargΔ/Δ) mice present a generalized lipoatrophy and a dramatic skin phenotype, characterized by delayed hair morphogensis and the appearance, at adult age, of severe inflammatory infiltration. To investigate the molecular mechanisms underlying the delayed hair morphogenesis observed in PpargΔ/Δ mice, we used microarrays to detail the global program of gene expression in full thickness skin of PpargΔ/Δ mice and respective control mice at embryionic stage E17.5. Overall design: We examined 2 experimental groups: PpargΔ/Δ and control mice at E17.5. For each experimental group we analysed the transcriptional profile of 4 individual RNA from full thickness skin.
Project description:Differences in regional protein expression within the human retina may explain molecular predisposition of specific regions to ophthalmic diseases like age-related macular degeneration, cystoid macular edema, retinitis pigmentosa, and diabetic retinopathy. To quantify protein levels in the human retina and identify patterns of differentially-expressed proteins, we collected foveal, macular, and peripheral retina punch biopsies from healthy donor eyes and analyzed protein content by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Project description:Temporal coordination of developmental programs is necessary for normal ontogeny, but the mechanism by which this is accomplished is poorly understood. We have previously shown that two components of the Mediator CDK8 module, CENTER CITY (CCT/MED12) and GRAND CENTRAL (GCT/MED13), are required for timing of pattern formation during embryogenesis in Arabidopsis. Here, we performed global gene expression analyses of wild-type, cct-1, and gct-2 seedlings (above-ground portions only) to help analyze their post-embryonic phenotypes. Our results suggest that MED12 and MED13 act as global regulators of developmental timing by fine-tuning expression of temporal regulatory genes. Seeds of wild-type Col-0, cct-1, and gct-2 were sown in Fafard #2 soil. Seedlings (above-ground portions only) were harvested when the first two leaf primordia were 1 mm in length, which was at day 7 for wild-type seedlings and day 9 for the two mutants. 75-100 seedlings were used for each of three biological replicates.
Project description:While considerable progress has been made towards understanding the complex processes and pathways that regulate human wound healing, regenerative medicine has been unable to develop therapies that coax the natural wound environment to heal scar-free. The inability to induce perfect skin regeneration stems partly from our limited understanding of how scar-free healing occurs in a natural setting. Here we have investigated the wound repair process in adult axolotls and demonstrate that they are capable of perfectly repairing full thickness excisional wounds made on the flank. In the context of mammalian wound repair, our findings reveal a substantial reduction in hemostasis, reduced neutrophil infiltration and a relatively long delay in production of new extracellular matrix (ECM) during scar-free healing. Additionally, we test the hypothesis that metamorphosis leads to scarring and instead show that terrestrial axolotls also heal scar-free, albeit at a slower rate. Analysis of newly forming dermal ECM suggests that low levels of fibronectin and high levels of tenascin-C promote regeneration in lieu of scarring. Lastly, a genetic analysis during wound healing comparing epidermis between aquatic and terrestrial axolotls suggests that matrix metalloproteinases may regulate the fibrotic response. Our findings outline a blueprint to understand the cellular and molecular mechanisms coordinating scar-free healing that will be useful towards elucidating new regenerative therapies targeting fibrosis and wound repair. We used microarray analysis to determine the gene expression changes that take place during scar free wound healing in aquatic and terrestrial axolotl salamanders. Epidermal tissue was harvested using a 4mm biopsy punch. Two wounds were made along the flank and posterior to the forelimbs. Harvested epidermis was pooled for each animal. Four biological replicates were collected from uninjured epidermis (D0) and at 1, 3, and 7 days post injury.
Project description:Examine the effect of stimulation stress produced by chemical burn to the mice cornea in the lacrimal gland over a time course (0.5 - 360 hours) in comparison with the non-stimulated control animals. Using silver nitrate applicator (silver nitrate 37.5 mg and potassium nitrate 12.5 mg, Graham-Field,Inc.,) do two burns per eye, each with a separate applicator. For animals observed for more than 24 hours, eyes were then treated with gentamicin ointment to minimize infection. Total RNA were extracted from both burn and control eyes and were covalently coupled separately with Cy5 and Cy3 monoreactive fluors. The Cy5 and Cy3 labelled RNA were hybridized to the microarray. Fluorescent array images were collected for both Cy3 and Cy5 with ScanArray 4000XL and image intensity data were extracted and analyzed with Imagene analysis software. Ref:Fang Y, Choi D, Searles RP, Mathers WD.A time course microarray study of gene expression in the mouse lacrimal gland after acute corneal trauma.Invest Ophthalmol Vis Sci. 2005 Feb;46(2):461-9. Keywords = Corneal injury Keywords = mouse Keywords = lacrimal gland Keywords = gene microarrays Keywords: time-course