Project description:Medaka fish is a long standing genetic model organism from the 1930s. Uniquely amongstvertebrates Medaka fish can be routinely inbred from the wild (laboratory mice are inbred, butthis does not happen as a routine process from wild individuals), leading to a large number offully inbred wild-derived strains. As part of a broader collaboration I am part of a project toinbred over 100 wild derived strains of Medaka and use them in a similar manner toArabidopsis and Drosophila wild derived lines.To help explore the phenotyping possibilities in this context, we have taken alreadyestablished wild Medaka lines and done reciprocal F1 crosses between 3 strains, and have 4tissues, and a number of parental tissues, giving a total of 40 samples. By doing RNA-seq onthese tissues we do a number of things:(a) assess the level of allele specific expression in Medaka(b) test whether there is any imprinting (parent of origin) in fish. This is thought to not be thecase, but in fact has not been well tested (not least because getting truly inbred fish is hard)(c) assess the level of random allelic activation in brain(d) assess the feasibility for RNAseq based phenotyping in Medaka, providing preliminarydata for future proposals.This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:During development, the inherited DNA methylation patterns from the parental gametes needs to be remodeled into a state compatible with embryonic pluripotency. In Zebrafish, this remodeling is achieved by the maternal methylome becoming hypomethylated to match the paternal methylome. However, how this is achieved in medaka (another teleost fish) is currently not known. Moreover, how DNA methylation remodeling is impacted in hybrid organisms, and the effects this may have on their development, is also not known. Here we address these questions by generation whole genome bisulfite sequencing data for zebrafish, medaka and zebrafish medaka embryos.
Project description:We employed a transgenic strain of a small aquarium fish medaka (Oryzias latipes) that overexpresses a malignant melanoma driver gene. In this model, melanoma develops with 100 % penetrance. Using the medaka malignant melanoma model, we tested whether cisplatin and trametinib are able to suppress the transcriptomic changes induced by the transgene.
Project description:Using medaka fish embryo model, toxic effects of silver nanocolloids (SNC, 3.8M-BM-11.0nm diameter) on developmental morphology and gene expression profile were investigated. SNC caused morphological changes in embryos including cardiovascular malformations, ischemia, underdeveloped central nervous system and eyes, and kyphosis at exposures of 0.5mg/L and 1.0mg/L.. To determine in vivo distribution of SNC, medaka embryos were exposed to 0.5mg/L for 6 days and subjected to ICP-OES analyses. Silver was detected in medaka embryos and chorion at levels of 16.6M-BM-19.3pg and 720M-BM-129pg, respectively. TEM analyses showed SNC adhered to the chorion surface and inside the chorion. On investigation of oxidative mechanism, NAC (0.05mM) rescued all embryos by 96-hr post treatment, while 0.5mM GSH did not. NAC blocked lipid peroxidation. Furthermore, medaka oligo DNA microarray and qRT-PCR were used for gene expression profiling in embryos exposed for 48-hr to 0.05mg/L SNC. Six genes relative to embryogenesis and morphogenesis such as ctsL, Tpm1, RBP, mt, atp2a1 and hox6b6 turned out to be affected and their involvement to the above malformations was implied. In conclusion, SNC causes gross malformations in the cardiovascular and central nervous systems in developing medaka embryos through potentially SNC-affected differential expression of a series of genes related to oxidative stress, embryonic cellular proliferation, and morphological development. Three of the stage-21 medaka embryos per sample were exposed in triplicate to 0mg/L (control) and 0.05mg/L SNC for 48 hours. Therefore, for each exposure condition nine of the stage-21 embryos were prepared and used in the SNC exposure test.
Project description:Medaka fish were exposed to several concentrations of humic acid. Changes in liver protein profiles of control and exposed fish were studied using LC-MS/MS analysis.
Project description:The “Fish Acute Toxicity Test” in the “OECD Guideline for Testing of Chemicals” is an essential test for environmental toxicity. Here we have tried to evaluate the physiology of medaka during this test procedure by using genomics. Genomics technology can provide genome-wide expression profiles of mRNA, and these profiles correspond to the physiology of organisms. Thus, a comparison of mRNA expression profiles gives information on the reproducibility of experimental conditions. Expression profiles of mRNA were measured for medakas maintained within the allowable range of the test conditions and also under extreme conditions beyond the guidelines limits. We confirmed the high physiological reproducibility of medaka kept in the recommended conditions of the “Fish Acute Toxicity Test” in the “OECD Guideline for Testing of Chemicals" from the expression profiles obtained under all experimental conditions except for the type of feeding.
Project description:Medaka fish were exposed to several concentrations of humic acid. Changes in plasma protein profiles of control and exposed fish were studied using LC-MS/MS analysis.
Project description:Using medaka fish embryo model, toxic effects of silver nanocolloids (SNC, 3.8±1.0nm diameter) on developmental morphology and gene expression profile were investigated. SNC caused morphological changes in embryos including cardiovascular malformations, ischemia, underdeveloped central nervous system and eyes, and kyphosis at exposures of 0.5mg/L and 1.0mg/L.. To determine in vivo distribution of SNC, medaka embryos were exposed to 0.5mg/L for 6 days and subjected to ICP-OES analyses. Silver was detected in medaka embryos and chorion at levels of 16.6±9.3pg and 720±29pg, respectively. TEM analyses showed SNC adhered to the chorion surface and inside the chorion. On investigation of oxidative mechanism, NAC (0.05mM) rescued all embryos by 96-hr post treatment, while 0.5mM GSH did not. NAC blocked lipid peroxidation. Furthermore, medaka oligo DNA microarray and qRT-PCR were used for gene expression profiling in embryos exposed for 48-hr to 0.05mg/L SNC. Six genes relative to embryogenesis and morphogenesis such as ctsL, Tpm1, RBP, mt, atp2a1 and hox6b6 turned out to be affected and their involvement to the above malformations was implied. In conclusion, SNC causes gross malformations in the cardiovascular and central nervous systems in developing medaka embryos through potentially SNC-affected differential expression of a series of genes related to oxidative stress, embryonic cellular proliferation, and morphological development.
Project description:Alkalinity stress is considered to be one of the major stressors for fish in saline-alkali water. Thus, it is of great significance from both aquaculture and physiological viewpoint to understand the molecular genetic response of aquatic organisms to alkalinity stress. The objective of this study is to determine genome-wide gene expression profiles to better understand the physiology response of medaka (Oryzias latipes) to high carbonate alkalinity stress. In lab-based cultures, adult fish were exposed to freshwater and high carbonate alkalinity water .We designed a microarray containing 26429 oligonucleotides and describe our experimental results for measuring gene expression changes in the gill of carbonate alkalinity stress exposed fish. The fish were exposed to freshwater (FW) and high carbonate alkalinity water (AW) for 96h, each with three replicates.
Project description:To examine the seasonal adaptation, we compared the gene expression of eyes between SL (short-day and low-temperature conditions: 10 h light/14 h dark and 8 °C) and LD (long-day and warm-temperature conditions: 14 h light/10 h dark and 26 °C) conditions in Medaka fish (Oryzias latipes).