Project description:The foggy mutant zebrafish show severe deficits in dopaminergic neurons, thus is an important tool to understanding the development of these neurons, which undergo degeneration in Parkinson's disease. Molecular study reveals that the foggy gene encodes a regulator of transcription elongation. Biochemical study suggests that the repressive activity of the Foggy protein is lost in the foggy mutant. The genes whose expression is normally repressed by Foggy are likely to play important roles in dopamine neuron development. However, their identity is unknown. Finding out these target genes of Foggy will provide important insights into dopamine neuron development and regeneration. They will also serve as an entry point to elucidate the role of transcription elongation in controlling neuronal development. We will identify genes whose expression is altered in the foggy mutant. Their expression pattern and functional significance will be examined in zebrafish through in situ hybridization and morpholino antisense knockdowm methods. We hypothesize that the genes whose transcription is regulated by Foggy play a critical role in the development of dopaminergic neurons. The species noted above is Drosophila however we are using ZEBRAFISH. We will examine three stages: 24 hour post fertilization (hpf), 36 hpf, and 48 hpf. During these stages, the dopamine neuron deficits manifest in the foggy mutant. To identify genes whose expression is altered and important for dopamine neuron development, we shall dissect embryonic brain tissues from wildtype and the foggy mutant embryos at the stages mentioned above, and prepare RNA from them. Since the foggy gene is expressed ubiquitously and also required for proper retinal and cardiovascular development, we would like to compare the gene expression between the whole embryos of wildtype and foggy mutant, to identify all possible downstream targets. We are currently applying for a supplement to support the entire microarray experiment. As a pilot experiment, we would like to perform 6 array analysis as soon as possible. Thus, we will examine the 48 hours post fertilzation time point in this pilot experiment and it will be performed in triplicate. This dataset is composed of 36 hours post fertilization wild type and mutants and 12-somite stage Foggy knockdown.

Project description:The foggy mutant zebrafish show severe deficits in dopaminergic neurons, thus is an important tool to understanding the development of these neurons, which undergo degeneration in Parkinson's disease. Molecular study reveals that the foggy gene encodes a regulator of transcription elongation. Biochemical study suggests that the repressive activity of the Foggy protein is lost in the foggy mutant. The genes whose expression is normally repressed by Foggy are likely to play important roles in dopamine neuron development. However, their identity is unknown. Finding out these target genes of Foggy will provide important insights into dopamine neuron development and regeneration. They will also serve as an entry point to elucidate the role of transcription elongation in controlling neuronal development. We will identify genes whose expression is altered in the foggy mutant. Their expression pattern and functional significance will be examined in zebrafish through in situ hybridization and morpholino antisense knockdowm methods. We hypothesize that the genes whose transcription is regulated by Foggy play a critical role in the development of dopaminergic neurons. The species noted above is Drosophila however we are using ZEBRAFISH. We will examine three stages: 24 hour post fertilization (hpf), 36 hpf, and 48 hpf. During these stages, the dopamine neuron deficits manifest in the foggy mutant. To identify genes whose expression is altered and important for dopamine neuron development, we shall dissect embryonic brain tissues from wildtype and the foggy mutant embryos at the stages mentioned above, and prepare RNA from them. Since the foggy gene is expressed ubiquitously and also required for proper retinal and cardiovascular development, we would like to compare the gene expression between the whole embryos of wildtype and foggy mutant, to identify all possible downstream targets. We are currently applying for a supplement to support the entire microarray experiment. As a pilot experiment, we would like to perform 6 array analysis as soon as possible. Thus, we will examine the 48 hours post fertilzation time point in this pilot experiment and it will be performed in triplicate.

Dataset Information

RNA-Seq of mouse E12.5 brain regions

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