The long-term objective of this project is to better understand the mechanisms regulating brain regionalization during early development, using zebrafish embryos as a model system. Large-scale mutagenesis screens in zebrafish have revealed a network of genes, which play a role in patterning of the brain. However, the dynamics of this network is only partly understood. New insights may be obtained by combining ideas and approaches from developmental genetics and neurophysiology. The central hypothesis of this project is that calcium signals play a key role in the signaling pathways that pattern the brain. Calcium signals can spread rapidly over large distances within the embryo and are capable of regulating a variety of cellular events including post-translational modification of signaling proteins, secretion, contraction, and gene expression. Moreover, various lines of evidence suggest a role of calcium in the midline signaling pathway, which induces the neural tube floor plate and ventral brain regions. To further test the role of calcium in midline signaling, calcium patterns will be manipulated and effects on neural tube and brain development will be characterized. In addition, the interaction between calcium and components of the midline signaling pathway will be examined. Obtained results will give a first view of calcium interacting with signaling pathways in the developing forebrain. A broader impact of this project is the establishment of full-time research within the Bioimaging Facility at Brown University. This multi-user facility provides training in microscopy to undergraduate students, graduate students, post-docs, staff, faculty, and visiting scientists. Local high schools regularly tour the facility, and for many high school students this is their first visit to a university, and their first contact with scientific research. In addition, the facility provides free consultation on microscopy and image analysis to the local community including other universities, small businesses, and nearby hospitals. A full-time research program at the Bioimaging Facility will affect the outreach potential of this facility, in part by enhancing the infrastructure of this facility, and in part by bringing in investigators who will teach microscopy and image analysis.
