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Prevedel, J.

Publications and source records attributed to Prevedel, J..

3 recordsLinked to original sources

Live longitudinal imaging of meningeal cerebrovascular injury and its sequelae in adult zebrafish

Nearly 1.4 million people in the United States sustain a traumatic brain injury (TBI) each year, with almost half of those hospitalized for TBI developing long-term disability. For many patients, prolonged bleeding and inflammation from damaged vessels in the meninges result in long-lasting sequelae. Although their injured blood vessels regrow, the site of injury is full of inflammatory immune cells that may influence vascular function. Adult zebrafish have a thin, translucent skull and a mammalian-like meninges that is easily imaged in living animals. We have established a novel adult zebrafish model to investigate vessel-immune cell interactions after meningeal cerebrovascular injury (mCVI). We use carefully calibrated sonication to rupture meningeal blood vessels without breaching the skull or causing damage to the underlying brain. By performing longitudinal live imaging of intubated adult fish we observe vascular regrowth and immune responses to mCVI over time in the same animal with unprecedented resolution allowing measurement of blood flow, dynamics of vessel regrowth, and interactions between individual immune and vascular cells. This newly developed zebrafish model provides a powerful tool for longitudinal live imaging of meningeal immune cell-vascular interactions after cerebrovascular injury, opening the door to new insights into chronic neuroinflammatory disease.

neuroscience↗

A novel transgenic reporter to study vertebrate epigenetics

Epigenetic reprogramming contributes to the generation of cellular diversity during vertebrate development but the mechanisms directing this are still not well understood. Large-scale genetic screens have been highly successful in identifying epigenetic regulatory genes in invertebrates such as worms and flies, but similar large-scale genetic screens to identify epigenetic regulators have not been carried out in vertebrates. Here we report a newly generated "EpiTag" zebrafish transgenic reporter line that permits easy cellular-level visualization of epigenetic silencing or activation in living animals during development, gametogenesis, and regeneration. We use the EpiTag reporter to carry out an F3 ENU mutagenesis screen for epigenetic silencing or activating mutants, identifying relevant vertebrate tissue-specific epigenetic regulatory genes including a new epigenetic model for metabolic dysfunction-associated fatty liver disease (MAFLD). The EpiTag reporter line represents a powerful new tool for genetic and experimental analysis of tissue-specific epigenetic gene regulation in vertebrates. One Sentence SummaryEpiTag transgenic zebrafish provide a powerful new tool for visualizing and studying epigenetic regulation in living vertebrate animals.

genomics↗

Comprehensive 3D imaging of whole zebrafish using a water-based clearing reagent for hard tissues

Zebrafish (Danio rerio) is a valuable model organism for studying developmental processes due to its external development and the optical clarity of its embryos and larvae. However, as development proceeds zebrafish form increasingly opaque tissues that impede visualization of deep tissues and structures. Although tissue clearing methods have been applied to facilitate imaging at these later stages, most of these methods have limited ability to clear dense tissues such as bone and cartilage, cause significant morphological distortion, and/or result in loss of fluorescent signal when used for imaging of fluorescent transgenes, dye stained animals, or specimens generated using immunofluorescence or fluorescence in situ hybridization methods. Here, we report a novel imaging technique using a recently developed clearing reagent called LUCID that makes it possible to capture the complete cellular-resolution 3D structures of larval and juvenile zebrafish. We show that LUCID clears dense tissues such as pharyngeal cartilage in juvenile animals and even tooth bone in adults without causing either significant morphological distortion or significant loss of signal from transgene-driven fluorescent proteins, fluorescent nuclear DNA or actin staining dyes, or whole mount in situ hybridization chain reaction (HCR) fluorescence. Using this new approach it is possible to perform complete high-resolution 3D imaging of whole fluorescently stained animals, even deep internal regions, providing a novel tool for elucidating the complex internal structures of developing zebrafish.

developmental biology↗