Intravital imaging of mouse embryos
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2020-04-10
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<jats:p>Embryonic development is a complex process that is unamenable to direct observation. In this study, we implanted a window to the mouse uterus to visualize the developing embryo from embryonic day 9.5 to birth. This removable intravital window allowed manipulation and high-resolution imaging. In live mouse embryos, we observed transient neurotransmission and early vascularization of neural crest cell (NCC)–derived perivascular cells in the brain, autophagy in the retina, viral gene delivery, and chemical diffusion through the placenta. We combined the imaging window with in utero electroporation to label and track cell division and movement within embryos and observed that clusters of mouse NCC-derived cells expanded in interspecies chimeras, whereas adjacent human donor NCC-derived cells shrank. This technique can be combined with various tissue manipulation and microscopy methods to study the processes of development at unprecedented spatiotemporal resolution.</jats:p>
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Huang, Qiang, Malkiel A Cohen, Fernando C Alsina, Garth Devlin, Aliesha Garrett, Jennifer McKey, Patrick Havlik, Nikolai Rakhilin, et al. (2020). Intravital imaging of mouse embryos. Science, 368(6487). pp. 181–186. 10.1126/science.aba0210 Retrieved from https://hdl.handle.net/10161/20403.
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Scholars@Duke
Yubin Kang
Blanche Capel
In mammals, the primary step in male sex determination is the initiation of testis development in the bipotential gonad primordium. This step depends on the Y-linked male sex-determining gene, Sry. Expression of Sry in the XY gonad, or as a transgene in an XX gonad, leads to the differentiation of Sertoli cells. Failures in Sertoli cell differentiation in the XY gonad result in sex reversal and ovary formation. We are also interested in the biology of germ cells -- the cells that give rise to eggs and sperm. I have had a longstanding interest in the communication between Sertoli cells and germ cells in fetal life and afterwards, once the seminiferous epithelium is established. In adult life, each Sertoli cell communicates with germ cells at multiple stages of development from spermatogonial stem cells located at their base to elongated spermatids, released at their apical surface. How can Sertoli cells direct specific information to each of the germ cells wedged between their cell membranes? This problem previously seemed unapproachable, because it was so difficult to figure out where to anchor the analysis, and the cost of transgenics seemed prohibitive. However, recently we have been collaborating with a colleague in my department (Scott Soderling) who has designed an AAV system to deliver CRISPR and tag genetic loci in vivo with BioID for proximity protein labeling. We found that we can deliver the backbone AAV to Sertoli cell within seminiferous tubules with high efficiency by injection into the rete testis. Combining expertise of our labs provides an opportunity to do a similar analysis in Sertoli cells with the goal of identifying localized communication between Sertoli cells and the germ cells they support.
Debra Lynn Silver
How is the brain assembled and sculpted during embryonic development? Addressing this question has enormous implications for understanding neurodevelopmental disorders affecting brain size and function. In evolutionary terms, our newest brain structure is the cerebral cortex, which drives higher cognitive capacities. The overall mission of my research lab is to elucidate genetic and cellular mechanisms controlling cortical development and contributing to neurodevelopmental pathologies and brain evolution. We study neural progenitors, essential cells which generate neurons and are the root of brain development. We are guided by the premise that the same mechanisms at play during normal development were co-opted during evolution and when dysregulated, can cause neurodevelopmental disease.
My research program employs a multifaceted strategy to bridge developmental neurobiology, RNA biology, and evolution. 1) We investigate how cell fates are specified, by studying how progenitor divisions influence development and disease. 2) We study diverse layers of post-transcriptional regulation in neural progenitors. We investigate RNA binding proteins implicated in development and neurological disease. Using live imaging, we also investigate how sub-cellular control of mRNA localization and translation influences neural progenitors. 3) A parallel research focus is to understand how human-specific genetic changes influence species-specific brain development. Our goal is to integrate our efforts across these three major lines of research to understand the intricacies controlling brain development.
Xiling Shen
Dr. Shen’s research interests lie at precision medicine and systems biology. His lab integrates engineering, computational and biological techniques to study cancer, stem cells, microbiota and the nervous system in the gut. This multidisciplinary work has been instrumental in initiating several translational clinical trials in precision therapy. He is the director of the Woo Center for Big Data and Precision Health (DAP) and a core member of the Center for Genomics and Computational Biology (GCB).
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