Abstract
Live confocal microscopy of vital fluorescent markers, expressed in mouse embryonic tissues, is a powerful and exciting method to study mammalian embryonic development. This chapter discusses imaging approaches to visualize and characterize dynamic changes of the yolk-sac vasculature and blood flow in mouse embryos. We describe static embryo-culture protocols, which allow maintaining early mouse embryos on the imaging stage for over 24 h. We also describe vital fluorescent-reporter lineages, which can be used to image the developing vasculature and characterize hemodynamics by tracking individual blood cells. Imaging approaches described in this chapter can be used to analyze cardiovascular defects in mutant animals and can provide insights into understanding how genetic signaling pathways and physiological inputs regulate development.
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References
Thoma, R. (1893) Untersuchungen über die Histogenese und Histomechanik des Gefässsystems. Stuttgart: Ferdinand Enke.
Lucitti, J.L., Jones, E.A.V., Huang, C., Chen, J., Fraser, S.E., Dickinson, M.E. (2007) Vascular remodeling of the mouse yolk sac requires hemodynamic force. Development 134, 3317–3326.
Hadjantonakis, A.K., Dickinson, M.E., Fraser, S.E., Papaioannou, V.E. (2003) Technicolour transgenics: imaging tools for functional genomics in the mouse. Nat Rev Genet 4, 613–625.
Lichtman, J.W., Fraser, S.E. (2001) The neuronal naturalist: watching neurons in their native habitat. Nat Neurosci 4 Suppl, 1215–1220.
Kulesa, P.M. (2004) Developmental imaging: Insights into the avian embryo. Birth Defects Res C Embryo Today 72, 260–266.
Jones, E.A.V., Crotty, D., Kulesa, P.M., et al. (2002) Dynamic in vivo imaging of postimplantation mammalian embryos using whole embryo culture. Genesis 34, 228–235.
Jones, E.A.V., Baron, M.H., Fraser, S.E., Dickinson, M.E. (2004) Measuring hemodynamic changes during mammalian development. Am J Physiol Heart Circ Physiol 287, H1561–1569.
Larina, I., Shen, W., Kelly, O., Hadjantonakis, A., Baron, M., Dickinson, M. (2009) A membrane associated mCherry fluorescent reporter line for studying vascular remodeling and cardiac function during murine embryonic development. Anatomical Record 292, 333–341.
Larina, I.V., Sudheendran, N., Ghosn, M., et al. (2008) Live imaging of blood flow in mammalian embryos using Doppler swept source optical coherence tomography. Journal of Biomedical Optics 13, 0605061–3.
Larina, I.V., Ivers, S., Syed, S., Dickinson, M.E., Larin, K.V. (2009) Hemodynamic measurements from individual blood cells in early mammalian embryos with Doppler swept source OCT. Optics Letters 34, 986–988.
Fraser, S.T., Hadjantonakis, A-K., Sahr, K.E., et al. (2005) Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice. Genesis 42, 162–171.
Dyer, M.A., Farrington, S.M., Mohn, D., Munday, J.R., Baron, M.H. (2001) Indian hedgehog activates hematopoiesis and vasculogenesis and can respecify prospective neurectodermal cell fate in the mouse embryo. Development 128, 1717–1730.
Dirnagl, U., Villringer, A., Einhaupl, K.M. (1992) In vivo Confocal Scanning Laser Microscopy Of The Cerebral Microcirculation. J Microsc-Oxf 165, 147–157.
Kleinfeld, D., Mitra, P.P., Helmchen, F., Denk, W. (1998) Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex. Proc Natl Acad Sci USA 95, 15741–15746.
Acknowledgments
The study is supported by the National Institutes of Health (R01HL095586) and the American Heart Association (10SDG3830006).
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Larina, I.V., Dickinson, M.E. (2012). In Vivo Imaging of the Developing Mouse Embryonic Vasculature. In: Hoffman, R. (eds) In Vivo Cellular Imaging Using Fluorescent Proteins. Methods in Molecular Biology, vol 872. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-797-2_14
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DOI: https://doi.org/10.1007/978-1-61779-797-2_14
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