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Adv Exp Med Biol. 2013;789:15-20. doi: 10.1007/978-1-4614-7411-1_3.

Hypoxia-induced cerebral angiogenesis in mouse cortex with two-photon microscopy.

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Center for Frontier Science and Engineering, University of Electro-Communications, 1-5-1 Chofugaoka, Chofu, 182-8585, Tokyo, Japan.
Molecular Imaging Center, National Institute of Radiological Sciences, Chiba, Japan.
Molecular Imaging Center, National Institute of Radiological Sciences, Chiba, Japan.
Department of Neurology, Keio University School of Medicine, Tokyo, Japan.


To better understand cellular interactions of the cerebral angiogenesis induced by hypoxia, a spatiotemporal dynamics of cortical microvascular restructuring during an exposure to continuous hypoxia was characterized with in vivo two-photon microscopy in mouse cortex. The mice were prepared with a closed cranial window over the sensory-motor cortex and housed in 8-9 % oxygen room for 2-4 weeks. Before beginning the hypoxic exposure, two-photon imaging of cortical microvasculature was performed, and the follow-up imaging was conducted weekly in the identical locations. We observed that 1-2 weeks after the onset of hypoxic exposure, a sprouting of new vessels appeared from the existing capillaries. An average emergence rate of the new vessel was 15 vessels per unit volume (mm(3)). The highest emergence rate was found in the cortical depths of 100-200 μm, indicating no spatial uniformity among the cortical layers. Further, a leakage of fluorescent dye (sulforhodamine 101) injected into the bloodstream was not detected, suggesting that the blood-brain barrier (BBB) was maintained. Future studies are needed to elucidate the roles of perivascular cells (e.g., pericyte, microglia, and astroglia) in a process of this hypoxia-induced angiogenesis, such as sprouting, growth, and merger with the existing capillary networks, while maintaining the BBB.

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