Hierarchical imaging and computational analysis of three-dimensional vascular network architecture in the entire postnatal and adult mouse brain

Thomas Wälchli, Jeroen Bisschop, Arttu Miettinen, Alexandra Ulmann-Schuler, Christoph Hintermüller, Eric P. Meyer, Thomas Krucker, Regula Wälchli, Philippe P. Monnier, Peter Carmeliet, Johannes Vogel, Marco Stampanoni

Published: 2021-09-02 DOI: 10.1038/s41596-021-00587-1

vascular network

mouse brain

3D imaging

computational analysis

developmental stage

AI 解读

Extended

Extended Data Fig. 1 Intracardial resin perfusion and brain dissection for adult (P60) animals.

The main steps of resin perfusion and brain dissection are shown as photographs for the perfusion and brain dissection of adult (P60) mice. a - c Fixation ( a) , surgical opening ( b , c ) and site of needle insertion ( c ) for intracardial perfusion (via the left ventricle) with 10–20 ml ACSF containing 25,000 U/L Heparin, followed by 4% PFA in PBS, and then by the polymer resin PU4ii (vasQtec, Zürich, Switzerland), all infused at the same rate (8 ml/min and 100-120 mmHg) in an adult (P60) mouse (see also Supplementary Video 1 ). d Successful perfusion is indicated by a bluish aspect of the animal, which can be easiest observed at the paws (white arrow) or the snout (white arrowhead). e , f Resin cast of a P60 animal after soft tissue maceration, and before ( e ) and after ( f ) decalcification of bone structures. f - h The cerebral vasculature is sharply dissected from the extracranial vessels resulting in the isolated brain vascular corrosion cast of the P60 mouse brain ( g , h ). Scale bars, 10 mm ( e - h ).

Extended Data Fig. 2 Validation of vascular corrosion casting in postnatal (P10) and adult (P60) mice using light microscopy (LM) and SEM.

a - f Quality control of the vascular corrosion casts of P10 and P60 WT mice by visual inspection by LM ( a ), and by SEM ( b - f ). a Light microscopy image of the entire brain vasculature of the P60 WT mice (gold sputtered for SEM). b SEM image of the entire brain vascular corrosion cast illustrating the dense vascular network including blood vessels of all sizes with recognizable vascular anatomy. c - f The entire brain was uniformly filled with resin and the lumen of the vessels imprinted in the casting material with the finest details. e The capillary network in the cortex is well defined and devoid of interruptions. f Note the elongated cellular and nuclear imprints on the large vessel that are also typical for good-quality vascular corrosion casts in adult mice 105 . Scale bars, 4 mm ( a ); 4 mm ( b ); 50 μm ( c ); 50 μm ( d ), 100 μm ( e ), 100 μm ( f ). a,d - f reproduced with permission from ref. 39. b reproduced with permission from ref. 100 .

Extended Data Fig. 3 Global vascular network morphometry: increased vascular volume fraction in various regions of the adult (P60) Nogo-A -/- versus postnatal (P10) Nogo-A -/- mouse brain.

Genetic deletion of Nogo-A leads to increased vascular volume fraction in various regions of the postnatal mouse brain. a - c Computational 3D reconstructions of μCT scans of vascular networks of P10 and P60 Nogo-A -/- mice displayed with color-coded vessel thickness. The increased vessel density in the Nogo-A -/- animals, as compared to the WT animals (Fig. 5 ) in the cortices ( a ), hippocampi ( b ) and superior colliculi ( c ) is obvious. Color bar indicates vessel radius. The boxed areas are enlarged at right. (n = 11 for P10 Nogo-A -/- ; n = 13 for P60 Nogo-A -/- animals; and in average three ROIs per animal and brain region were used). Scale bars: 100 µm ( a - c overviews); 50 µm ( a - c zooms).

Extended Data Fig. 4 Local vascular network topology: increased vascular volume fraction of the adult (P60) Nogo-A -/- versus postnatal (P10) Nogo-A -/- mouse brain mainly found at the capillary level.

Genetic deletion of Nogo-A leads to increased vascular volume fraction in various regions of the P10 mouse brain. a Computational 3D reconstruction of μCT images of vascular networks of P10 and P60 Nogo-A -/- mice separated for noncapillaries (magenta) and capillaries (green). The increased density of capillaries (green) in the P60 and P10 Nogo-A -/- samples is evident when compared to the WT animals (Fig. 7 ), noncapillaries (magenta) appear largely unchanged. The boxed areas are enlarged at right ( a - c ). Scale bars: 100 mm ( a - c , overviews); 50 mm ( a - c , zooms).

Extended Data Fig. 5 Local vascular network topology: visualization and quantification of vascular branch point diameter and vascular branch point density in the hippocampus of the adult (P60) WT versus the postnatal (P10) WT mouse brain.

The vascular branch point diameter and vascular branch point density are significantly increased in the P60 as compared to the P10 mouse brain hippocampus. a Scheme depicting the definition of vascular branch points (see Fig. 8b for details) b Histogram showing the distribution of branch point diameter plotted against the branch point density in P10 WT and P60 WT animals. P60 WT animals show an increased branch point density as compared to P10 WT (bin width = 0.38 μm; number of bins = 40). Black dashed line marks the separation between capillaries (< 7 μm) and non-capillaries (≥7 μm), as defined in Fig. 6 . c,d Computational 3D reconstructions of μCT images of vascular networks in the hippocampi of P10 WT and P60 WT mice with visualizations of the vessel branch points displayed as dots, separated for noncapillaries (magenta) and capillaries (green). The higher density of branch points in the P60 WT hippocampi especially at the capillary level (green) is obvious, a slight increase of branch point density can be observed at the noncapillary level (magenta). The boxed areas are enlarged at right. e – g Quantitative analysis of the branch point density for all vessels ( e ), noncapillaries ( f ), and capillaries ( g ) in P10 WT and P60 WT hippocampi by local morphometry analysis. The significant increase of the branch point density for all vessels in the P60 WT animals ( e ) was mainly due to a significant increase at the level of capillaries ( g ) and in part due to a significant increase at the level of noncapillaries ( f ). In average, n = 2-7 animals were used for the hippocampi and in average three ROIs per animal and brain region were used. All data are shown as mean distributions where the open dot represents the mean. Boxplots indicate the 25% to 75% quartiles of the data. The shaded blue and red areas indicate the SD. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 100 µm ( c - d overviews); 50 µm ( c - d zooms).

Extended Data Fig. 6 Local vascular network topology: visualization and quantification of vascular branch point diameter and vascular branch point density in the superior colliculus of the adult (P60) WT versus the postnatal (P10) WT mouse brain.

The vascular branch point diameter and vascular branch point density are significantly increased in the P60 as compared to the P10 mouse brain superior colliculus. a Scheme depicting the definition of degree 3 and 4 vascular branch point (see Fig. 8b for details). b Histogram showing the distribution of branch point diameter plotted against the branch point density in P10 WT and P60 WT animals. P60 WT animals show an increased branch point density as compared to P10 WT mice mainly at the capillary level (bin width = 0.38; number of bins = 40). Black dashed line marks the separation between capillaries (< 7 μm) and noncapillaries (≥7 μm), as defined in Fig. 6 . c , d Computational 3D reconstructions of μCT images of vascular networks of the superior colliculi of P10 WT and P60 WT mice with visualizations of the vessel branch points displayed as dots, separately for noncapillaries (magenta) and capillaries (green). The higher density of branch points in the P60 WT superior colliculi especially at the capillary level (green) is obvious, a slight increase of branch point density can be observed at the noncapillary level (magenta). The boxed areas are enlarged at right. e – g Quantitative analysis of the branch point density for all vessels ( e ), noncapillaries ( f ), and capillaries ( g ) in P10 WT and P60 WT superior colliculi by local morphometry analysis. The significant increase of the branch point density for all vessels in the P60 WT animals ( e ) was mainly due to a significant increase at the level of capillaries ( g ) and in part due to a significant increase at the level of noncapillaries ( f ). In average, n = 2-4 animals were used for the superior colliculus; and in average three ROIs per animal and brain region were used. All data are shown as mean distributions where the open dot represents the mean. Boxplots indicate the 25% to 75% quartiles of the data. The shaded blue and red areas indicate the SD. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 100 μm ( c , d , overviews); 50 μm ( c , d , zooms).

Extended Data Fig. 7 Resin PU4ii vascular corrosion casting combined with immunofluorescence.

Visualization by fluorescence light microscopy of the brain vasculature after the addition of fluorescent pigments to the PU4ii resin solution in the P60 mouse brains with preserved/non-macerated soft tissue ( a - f ), co-labeled with red fluorescent myelin stainings ( g - j ). Overviews ( a,b , d,e ) and coronal cortical sections ( c , f ) of P60 mouse brains with yellow/green ( a , b , c ) and UV-blue ( d , e,f ) fluorescent dyes (vasQtec) added to the PU4ii resin showing the brain vasculature; overviews: dorsal overviews ( a , d ), ventral overviews ( b , e ), coronal sections ( c , f ). The selection of the brain regions analyzed here has been done on coronal sections in the structures selected using the Allen Mouse Brain Atlas. c , f , g - j Fluorescence light microscopy overviews of multiple brain regions in 150 μm coronal sections of a P60 mouse brain perfused with either yellow/green- ( c , h - j ) or UV-blue ( f , g ) fluorescent pigments added to the PU4ii resin solution, showing the cortical layers and part of the hippocampus ( c , f ). Fluorescence light microscopy of coronal sections in which the preserved/non-macerated soft tissue was labeled with a red fluorescent myelin stain (FluoroMyelin Red, Molecular Probes Inc) to visualize the corpus callosum and other highly myelinated brain structures (red). Scale bars, 1 mm ( a , b , d , e ); 500 μm ( c , d , g - j ). Excitation/emission of the yellow/green pigments at 450 nm/550 nm, excitation/emission of the UV-blue fluorescent pigment at 375 nm/430 nm.

Supplementary information

Supplementary Information

Supplementary Figs. 1–21.

Reporting Summary

Supplementary Video 1

Intracardial resin perfusion of a postnatal (P10) wildtype mouse. This video shows the intracardial resin perfusion of a postnatal (P10) wildtype mouse pub. All steps are precisely explained in the Procedure.

Supplementary Video 2

Three-dimensional animation of a reconstructed SRμCT scan of the entire vascular brain corrosion cast of an adult (P60) wildtype mouse. This animation shows a reconstructed SRμCT of an entire brain vascular corrosion cast of an adult (P60) wildtype animal. Five anatomical regions (cortex, hippocampus, superior colliculus, thalamus and cerebellum) were zoomed into with high resolution. Artificial coloring illustrates the vessel radius and the division between capillaries (vessel diameter <7 μm) and noncapillaries (vessel diameter ≥7 μm).

Fabrication of angstrom-scale two-dimensional channels for mass transport

Efficient and strand-specific profiling of replicating chromatin with enrichment and sequencing of protein-associated nascent DNA in mammalian cells

The eINTACT method for studying nuclear changes in host plant cells targeted by bacterial effectors in native infection contexts

Rapid reaction optimization by robust and economical quantitative benchtop19F NMR spectroscopy

Profiling native pulmonary basement membrane stiffness using atomic force microscopy

Photoimmunotechnology as a powerful biological tool for molecular-based elimination of target cells and microbes, including bacteria, fungi and viruses

Measuring key human carbohydrate digestive enzyme activities using high-performance anion-exchange chromatography with pulsed amperometric detection

Neural cell isolation from adult macaques for high-throughput analyses and neurosphere cultures

Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds

Neuronal subtype-specific growth cone and soma purification from mammalian CNS via fractionation and fluorescent sorting for subcellular analyses and spatial mapping of local transcriptomes and proteomes

查看全部

Sections

Figures

References

The formation of new blood vessels and the establishment of vascular networks are crucial during brain development, in the adult healthy brain, as well as in various diseases of the central nervous system. Here, we describe a step-by-step protocol for our
Introduction
Materials
Procedure
Troubleshooting
Timing
Anticipated results
References
ACKNOWLEDGMENTS
ACKNOWLEDGMENTS

Figure 1
Fig. 1: Flow chart, summary and time frame of the procedure.
Figure 2
Fig. 2: Intracardial resin perfusion and brain dissection for postnatal day 10 (P10) animals.
Figure 3
Fig. 3: The principles of (brain) vascular corrosion casting.
Figure 4
Fig. 4: The placement of the ROIs and process of computational 3D reconstruction.
Figure 5
$Fig. 5: Global vascular network morphometry: increased vascular volume fraction in various regions of the adult (P60) WT versus postnatal (P10) WT mouse brain.$ Fig. 5: Global vascular network morphometry: increased vascular volume fraction in various regions of the adult (P60) WT versus postnatal (P10) WT mouse brain.
Figure 6
Fig. 6: Distinguishing between capillaries and noncapillaries.
Figure 7
$Fig. 7: Local vascular network topology: increased vascular volume fraction of the adult (P60) WT versus postnatal (P10) WT cortex is mainly found at the capillary level.$ Fig. 7: Local vascular network topology: increased vascular volume fraction of the adult (P60) WT versus postnatal (P10) WT cortex is mainly found at the capillary level.
Figure 8
Fig. 8: Local vascular network topology: visualization and quantification of vascular branch point diameter and vascular branch point density in various regions of the adult (P60) WT versus the postnatal (P10) WT mouse brain.
Figure 9
Fig. 9: Local vascular network topology: segment density, diameter and length of the adult (P60) WT versus the postnatal (P10) WT mouse brain.
Figure 10
Fig. 10: Local vascular network topology: segment tortuosity and segment volume of the adult (P60) WT versus the postnatal (P10) WT mouse brain.
Figure 11
Fig. 11: Local vascular network topology: extravascular distance is decreased in various regions of the adult (P60) WT versus the postnatal (P10) WT mouse brain.
Figure 12
Fig. 12: Global vascular network morphology: visualization of distinct patterns of vessel directionality in various brain regions of the adult (P60) WT and postnatal (P10) WT mouse brain.
Figure 13
$Fig. 13: High-resolution SRµCT scan of the entire postnatal (P10) and adult (P60) mouse brain: visualization and quantification of various anatomical brain regions reveals increased vascular volume fractions in the P60 versus P10 mouse brain.$ Fig. 13: High-resolution SRµCT scan of the entire postnatal (P10) and adult (P60) mouse brain: visualization and quantification of various anatomical brain regions reveals increased vascular volume fractions in the P60 versus P10 mouse brain.
Figure 14
Fig. 14: Working model summarizing the differences in 3D vascular network architecture between adult (P60) WT and postnatal (P10) WT mice.

Yan, M. et al. Chronic DLL4 blockade induces vascular neoplasms. Nature 463, E6–E7 (2010).
Carmeliet, P. & Jain, R. K. Molecular mechanisms and clinical applications of angiogenesis. Nature 473, 298–307 (2011).
Potente, M., Gerhardt, H. & Carmeliet, P. Basic and therapeutic aspects of angiogenesis. Cell 146, 873–887 (2011).
Wälchli, T. et al. Wiring the vascular network with neural cues: a CNS perspective. Neuron 87, 271–296 (2015).
Jain, R. K. & Carmeliet, P. SnapShot: tumor angiogenesis. Cell 149, 1408–1408.e1401 (2012).
Herbert, S. P. & Stainier, D. Y. Molecular control of endothelial cell behaviour during blood vessel morphogenesis. Nat. Rev. Mol. Cell Biol. 12, 551–564 (2011).
Quaegebeur, A., Lange, C. & Carmeliet, P. The neurovascular link in health and disease: molecular mechanisms and therapeutic implications. Neuron 71, 406–424 (2011).
Burri, P. H., Hlushchuk, R. & Djonov, V. Intussusceptive angiogenesis: Its emergence, its characteristics, and its significance. Dev. Dyn. 231, 474–488 (2004).
Makanya, A. N., Hlushchuk, R. & Djonov, V. G. Intussusceptive angiogenesis and its role in vascular morphogenesis, patterning, and remodeling. Angiogenesis 12, 113–123 (2009).
Garcia-Gomez, P. & Valiente, M. Vascular co-option in brain metastasis. Angiogenesis https://doi.org/10.1007/s10456-019-09693-x (2019).
Kuczynski, E. A., Vermeulen, P. B., Pezzella, F., Kerbel, R. S. & Reynolds, A. R. Vessel co-option in cancer. Nat. Rev. Clin. Oncol. 16, 469–493 (2019).
Angara, K., Borin, T. F. & Arbab, A. S. Vascular mimicry: a novel neovascularization mechanism driving anti-angiogenic therapy (AAT) resistance in glioblastoma. Transl. Oncol. 10, 650–660 (2017).
Fernández-Cortés, M., Delgado-Bellido, D. & Oliver, F. J. Vasculogenic mimicry: become an endothelial cell “but not so much”. Front. Oncol. 9, 803–803 (2019).
Ricci-Vitiani, L. et al. Tumour vascularization via endothelial differentiation of glioblastoma stem-like cells. Nature 468, 824–828 (2010).
Wang, R. et al. Glioblastoma stem-like cells give rise to tumour endothelium. Nature 468, 829–833 (2010).
Cheng, L. et al. Glioblastoma stem cells generate vascular pericytes to support vessel function and tumor growth. Cell 153, 139–152 (2013).
Zhou, W. et al. Targeting glioma stem cell-derived pericytes disrupts the blood-tumor barrier and improves chemotherapeutic efficacy. Cell Stem Cell 21, 591–603.e594 (2017).
Coelho-Santos, V. & Shih, A. Y. Postnatal development of cerebrovascular structure and the neurogliovascular unit. Wiley Interdiscip. Rev. Dev. Biol. 9, e363–e363 (2020).
Mink, J. W., Blumenschine, R. J. & Adams, D. B. Ratio of central nervous system to body metabolism in vertebrates: its constancy and functional basis. Am. J. Physiol. Regul. Integr. Comp. Physiol. 241, R203–R212 (1981).
Begley, D. J. & Brightman, M. W. Structural and functional aspects of the blood-brain barrier. Prog. Drug Res. 61, 39–78 (2003).
Zlokovic, B. V. & Apuzzo, M. L. Strategies to circumvent vascular barriers of the central nervous system. Neurosurgery 43, 877–878 (1998).
Cassot, F., Lauwers, F., Lorthois, S., Puwanarajah, P. & Duvernoy, H. Scaling laws for branching vessels of human cerebral cortex. Microcirculation 16, 331–344 (2009).
Tsai, P. S. et al. Correlations of neuronal and microvascular densities in murine cortex revealed by direct counting and colocalization of nuclei and vessels. J. Neurosci. 29, 14553–14570 (2009).
Blinder, P. et al. The cortical angiome: an interconnected vascular network with noncolumnar patterns of blood flow. Nat. Neurosci. 16, 889–897 (2013).
Shih, A. Y. et al. Robust and fragile aspects of cortical blood flow in relation to the underlying angioarchitecture. Microcirculation 22, 204–218 (2015).
Harb, R., Whiteus, C., Freitas, C. & Grutzendler, J. In vivo imaging of cerebral microvascular plasticity from birth to death. J. Cereb. Blood Flow. Metab. 33, 146–156 (2013).
Iadecola, C. The neurovascular unit coming of age: a journey through neurovascular coupling in health and disease. Neuron 96, 17–42 (2017).
Vallon, M., Chang, J., Zhang, H. & Kuo, C. J. Developmental and pathological angiogenesis in the central nervous system. Cell. Mol. Life Sci. 71, 3489–3506 (2014).
Wälchli, T. et al. Quantitative assessment of angiogenesis, perfused blood vessels and endothelial tip cells in the postnatal mouse brain. Nat. Protoc. 10, 53–74 (2015).
Weller, R. O., Sharp, M. M., Christodoulides, M., Carare, R. O. & Møllgård, K. The meninges as barriers and facilitators for the movement of fluid, cells and pathogens related to the rodent and human CNS. Acta Neuropathol. 135, 363–385 (2018).
Hogan, K. A., Ambler, C. A., Chapman, D. L. & Bautch, V. L. The neural tube patterns vessels developmentally using the VEGF signaling pathway. Development 131, 1503–1513 (2004).
Paredes, I., Himmels, P. & Ruiz de Almodovar, C. Neurovascular communication during CNS development. Dev. Cell 45, 10–32 (2018).
Mancuso, M. R., Kuhnert, F. & Kuo, C. J. Developmental angiogenesis of the central nervous system. Lymphat. Res. Biol. 6, 173–180 (2008).
Fantin, A., Vieira, J. M., Plein, A., Maden, C. H. & Ruhrberg, C. The embryonic mouse hindbrain as a qualitative and quantitative model for studying the molecular and cellular mechanisms of angiogenesis. Nat. Protoc. 8, 418–429 (2013).
Vasudevan, A., Long, J. E., Crandall, J. E., Rubenstein, J. L. & Bhide, P. G. Compartment-specific transcription factors orchestrate angiogenesis gradients in the embryonic brain. Nat. Neurosci. 11, 429–439 (2008).
Fantin, A. et al. NRP1 acts cell autonomously in endothelium to promote tip cell function during sprouting angiogenesis. Blood 121, 2352–2362 (2013).
Daneman, R., Zhou, L., Kebede, A. A. & Barres, B. A. Pericytes are required for blood-brain barrier integrity during embryogenesis. Nature 468, 562–566 (2010).
Zeller, K., Vogel, J. & Kuschinsky, W. Postnatal distribution of Glut1 glucose transporter and relative capillary density in blood-brain barrier structures and circumventricular organs during development. Brain Res. Dev. Brain Res. 91, 200–208 (1996).
Wälchli, T. et al. Nogo-A is a negative regulator of CNS angiogenesis. Proc. Natl Acad. Sci. USA 110, E1943–E1952 (2013).
Wälchli, T. et al. Nogo-A regulates vascular network architecture in the postnatal brain. J. Cereb. Blood Flow. Metab. 37, 614–631 (2017).
Whiteus, C., Freitas, C. & Grutzendler, J. Perturbed neural activity disrupts cerebral angiogenesis during a postnatal critical period. Nature 505, 407–411 (2014).
Sweeney, M. D., Ayyadurai, S. & Zlokovic, B. V. Pericytes of the neurovascular unit: key functions and signaling pathways. Nat. Neurosci. 19, 771–783 (2016).
Muoio, V., Persson, P. B. & Sendeski, M. M. The neurovascular unit—concept review. Acta Physiol. 210, 790–798 (2014).
Eichmann, A. & Thomas, J. L. Molecular parallels between neural and vascular development. Cold Spring Harb. Perspect. Med. 3, a006551 (2013).
Yu, P. et al. FGF-dependent metabolic control of vascular development. Nature 545, 224–228 (2017).
Li, X., Sun, X. & Carmeliet, P. Hallmarks of endothelial cell metabolism in health and disease. Cell Metab. 30, 414–433 (2019).
Andrae, J., Gallini, R. & Betsholtz, C. Role of platelet-derived growth factors in physiology and medicine. Genes Dev. 22, 1276–1312 (2008).
Jih, Y. J. et al. Distinct regulation of genes by bFGF and VEGF-A in endothelial cells. Angiogenesis 4, 313–321 (2001).
Dong, J. et al. Glioma stem cells involved in tumor tissue remodeling in a xenograft model. J. Neurosurg. 113, 249–260 (2010).
Lawler, J. Thrombospondin-1 as an endogenous inhibitor of angiogenesis and tumor growth. J. Cell Mol. Med. 6, 1–12 (2002).
Lawler, P. R. & Lawler, J. Molecular basis for the regulation of angiogenesis by thrombospondin-1 and -2. Cold Spring Harb. Perspect. Med. 2, a006627 (2012).
O’Reilly, M. S. et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell 88, 277–285 (1997).
Dhanabal, M. et al. Angioarrestin: an antiangiogenic protein with tumor-inhibiting properties. Cancer Res. 62, 3834–3841 (2002).
Lu, X. et al. The netrin receptor UNC5B mediates guidance events controlling morphogenesis of the vascular system. Nature 432, 179–186 (2004).
Sakurai, A., Doçi, C. L. & Gutkind, J. S. Semaphorin signaling in angiogenesis, lymphangiogenesis and cancer. Cell Res. 22, 23–32 (2012).
Eilken, H. M. & Adams, R. H. Dynamics of endothelial cell behavior in sprouting angiogenesis. Curr. Opin. Cell Biol. 22, 617–625 (2010).
Strilic, B. et al. The molecular basis of vascular lumen formation in the developing mouse aorta. Dev. Cell 17, 505–515 (2009).
Pitulescu, M. E. et al. Dll4 and Notch signalling couples sprouting angiogenesis and artery formation. Nat. Cell Biol. 19, 915–927 (2017).
Lammert, E. & Axnick, J. Vascular lumen formation. Cold Spring Harb. Perspect. Med. 2, a006619 (2012).
Tung, J. J., Tattersall, I. W. & Kitajewski, J. Tips, stalks, tubes: Notch-mediated cell fate determination and mechanisms of tubulogenesis during angiogenesis. Cold Spring Harb. Perspect. Med. 2, a006601 (2012).
Wacker, A. & Gerhardt, H. Endothelial development taking shape. Curr. Opin. Cell Biol. 23, 676–685 (2011).
Mazzone, M. et al. Heterozygous deficiency of PHD2 restores tumor oxygenation and inhibits metastasis via endothelial normalization. Cell 136, 839–851 (2009).
Dela Paz, N. G., Melchior, B. & Frangos, J. A. Shear stress induces Gα(q/11) activation independently of G protein-coupled receptor activation in endothelial cells. Am. J. Physiol. Cell Physiol. 312, C428–C437 (2017).
Takuwa, N. et al. Tumor-suppressive sphingosine-1-phosphate receptor-2 counteracting tumor-promoting sphingosine-1-phosphate receptor-1 and sphingosine kinase 1—Jekyll Hidden behind Hyde. Am. J. Cancer Res. 1, 460–481 (2011).
Mendelson, K., Evans, T. & Hla, T. Sphingosine 1-phosphate signalling. Development 141, 5–9 (2014).
Gaengel, K. et al. The sphingosine-1-phosphate receptor S1PR1 restricts sprouting angiogenesis by regulating the interplay between VE-cadherin and VEGFR2. Dev. Cell 23, 587–599 (2012).
Jin, Z. G. et al. Ligand-independent activation of vascular endothelial growth factor receptor 2 by fluid shear stress regulates activation of endothelial nitric oxide synthase. Circ. Res. 93, 354–363 (2003).
Nigro, P., Abe, J. & Berk, B. C. Flow shear stress and atherosclerosis: a matter of site specificity. Antioxid. Redox Signal 15, 1405–1414 (2011).
Baeyens, N. et al. Defective fluid shear stress mechanotransduction mediates hereditary hemorrhagic telangiectasia. J. Cell Biol. 214, 807–816 (2016).
Franco, C. A. et al. Non-canonical Wnt signalling modulates the endothelial shear stress flow sensor in vascular remodelling. eLife 5, e07727–e07727 (2016).
Pries, A. R. & Secomb, T. W. Modeling structural adaptation of microcirculation. Microcirculation 15, 753–764 (2008).
Pries, A. R., Secomb, T. W. & Gaehtgens, P. Structural adaptation and stability of microvascular networks: theory and simulations. Am. J. Physiol. 275, H349–H360 (1998).
Hjelmeland, A. B., Lathia, J. D., Sathornsumetee, S. & Rich, J. N. Twisted tango: brain tumor neurovascular interactions. Nat. Neurosci. 14, 1375–1381 (2011).
Carmeliet, P., De Smet, F., Loges, S. & Mazzone, M. Branching morphogenesis and antiangiogenesis candidates: tip cells lead the way. Nat. Rev. Clin. Oncol. 6, 315–326 (2009).
Frahm, K. A., Nash, C. P. & Tobet, S. A. Endocan immunoreactivity in the mouse brain: method for identifying nonfunctional blood vessels. J. Immunol. Methods 398–399, 27–32 (2013).
Geudens, I. & Gerhardt, H. Coordinating cell behaviour during blood vessel formation. Development 138, 4569–4583 (2011).
Blanco, R. & Gerhardt, H. VEGF and Notch in tip and stalk cell selection. Cold Spring Harb. Perspect. Med. 3, a006569 (2013).
Phng, L. K. & Gerhardt, H. Angiogenesis: a team effort coordinated by notch. Dev. Cell 16, 196–208 (2009).
Noguera-Troise, I. et al. Blockade of Dll4 inhibits tumour growth by promoting non-productive angiogenesis. Nature 444, 1032–1037 (2006).
Thurston, G., Noguera-Troise, I. & Yancopoulos, G. D. The Delta paradox: DLL4 blockade leads to more tumour vessels but less tumour growth. Nat. Rev. Cancer 7, 327–331 (2007).
Lobov, I. B. et al. Delta-like ligand 4 (Dll4) is induced by VEGF as a negative regulator of angiogenic sprouting. Proc. Natl Acad. Sci. USA. 104, 3219–3224 (2007).
Ridgway, J. et al. Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis. Nature 444, 1083–1087 (2006).
Gest, T. R. & Carron, M. A. Embryonic origin of the caudal mesenteric artery in the mouse. Anat. Rec. A Discov. Mol. Cell Evol. Biol. 271, 192–201 (2003).
Adamson, S. L. et al. Interactions between trophoblast cells and the maternal and fetal circulation in the mouse placenta. Dev. Biol. 250, 358–373 (2002).
Iwagaki, T., Suzuki, T. & Nakashima, T. Development and regression of cochlear blood vessels in fetal and newborn mice. Hear. Res. 145, 75–81 (2000).
Carraro, M. & Harrison, R. V. Degeneration of stria vascularis in age-related hearing loss; a corrosion cast study in a mouse model. Acta Otolaryngol. 136, 385–390 (2016).
Carraro, M., Park, A. H. & Harrison, R. V. Partial corrosion casting to assess cochlear vasculature in mouse models of presbycusis and CMV infection. Hear. Res. 332, 95–103 (2016).
Carraro, M. et al. Cytomegalovirus (CMV) infection causes degeneration of cochlear vasculature and hearing loss in a mouse model. J. Assoc. Res. Otolaryngol. 18, 263–273 (2017).
Hossler, F. E., Lametschwandtner, A., Kao, R. & Finsterbusch, F. Microvascular architecture of mouse urinary bladder described with vascular corrosion casting, light microscopy, SEM, and TEM. Microsc. Microanal. 19, 1428–1435 (2013).
Peão, M. N., Aguas, A. P., de Sá, C. M. & Grande, N. R. Neoformation of blood vessels in association with rat lung fibrosis induced by bleomycin. Anat. Rec. 238, 57–67 (1994).
Peáo, M. N., Aguas, A. P., de Sá, C. M. & Grande, N. R. Identification of vascular sphincters at the junction between alveolar capillaries and pulmonary venules of the mouse lung. Anat. Rec. 241, 383–390 (1995).
Ackermann, M. et al. Effects of nintedanib on the microvascular architecture in a lung fibrosis model. Angiogenesis 20, 359–372 (2017).
Gibney, B. C. et al. Structural and functional evidence for the scaffolding effect of alveolar blood vessels. Exp. Lung Res. 43, 337–346 (2017).
Dahl, E. The fine structure of intracerebral vessels. Z. Zellforsch. Mikrosk. Anat. 145, 577–586 (1973).
Levesque, M. J., Cornhill, J. F. & Nerem, R. M. Vascular casting. A new method for the study of the arterial endothelium. Atherosclerosis 34, 457–467 (1979).
Duvernoy, H. M., Delon, S. & Vannson, J. L. Cortical blood vessels of the human brain. Brain Res. Bull. 7, 519–579 (1981).
Reina-De La Torre, F., Rodriguez-Baeza, A. & Sahuquillo-Barris, J. Morphological characteristics and distribution pattern of the arterial vessels in human cerebral cortex: a scanning electron microscope study. Anat. Rec. 251, 87–96 (1998).
Zagórska-Swiezy, K., Litwin, J. A., Gorczyca, J., Pityński, K. & Miodoński, A. J. Arterial supply and venous drainage of the choroid plexus of the human lateral ventricle in the prenatal period as revealed by vascular corrosion casts and SEM. Folia Morphol. (Warsz.) 67, 209–213 (2008).
Heinzer, S. et al. Hierarchical microimaging for multiscale analysis of large vascular networks. Neuroimage 32, 626–636 (2006).
Krucker, T., Lang, A. & Meyer, E. P. New polyurethane-based material for vascular corrosion casting with improved physical and imaging characteristics. Microsc. Res. Tech. 69, 138–147 (2006).
Sangiorgi, S. et al. Arterial and microvascular supply of cerebral hemispheres in the nude mouse revealed using corrosion casting and scanning electron microscopy. J. Anat. 232, 739–746 (2018).
Quintana, D. D. et al. The cerebral angiome: high resolution MicroCT imaging of the whole brain cerebrovasculature in female and male mice. NeuroImage 202, 116109 (2019).
Heinzer, S. et al. Novel three-dimensional analysis tool for vascular trees indicates complete micro-networks, not single capillaries, as the angiogenic endpoint in mice overexpressing human VEGF(165) in the brain. NeuroImage 39, 1549–1558 (2008).
Krucker, T., Schuler, A., Meyer, E. P., Staufenbiel, M. & Beckmann, N. Magnetic resonance angiography and vascular corrosion casting as tools in biomedical research: application to transgenic mice modeling Alzheimer’s disease. Neurol. Res. 26, 507–516 (2004).
Meyer, E. P., Ulmann-Schuler, A., Staufenbiel, M. & Krucker, T. Altered morphology and 3D architecture of brain vasculature in a mouse model for Alzheimer’s disease. Proc. Natl Acad. Sci. USA. 105, 3587–3592 (2008).
Walker, E. J., Shen, F., Young, W. L. & Su, H. Cerebrovascular casting of the adult mouse for 3D imaging and morphological analysis. J. Vis. Exp. https://doi.org/10.3791/2958 (2011).
Walker, E. J. et al. Arteriovenous malformation in the adult mouse brain resembling the human disease. Ann. Neurol. 69, 954–962 (2011).
Risser, L. et al. From homogeneous to fractal normal and tumorous microvascular networks in the brain. J. Cereb. Blood Flow. Metab. 27, 293–303 (2007).
Sangiorgi, S. et al. Early-stage microvascular alterations of a new model of controlled cortical traumatic brain injury: 3D morphological analysis using scanning electron microscopy and corrosion casting. J. Neurosurg. 118, 763 (2013).
Ohtake, M., Morino, S., Kaidoh, T. & Inoué, T. Three-dimensional structural changes in cerebral microvessels after transient focal cerebral ischemia in rats: scanning electron microscopic study of corrosion casts. Neuropathology 24, 219–227 (2004).
Rodríguez-Baeza, A., Reina-de la Torre, F., Poca, A., Martí, M. & Garnacho, A. Morphological features in human cortical brain microvessels after head injury: a three-dimensional and immunocytochemical study. Anat. Rec. A Discov. Mol. Cell Evol. Biol. 273, 583–593 (2003).
Zhang, F., Wen, Y. & Guo, X. CRISPR/Cas9 for genome editing: progress, implications and challenges. Hum. Mol. Genet. 23, R40–R46 (2014).
Gerhardt, H. et al. Neuropilin-1 is required for endothelial tip cell guidance in the developing central nervous system. Dev. Dyn. 231, 503–509 (2004).
Gerhardt, H. et al. VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia. J. Cell Biol. 161, 1163–1177 (2003).
Provost, J. et al. 3-D ultrafast Doppler imaging applied to the noninvasive mapping of blood vessels in vivo. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 62, 1467–1472 (2015).
Demene, C. et al. 4D microvascular imaging based on ultrafast Doppler tomography. NeuroImage 127, 472–483 (2016).
Pathak, A. P., Kim, E., Zhang, J. & Jones, M. V. Three-dimensional imaging of the mouse neurovasculature with magnetic resonance microscopy. PLoS One 6, e22643–e22643 (2011).
Hartung, M. P., Grist, T. M. & François, C. J. Magnetic resonance angiography: current status and future directions. J. Cardiovasc. Magn. Reson. 13, 19–19 (2011).
Kelch, I. D. et al. Organ-wide 3D-imaging and topological analysis of the continuous microvascular network in a murine lymph node. Sci. Rep. 5, 16534 (2015).
Marien, K. M. et al. Development and validation of a histological method to measure microvessel density in whole-slide images of cancer tissue. PLoS One 11, e0161496–e0161496 (2016).
Bukenya, F., Nerissa, C., Serres, S., Pardon, M. C. & Bai, L. An automated method for segmentation and quantification of blood vessels in histology images. Microvasc. Res. 128, 103928 (2020).
Oren, R. et al. Whole organ blood and lymphatic vessels imaging (WOBLI). Sci. Rep. 8, 1412–1412 (2018).
Kennel, P., Teyssedre, L., Colombelli, J. & Plouraboué, F. Toward quantitative three-dimensional microvascular networks segmentation with multiview light-sheet fluorescence microscopy. J. Biomed. Opt. 23, 1–14 (2018).
Di Giovanna, A. P. et al. Tailored sample mounting for light-sheet fluorescence microscopy of clarified specimens by polydimethylsiloxane casting. Front. Neuroanat. 13, 35–35 (2019).
Lugo-Hernandez, E. et al. 3D visualization and quantification of microvessels in the whole ischemic mouse brain using solvent-based clearing and light sheet microscopy. J. Cereb. Blood Flow. Metab. 37, 3355–3367 (2017).
Todorov, M. I. et al. Machine learning analysis of whole mouse brain vasculature. Nat. Methods 17, 442–449 (2020).
Kirst, C. et al. Mapping the fine-scale organization and plasticity of the brain vasculature. Cell 180, 780–795.e725 (2020).
Susaki, E. A. & Ueda, H. R. Whole-body and whole-organ clearing and imaging techniques with single-cell resolution: toward organism-level systems biology in mammals. Cell Chem. Biol. 23, 137–157 (2016).
Zagorchev, L. et al. Micro computed tomography for vascular exploration. J. Angiogenes. Res. 2, 7 (2010).
Blery, P. et al. Vascular imaging with contrast agent in hard and soft tissues using microcomputed-tomography. J. Microsc. 262, 40–49 (2016).
Starosolski, Z. et al. Ultra high-resolution in vivo computed tomography imaging of mouse cerebrovasculature using a long circulating blood pool contrast agent. Sci. Rep. 5, 10178 (2015).
Beckmann, N. et al. Age-dependent cerebrovascular abnormalities and blood flow disturbances in APP23 mice modeling Alzheimer’s disease. J. Neurosci. 23, 8453–8459 (2003).
Gore, A. V., Monzo, K., Cha, Y. R., Pan, W. & Weinstein, B. M. Vascular development in the zebrafish. Cold Spring Harb. Perspect. Med. 2, a006684 (2012).
Dorr, A., Sled, J. G. & Kabani, N. Three-dimensional cerebral vasculature of the CBA mouse brain: a magnetic resonance imaging and micro computed tomography study. NeuroImage 35, 1409–1423 (2007).
Figueiredo, G., Boll, H., Kramer, M., Groden, C. & Brockmann, M. A. In vivo X-ray digital subtraction and CT angiography of the murine cerebrovasculature using an intra-arterial route of contrast injection. Am. J. Neuroradiol. 33, 1702 (2012).
Ben-Zvi, A. et al. Mfsd2a is critical for the formation and function of the blood-brain barrier. Nature 509, 507–511 (2014).
Vanlandewijck, M. et al. A molecular atlas of cell types and zonation in the brain vasculature. Nature 554, 475–480 (2018).
Kalucka, J. et al. Single-cell transcriptome atlas of murine endothelial cells. Cell 180, 764–779.e720 (2020).
Goveia, J. et al. An integrated gene expression landscape profiling approach to identify lung tumor endothelial cell heterogeneity and angiogenic candidates. Cancer Cell 37, 21–36.e13 (2020).
Pitulescu, M. E., Schmidt, I., Benedito, R. & Adams, R. H. Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice. Nat. Protoc. 5, 1518–1534 (2010).
Sawamiphak, S., Ritter, M. & Acker-Palmer, A. Preparation of retinal explant cultures to study ex vivo tip endothelial cell responses. Nat. Protoc. 5, 1659–1665 (2010).
Willner, M. et al. Quantitative three-dimensional imaging of lipid, protein, and water contents via X-ray phase-contrast tomography. PLoS One 11, e0151889 (2016).
Moyon, D., Pardanaud, L., Yuan, L., Bréant, C. & Eichmann, A. Plasticity of endothelial cells during arterial-venous differentiation in the avian embryo. Development 128, 3359–3370 (2001).
Villa, N. et al. Vascular expression of Notch pathway receptors and ligands is restricted to arterial vessels. Mech. Dev. 108, 161–164 (2001).
Seki, T., Yun, J. & Oh, S. P. Arterial endothelium-specific activin receptor-like kinase 1 expression suggests its role in arterialization and vascular remodeling. Circ. Res. 93, 682–689 (2003).
Cui, X. et al. Venous endothelial marker COUP-TFII regulates the distinct pathologic potentials of adult arteries and veins. Sci. Rep. 5, 16193 (2015).
You, L. R. et al. Suppression of Notch signalling by the COUP-TFII transcription factor regulates vein identity. Nature 435, 98–104 (2005).
Wang, H. U., Chen, Z. F. & Anderson, D. J. Molecular distinction and angiogenic interaction between embryonic arteries and veins revealed by ephrin-B2 and its receptor Eph-B4. Cell 93, 741–753 (1998).
Malcontenti-Wilson, C., Chan, L., Nikfarjam, M., Muralidharan, V. & Christophi, C. Vascular targeting agent Oxi4503 inhibits tumor growth in a colorectal liver metastases model. J. Gastroenterol. Hepatol. 23, e96–e104 (2008).
Kaidoh, T., Yasugi, T. & Uehara, Y. The microvasculature of the 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumour. Virchows Arch. A 418, 111–117 (1991).
Chakhoyan, A. et al. Validation of vessel size imaging (VSI) in high-grade human gliomas using magnetic resonance imaging, image-guided biopsies, and quantitative immunohistochemistry. Sci. Rep. 9, 2846 (2019).
Burrell, J. S. et al. MRI measurements of vessel calibre in tumour xenografts: Comparison with vascular corrosion casting. Microvasc. Res. 84, 323–329 (2012).
O’Connor, J. P. et al. Quantifying antivascular effects of monoclonal antibodies to vascular endothelial growth factor: insights from imaging. Clin. Cancer Res. 15, 6674–6682 (2009).
Miyake, T., Murakami, T. & Ohtsuka, A. Incomplete vascular casting for a scanning electron microscope study of the microcirculatory patterns in the rat pancreas. Arch. Histol. Cytol. 55, 397–406 (1992).
Christoffersonm, R. H. & Nilsson, B. O. Microvascular corrosion casting with analysis in the scanning electron microscope. Scanning 10, 43–63 (1988).
Paganin, D., Mayo, S. C., Gureyev, T. E., Miller, P. R. & Wilkins, S. W. Simultaneous phase and amplitude extraction from a single defocused image of a homogeneous object. J. Microsc. 206, 33–40 (2002).
Marone, F. & Stampanoni, M. Regridding reconstruction algorithm for real-time tomographic imaging. J. Synchrotron Radiat. 19, 1029–1037 (2012).
Miettinen, A., Oikonomidis, I. V., Bonnin, A. & Stampanoni, M. NRStitcher: non-rigid stitching of terapixel-scale volumetric images. Bioinformatics 35, 5290–5297 (2019).
Otsu, N. A threshold selection method from gray-level histograms. IEEE Trans. Syst. Man Cybern. 9, 62–66 (1979).
Huang, L.-K. & Wang, M.-J. J. Image thresholding by minimizing the measures of fuzziness. Pattern Recogn. 28, 41–51 (1995).
Prewitt, J. M. S. & Mendelsohn, M. L. The analysis of cell images. Ann. NY Acad. Sci. 128, 1035–1053 (1966).
Li, C. H. & Tam, P. K. S. An iterative algorithm for minimum cross entropy thresholding. Pattern Recogn. Lett. 19, 771–776 (1998).
Lee, T. C., Kashyap, R. L. & Chu, C. N. Building skeleton models via 3-D medial surface axis thinning algorithms. CVGIP Graph. Models Image Process. 56, 462–478 (1994).
Calvin, R., Maurer, J., Qi, R. & Raghavan, V. A linear time algorithm for computing exact euclidean distance transforms of binary images in arbitrary dimensions. IEEE Trans. Pattern Anal. Mach. Intell. 25, 265–270 (2003).
Suhadolnik, A., Petrišič, J. & Kosel, F. An anchored discrete convolution algorithm for measuring length in digital images. Measurement 42, 1112–1117 (2009).
Tukey, J. W. Exploratory Data Analysis (Addison-Wesley, 1977).
ACSF for LSPS. Cold Spring Harbor Protocols https://doi.org/10.1101/pdb.rec071944 (2012).
Heinzer, S. et al. Computer-based analysis of microvascular alterations in a mouse model for Alzheimer’s disease. Proc. SPIE 6511, 65114 (2007).
Rust, R. et al. Nogo-A targeted therapy promotes vascular repair and functional recovery following stroke. Proc. Natl Acad. Sci. 116, 14270 (2019).
Joly, S., Dejda, A., Rodriguez, L., Sapieha, P. & Pernet, V. Nogo-A inhibits vascular regeneration in ischemic retinopathy. Glia 66, 2079–2093 (2018).
Seiler, S., Di Santo, S. & Widmer, H. R. Nogo-A neutralization improves graft function in a rat model of Parkinson’s disease. Front. Cell Neurosci. 10, 87 (2016).
Weber, B., Keller, A. L., Reichold, J. & Logothetis, N. K. The microvascular system of the striate and extrastriate visual cortex of the macaque. Cereb. Cortex 18, 2318–2330 (2008).
Fenrich, K. K., Zhao, E. Y., Wei, Y., Garg, A. & Rose, P. K. Isolating specific cell and tissue compartments from 3D images for quantitative regional distribution analysis using novel computer algorithms. J. Neurosci. Methods 226, 42–56 (2014).

Hierarchical imaging and computational analysis of three-dimensional vascular network architecture in the entire postnatal and adult mouse brain

Extended

Supplementary information

推荐阅读