Abstract
Numerous small vessels making up the central nervous system blood and lymphatic vascular networks are
heterogeneous and region-specific dynamic structures, whose segments, position, shape and function can
change in response to physiological and pathophysiological conditions. To date it has not been possible to
integrate blood and lymphatic vascular elements and their microenvironment to achieve a holistic
quantitative characterization of the combined brain and meningeal tissue-scale vascular networks, its
structure and function in normal and disease states. This application proposes to develop microscopy-
based high-throughput image analysis techniques for automated extraction of blood and lymphatic vascular
networks enabling quantitative morphodynamic characterization of cerebrovascular microenvironment
changes in two intracranial compartments – the brain and dura mater. The study will focus on new
algorithms for precise region-specific microvessel registration, mosaicing, segmentation, fusion and
colocalization for constructing large tissue scale spatially aligned dual blood/lymphatic vascular network
structural maps in the animals of both sexes, as well as characterization of heterogeneities of microvascular
networks, including blood and lymphatic vasculature, under estrogen and sleep deprivation (the conditions
relevant to multiple cerebrovascular disorders) compared to physiological settings. In other words,
advanced microscopy-based techniques will be used to image blood and lymphatic vessels at sub-micron
resolution in dura mater and the brain, and then cutting-edge deep machine learning imaging analysis
methods will be employed to segment and quantify these vessels, their geometry, vessel wall structure,
functionality, and interrelationship. Detailed structural analysis of microvascular networks is essential for
accurate evaluation of the distribution of physical forces, substrate delivery and tissue clearance of waste,
as well as sex differences and consequences of intracranial networks remodeling under physiological and
pathological conditions. This will create knowledge enabling a better understanding of the pathogenesis of
vascular impairments under estrogen and sleep deprivation, identify common molecular mechanisms and
the efficacy and effectiveness of different therapeutic treatments. Without the ability to construct total
structural and functional blood/lymphatic vascular network maps from studies limited to individual tissue
component parts, it is little wonder that translation from the molecular and cellular levels to the whole organ
and system levels is deficient and hinders translational progress towards a comprehensive understanding of
the pathophysiology associated with a range of neurological disorders.
