Measurement of solute permeability in the mouse spinal cord.

BACKGROUND: Sensory perception and motor dexterity is coordinated by the spinal cord, which remains effective due to maintenance of neuronal homeostasis. This is stringently controlled by the blood spinal cord barrier. Therefore, the function of the spinal cord is susceptible to alterations in the microvessel integrity (e.g. vascular leakage) and/or perfusion (e.g. changes in blood flow).

NEW METHOD: Spinal cord solute permeability was measured in anaesthetised mice. The lumbar spinal cord vertebra were stabilised and a coverslip secured to allow fluorescent tracers of vascular function and anatomy to be visualised in the vascular network. Fluorescence microscopy allowed real time measurements of vascular leakage and capillary perfusion within the spinal cord.

RESULTS: Capillaries were identified through fluorescent labelling of the endothelial luminal glycocalyx (wheat germ agglutin 555). Real time estimation of vascular permeability through visualisation of sodium fluorescein transport was recorded from identified microvessels in the lumbar dorsal horn of the spinal cord.

COMPARISON WITH EXISTING METHOD(S): Current approaches have used histological and/or tracer based in-vivo assays alongside cell culture to determine endothelium integrity and/or function. These only provide a snapshot of the developing vasculopathy, restricting the understanding of physiological function or disease progression over time.

CONCLUSIONS: These techniques allow for direct visualisation of cellular and/or mechanistic influences upon vascular function and integrity, which can be applied to rodent models including disease, transgenic and/or viral approaches. This combination of attributes allows for real time understanding of the function of the vascular network within the spinal cord.