Differences between the peripheral and the central nervous system in permeability to sodium fluorescein

Sodium fluorescein (SF) was used as a very small tracer (mol wt 376; 5 A diameter) to examine diffusion barriers in peripheral nerves and to compare them to those in other regions of the nervous system. The technique involved immobilization of the tracer by rapid freezing, followed by freeze-drying and vacuum embedding in paraffin. The localization of the SF was then determined in tissue secretions using fluorescence microscopy. Even at the highest doses of intravenously (IV) injected tracer, no extravasation could be detected in the cerebral cortex. On the other hand, SF penetrated very rapidly into peripheral ganglia and into the epineurium and perineurium of large peripheral nerves. The penetration of SF into the endoneurium of large nerves was, however, much more restricted with tracer detectable within the endoneurium only at high doses and long survival times. Even in such cases, the level of SF fluorescence was much lower within nerve fascicles than in the epineurium and the perineurium, and a sharp gradient in fluorescence intensity persisted at the inner border of the perineurium. The extent of extravasation into the endoneurium varied markedly betwen different fascicles of the same nerve and between different nerves in the same animal. Experiments involving injection of high doses of SF adjacent to the nerve indicated relatively little movement of SF across the perineurium, which indicates that the observed accumulation of tracer within the endoneurium was the result of direct extravasation of SF from the endoneural blood vessels. Small nerve branches (< 100 mu diameter) showed an earlier and more extensive penetration of SF into the endoneurium than large nerves like the sciatic, hypoglossal, or ventral tail nerve. This may be due to a diffusion of SF along the extracellular space of the endoneurium from nerve terminals where the perineurial barrier is open-ended. In experiments involving IV injection of a solution containing both green fluorescent SF and red fluorescent Evans Blue (Evans Blue-serum albumin conplex, EBA = mol wt. 69,000), the distribution of SF could be directly compared at various sites and sacrifice times to that of EBA, a much larger tracer. SF appeared more rapidly and extensively than EBA in the various compartments in ganglia and peripheral nerve. The distribution of EBA was the same as is typically seen when this tracer is injected alone, indicating that there was no change in vascular permeability associated with IV injection of SF. Since SF is of very small size, freely diffusible, nontoxic, and detectable at very low concentrations, it should be a useful complement to existing tracers. When tissues are processed according to the indicated procedure, one can obtain a very sensitive and reliable localization of this tracer which should be of value for studies in the nervous system concerning various pathological conditions associated with permeability alterations.