Ionic permeability of the frog sciatic nerve perineurium: parallel studies of potassium and lanthanum penetration using electrophysiological and electron microscopic techniques

Abstract

The isolated sciatic nerve of the frog Rana temporaria was used for a parallel electrophysiological and electron microscopic examination of the ionic permeability of the perineurium, one component of the blood-nerve barrier. Nerves mounted in a grease-gap chamber for electrophysiological recording showed negligible changes in DC potential (Delta DC) or compound action potential on challenge with 100 mM K(+) Ringer, evidence that the perineurium was tight to K(+). In preparations then fixed and exposed to 5 mM lanthanum in the fixative, and examined in the electron microscope, electron-dense lanthanum deposits were seen between perineurial lamellae, but lanthanum was not detectable within the endoneurium, confirming that the perineurium was also tight to lanthanum. Absence of lanthanum penetration was confirmed by X-ray analysis of electron microscopic sections. In nerves exposed to 2 mM sodium deoxycholate (DOC) in the recording chamber, then challenged with high [K(+)], a moderate increase in perineurial K(+) permeability (P(K)) was observed, but lanthanum was still excluded. Exposure of nerves to 4 mM DOC caused a greater increase in perineurial potassium permeability, and the two nerves with the greatest permeability (P(K) > 1 x 10(-5) cm x sec(-1)) also showed detectable lanthanum within the endoneurium. The results indicate that DOC causes a dose-dependent increase in tight junctional permeability in the perineurium, and that the electrophysiological monitoring of K(+) penetration is a more sensitive measure of small ion permeability than electron microscopical analysis using lanthanum as tracer. Vesicular profiles observed in perineurial lamellae did not form open channels for ion flux across the perineurium in control nerves, or in those exposed to DOC. In preparations where lanthanum reached the endoneurium, lanthanum was observed in dense deposits in the extracellular spaces around nodes of Ranvier, and in the outer mesaxon cleft, but did not penetrate the internodal periaxonal space, the myelin intraperiod line, or the Schmidt-Lanterman incisures, in contrast to observations in mammalian nerves. The apparent differences in accessibility of the internodal periaxonal space in frog and mammalian axons are discussed in relation to axonal physiology. The study illustrates the value of parallel electrophysiological and electron microscopic examination in elucidating the properties of extracellular ionic pathways and their role in neural function.