Conditions affecting the extracellular space in the frog's forebrain

Abstract
The forebrain of the isolated central nervous system of frogs was fixed by freeze substitution and studied with the electron microscope. The extracellular space (ECS) of brains kept for 15–20 min in a physiological salt solution at room temperature varied from an appreciable to a negligible one. In electron micrographs exhibiting a large ECS the tissue elements had a uniform electron density. The EMs with little space featured in some instances a moderate swelling of presynaptic terminals and other tissue elements. Brains kept in a cooled medium or a salt solution with MgCl2 added exhibited invariably an abundant ECS. Treating the brain with a 100 mM KCl solution 5–90 sec before freezing yielded EMs with a contracted ECS and swollen tissue elements, many of which could be identified as dendritic spines. This effect of KCl was in many experiments prevented by bathing the brain in a salt solution containing 10 mM MgCl2 or in a Ca‐free solution.