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Skinned nerve fiber was prepared by treating a giant axon of squid with saponin. Filamentous networks and membranous organelles such as vacuoles, smooth membranes and mitochondria were present after the extraction. Ruthenium red staining of the extracted tissue showed dense fibrillar networks in the cytoplasm that correspond to the "microtrabeculae" or "cytoskeleton" reported by other investigators. Our method of preparation, thus, gives further evidence that these structures are not the result of artifactual condensation or precipitation. The present results also indicated that the networks are composed of acid mucopolysaccharides and/or mucoproteins. The density of microtubules was not changed by the extraction per se, but the number of microtubules remaining after extraction decreased by alteration of environmental conditions such as low temperature (4 degrees C), or the presence of Ca++ (1 mM), Mg++ free medium or colchicine (1 mM). These conditions are known to inhibit axonal transport. The usefulness of our extracted cytoplasm as a model of microtubule-related physiological functions is discussed.