Characterization of Neuronal and Glial Fractions Separated in Sucrose and Ficoll Media

摘要: Rat brain cerebral cortices were dispersed by a collagenase-hyaluronidase treatment and the suspensions obtained fractionated in sucrose Ficoll systems centrifugation. For short sedimentation run was followed density gradient; for only gradient used. Isolated cell populations examined light microscopy differential counts taken. Separation yielded more homogenous but smaller fractions. In up to 37 million neuronal glial cells could be collected from 5 cortices. Cells isolated contained 70 some 50 pg protein, respectively. Neuronal separated accumulated potassium, metabolically active. Both respiration incorporation of radioactive leucine into protein clearly active Krebs-Ringer phosphate buffer containing 10 mM glucose, neurons respired 946 875, glia 706 396 nmoles oxygen/mg protlhr after separation respectively. Incorporation 5370 4980 dpm/mg prot neurons, 3110 4480 glia. On other hand, no clear difference found TCA-soluble radioactivities tween