Dissociated cells from 5- to 12-day-old chick embryo cerebral hemispheres were cultivated in polylysine-coated plastic Petri dishes. The polylysine substrate was observed to be favorable for the growth of neuronal cells, whereas glioblast proliferation was inhibited. The optimal conditions for the production of a predominantly neuronal culture were to use cerebral hemispheres from 7-day-old chick embryos, to dissociate the brain tissue mechanically and to seed the cells at a concentration range between 1.5 and 5 × 106 cells/ml. The cultures were observed by phase contrast microscopy. Most cells grew fibers and differentiated into bipolar and multipolar neurons. These neurons were stained by thionine, which demonstrated the presence of Nissl bodies. The silver impregnation revealed the presence of neurofibrils within the nerve fibers. Acetylcholinesterase was found to be present in the neuronal cells, but absent in the glioblasts. Under our culture conditions the neurons survived for 10–12 days. This system should allow further studies on the effects of growth factors on the differentiation of isolated neurons as well as investigations on neuron-glial interrelationship.
This study evaluates the hypothesis that arginine vasopressin (AVP) and atriopeptin, peptide hormones synthesized and released within the brain, are regulators of brain cell volume using cultured astroglial cells derived from newborn rats. Cell water content, regarded as volume, was measured in defined, serum-free medium as the 3-O-methylglucose (3-MG) space. Initial experiments established conditions such that glucose, which competes with 3-MG for the glucose carrier, would not interfere with the measurement of the 3-MG space. AVP increased the 3-MG space of glial cells by an average of 25% between 30 and 120 min of exposure, whereas atriopeptin decreased it by 32%. The 3-MG space remained close to normal after coadministration of both peptides. The AVP-dependent increase in 3-MG space was blocked both by the V1 antagonist d(CH2)5Tyr(Me)AVP (Manning compound) and by the cotransport inhibitor, bumetanide. Results are consistent with a role for AVP and atriopeptin in the homeostasis of atroglial cell volume.
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