Biochemical and Ultrastructural Studies of Cultured Rat Astroglial Cells: Effect of Brain Extract and Dibutyryl Cyclic AMP on Glial Fibrillary Acidic Protein and Glial Filaments

Abstract

The growth of astroglial cells in primary cultures derived from newborn rat cerebral hemispheres was investigated in the absence and in the presence of newborn rat brain extract or dBcAMP. The parameters chosen were the content of DNA, total protein, and glial fibrillary acidic protein (GFA) as well as the morphologic development of gliofilaments. During the entire culture period the DNA content increased in control culture indicating a continuous cell division, whereas the cells stopped dividing after 14 or 4 days of treatment with either brain extract or dBcAMP respectively. In contrast, a constant increase of total protein was found in both control and treated cultures. Since cell divisions had stopped in treated cultures, the increase in total protein in these cultures indicates growth of the individual cells. The GFA levels increased progressively and similarly in control cultures and in cultures treated with brain extract. The values in the treated cultures remained slightly higher than those in controls. Conversely, immediately after the addition of dBcAMP a sudden increase in GFA protein occurred and the amounts were statistically significantly different from those of the controls. The GFA levels were expressed relative to total protein indicating that GFA constitutes an increasing amount of the total protein of the individual cells during culture. The changes in the amount of GFA was shown to parallel the morphologic development of gliofilaments. Indeed, when the level of GFA increased a progressive accumulation of gliofilaments was observed. The results obtained were discussed in relation to the astrocytic maturation.