Cam Y, Ledig M, Ebel A, Sensenbrenner M, Mandel P
Neurochem Res. 1980 Aug;5(8):831-45. doi: 10.1007/BF00965783.
Brain extracts from 8-day-old chick embryos have been shown to influence morphological development of dissociated brain cells from 7-day-old chick embryos in culture. Stimulatory effects on size of the neuronal somas and on growth of long processes were observed by adding the cytosol of the brain extract or the dialysate of the cytosol. These morphological changes parallel modifications of various enzyme activities according to the age of the cultures. Adenyl cyclase, (Na+,K+)- and Mg2+-ATPase, 5'-nucleotidase, choline acetyltransferase, and acetylcholinesterase activities were studied between 5 and 14 days of culture. Adenyl cyclase activity was strongly stimulated at 8 days by both extracts. (Na+,K+)- and Mg2+-ATPase activities were stimulated in 8-day-old cultures only by the dialysate. 5'-Nucleotidase activity was stimulated in 8-day-old cultures by the dialysate and in 11-day-old cultures by both extracts. Choline acetyltransferase activity was stimulated by the cytosol in 8-day-old cultures and by the dialysate in 11-day-old cultures. The total acetylcholinesterase activity was higher in 8-, 11-, and 14-day-old cultures treated with the cytosol. When the cells were treated with the dialysate, the activity was only higher in 14-day-old cultures. We also found that following the addition of brain extracts, the specific activity of the enzymes we studied was enhanced and became close to the values found in vivo during embryogenesis. Thus in parallel to the morphological modifications observed in nerve cell cultures treated by embryo brain extracts, biochemical variations especially involved in synaptogenesis and membrane development could be measured.
已证明,8日龄鸡胚的脑提取物会影响体外培养的7日龄鸡胚解离脑细胞的形态发育。通过添加脑提取物的胞质溶胶或胞质溶胶的透析液,观察到对神经元胞体大小和长突起生长的刺激作用。这些形态变化与培养物不同年龄时各种酶活性的改变平行。在培养的第5至14天研究了腺苷酸环化酶、(钠+,钾+)-和镁2+-ATP酶、5'-核苷酸酶、胆碱乙酰转移酶和乙酰胆碱酯酶的活性。两种提取物在第8天均强烈刺激腺苷酸环化酶活性。仅透析液在8日龄培养物中刺激(钠+,钾+)-和镁2+-ATP酶活性。透析液在8日龄培养物中刺激5'-核苷酸酶活性,两种提取物在11日龄培养物中刺激该酶活性。胆碱乙酰转移酶活性在8日龄培养物中受胞质溶胶刺激,在11日龄培养物中受透析液刺激。用胞质溶胶处理的8、11和14日龄培养物中总乙酰胆碱酯酶活性较高。当细胞用透析液处理时,该活性仅在14日龄培养物中较高。我们还发现,添加脑提取物后,我们研究的酶的比活性增强,并接近胚胎发育过程中体内发现的值。因此,与胚胎脑提取物处理的神经细胞培养物中观察到的形态学改变平行,可以测量特别是参与突触形成和膜发育的生化变化。