Shambaugh G E, Koehler R R, Radosevich J A
VA Lakeside Medical Center, Chicago, Illinois.
Exp Cell Res. 1987 Oct;172(2):404-16. doi: 10.1016/0014-4827(87)90398-3.
The glycolytic enzyme enolase increases during the perinatal period of brain development and was utilized as a marker for examining the effect of culture environment on differentiation of cells from 20-day fetal rat brain. Enolase activity in cell cultures increased from 0.91 +/- 0.03 (Day 0) to 2.11 +/- 0.10 mumol/min/mg protein (Day 6). Comparable levels were not reached in vivo until neonatal pups were 15 days old. The in vitro increase was inhibited by both cycloheximide and actinomycin D. Enolase activity in the cells responded to alterations in both incubation media and homologous serum. After 6 days in culture, cells incubated in rat serum (10%) added to MEM or RPMI produced twice as much enolase activity as cells incubated similarly in Ham's medium, i.e., 1.96 +/- 0.09 and 1.85 +/- 0.21 vs 1.02 +/- 0.09, P less than 0.001. Results of a comparable magnitude were obtained when fetal calf serum replaced adult rat serum, but enolase production was somewhat lower when newborn calf serum replaced adult rat or fetal calf serum. When cells were incubated for 6 days with graded concentrations of adult rat serum (2.5-15%), enolase activity increased progressively. The pattern of enolase response suggests that the fetal rat brain cell model described herein will provide a sensitive probe with which to gain insight into nutrition and fetal brain development.
糖酵解酶烯醇化酶在脑发育的围产期增加,并被用作检测培养环境对20日龄胎鼠脑细胞分化影响的标志物。细胞培养物中的烯醇化酶活性从0.91±0.03(第0天)增加到2.11±0.10μmol/分钟/毫克蛋白质(第6天)。在体内,直到新生幼崽15日龄时才达到可比水平。体外增加受到放线菌酮和放线菌素D的抑制。细胞中的烯醇化酶活性对孵育培养基和同源血清的变化均有反应。在培养6天后,添加到MEM或RPMI中的10%大鼠血清中孵育的细胞产生的烯醇化酶活性是在Ham培养基中类似孵育细胞的两倍,即1.96±0.09和1.85±0.21对比1.02±0.09,P<0.001。当胎牛血清替代成年大鼠血清时获得了类似程度的结果,但当新生牛血清替代成年大鼠或胎牛血清时,烯醇化酶的产生略低。当细胞在不同浓度(2.5-15%)的成年大鼠血清中孵育6天时,烯醇化酶活性逐渐增加。烯醇化酶的反应模式表明,本文所述的胎鼠脑细胞模型将提供一个敏感的探针,用于深入了解营养与胎儿脑发育。