Echevarría D, Albus K
Department of Neurobiology/192, Max Planck Institute for Biophysical Chemistry, P.O. Box 2841, D-37070 Göttingen, Germany.
Brain Res Dev Brain Res. 2000 Oct 28;123(2):151-64. doi: 10.1016/s0165-3806(00)00089-4.
The development of spontaneous bioelectric activity (SBA) in organotypic tissue cultures (OTCs) from rat occipital cortex was studied by means of extracellular recording techniques in OTCs grown normally for 6-51 days in vitro (DIV), and in OTCs in which SBA had been silenced from DIV 4 on for 2 to 3 weeks by elevating the Mg(2+) levels in the growth medium. The proportions of spontaneously active neurones increased from about 25% at 6-14 DIV to more than 80% beyond the third week in vitro. Mature neurones discharged at shorter intervals and more vigorously than immature neurones; the developmental increase in firing rate was not significant, however. In OTCs 6-14 DIV the majority of spontaneously active neurones fired sluggishly in a regular manner. The remaining neurones fired action potentials in the form of discrete bursts resembling interictal activity in vivo. The proportions of these neurones increased from about 40% at 6-14 DIV to more than 80% beyond the third week in vitro. During development in vitro the mean burst duration increased from 3.5 s to about 8 s whereas the mean burst rate (between 0.7-1 bursts/min) remained constant. Activity-deprived neurons had low firing rates and fired action potentials in the form of discrete bursts with a mean burst rate of 0.4/min. The proportions of spontaneously active neurons, the variability of neuronal firing and the viability of the explants either were not altered by the activity blockade or had recovered to control values after 5-6 days in normal growth medium. We conclude that in OTCs of rat neocortex the absence of SBA during development in vitro delays the maturation of excitatory mechanisms responsible for the developmental increase in firing intensity. The development of burst firing modes is less affected by activity blockade.
利用细胞外记录技术,研究了大鼠枕叶皮质器官型组织培养物(OTCs)中自发生物电活动(SBA)的发育情况。这些OTCs在体外正常培养6 - 51天(体外培养天数,DIV),以及从第4天起通过提高生长培养基中镁离子(Mg(2+))水平使SBA沉默2至3周的OTCs。自发活动神经元的比例从体外培养6 - 14天时的约25%增加到体外培养第三周后超过80%。成熟神经元放电间隔比未成熟神经元短且更活跃;然而,放电频率的发育性增加并不显著。在体外培养6 - 14天的OTCs中,大多数自发活动神经元以规则方式缓慢放电。其余神经元以离散爆发形式发放动作电位,类似于体内发作间期活动。这些神经元的比例从体外培养6 - 14天时的约40%增加到体外培养第三周后超过80%。在体外发育过程中,平均爆发持续时间从3.5秒增加到约8秒,而平均爆发频率(0.7 - 1次/分钟)保持不变。活动剥夺的神经元放电频率低,以离散爆发形式发放动作电位,平均爆发频率为0.4次/分钟。自发活动神经元的比例、神经元放电的变异性以及外植体的活力要么不受活动阻断影响,要么在正常生长培养基中培养5 - 6天后恢复到对照值。我们得出结论,在大鼠新皮质的OTCs中,体外发育过程中SBA的缺失延迟了负责放电强度发育性增加的兴奋性机制的成熟。爆发式放电模式的发育受活动阻断的影响较小。