Brenneman D E, Neale E A, Habig W H, Bowers L M, Nelson P G
Brain Res. 1983 Jul;285(1):13-27. doi: 10.1016/0165-3806(83)90104-9.
Blockade of spontaneous electrical activity in dissociated fetal spinal cord cultures produced neuronal deficits as measured by biochemical and morphological techniques. Spinal cord cultures exhibited an age-dependent vulnerability to impulse blockade with tetrodotoxin (TTX) or xylocaine. Neuronal cell counts, [125I]tetanus toxin fixation and [125I]scorpion toxin binding indicated that TTX application produced neuronal deficits during the second or third week in culture. Application of TTX during the first or fourth week did not produce a difference in tetanus toxin fixation from controls. Radioautography of [125I]tetanus toxin revealed no obvious change in the label distribution after TTX treatment. Suppression of electrical activity during the first 6 days in culture had no effect on choline acetyltransferase (CAT) activity and no apparent effect on the appearance of the cultures. Application of TTX during the seventh day in culture decreased CAT activity to 68% of control. Chronic electrical blockade produced a progressively greater loss of CAT activity through 21 days in culture. GABAergic neurons, as indicated by high-affinity GABA uptake, glutamic acid decarboxylase activity and [3H]GABA radioautography, were not affected by electrical blockade. These data indicate that there is developmental and neurochemical specificity in the neuronal death produced by blocking spontaneous electrical activity in dissociated spinal cord cultures.
在解离的胎儿脊髓培养物中，通过生化和形态学技术测量发现，自发性电活动的阻断会导致神经元缺陷。脊髓培养物对河豚毒素（TTX）或利多卡因引起的冲动阻断表现出年龄依赖性的易损性。神经元细胞计数、[125I]破伤风毒素结合和[125I]蝎毒素结合表明，在培养的第二或第三周应用TTX会导致神经元缺陷。在第一周或第四周应用TTX，破伤风毒素结合与对照组相比没有差异。[125I]破伤风毒素的放射自显影显示，TTX处理后标记分布没有明显变化。在培养的前6天抑制电活动对胆碱乙酰转移酶（CAT）活性没有影响，对培养物的外观也没有明显影响。在培养的第七天应用TTX可使CAT活性降至对照组的68%。在培养21天的过程中，慢性电阻断导致CAT活性逐渐丧失。通过高亲和力GABA摄取、谷氨酸脱羧酶活性和[3H]GABA放射自显影表明，GABA能神经元不受电阻断的影响。这些数据表明，在解离的脊髓培养物中，阻断自发性电活动所导致的神经元死亡存在发育和神经化学特异性。
