Brain reaggregate cultures in neurotoxicological investigations: adaptational and neuroregenerative processes following lesions.
Brain reaggregate cultures in neurotoxicological investigations: adaptational and neuroregenerative processes following lesions.
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神经毒理学研究中的大脑重新聚集培养物：病变后的适应和神经再生过程。
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发表时间：
1987
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通讯作者：
A. Ck
中科院分区：
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作者：
A. Ck
体外神经系统可以预测中枢神经系统毒性，除非外源药物主要影响血脑屏障。神经生物学研究中广泛使用的系统可用于机械神经毒理学研究，尽管系统的选择通常是任意的。一种更理性的方法现在可能是合理的。有许多可用的培养系统，包括神经细胞系、器官型外植体或再聚集培养，以及单个神经细胞类型的原代单层培养：神经元、星形胶质细胞和少突胶质细胞。在这些模型中，最近使用器官型外植体培养型取得了很大的成功。同样，在我们的实验室中，使用大鼠全脑再聚集体培养，我们已经证明了胆碱能神经毒性乙胆碱芥末aziridinium （ECMA）在体外/体内具有良好的相关性，其中使用低浓度的ECMA（12.5微米）产生特定的胆碱能病变。如对小脑谷氨酸能颗粒神经元的非特异性作用所示，高浓度（25-50微米）具有更强的细胞毒性。用神经营养因子，神经生长因子（NGF）治疗胆碱毒素损伤的重聚集体，并没有逆转病变，但用NGF （50 ng/ml）治疗对照细胞，胆碱乙酰转移酶（ChAT）活性和毒蕈碱受体结合均升高。因此，“受损”的再聚集培养系统可能在评估潜在的治疗药物方面具有未来的价值，这些药物可以逆转中枢神经系统的这种病变。通过利用神经元或星形胶质细胞的原代单层培养来补充在重组系统中获得的信息，我们可以提出一种体外潜在神经毒物的逐步筛选系统。其最简单的形式是：(1)使用肿瘤来源的神经细胞系进行初步筛选，(2)在全脑再聚集体中测试选定的化合物，(3)用单个神经细胞类型的初级单层培养补充信息。
In vitro neural systems can be predictive for CNS neurotoxicity, except where xenobiotics primarily affect the blood-brain barrier. The wide range of systems now used in neurobiological studies is available for mechanistic neurotoxicological investigations although the choice of system is generally arbitrary. A more rational approach may now be justified. There are many culture systems available including neural cell lines, organotypic explant or reaggregation cultures, and primary monolayer cultures of individual neural cell types: neurons, astrocytes, and oligodendrocytes. Of these models much success has recently been achieved using the organotypic explant culture type. Similarly in our laboratories, using rat whole-brain reaggregate cultures, we have demonstrated good in vitro/in vivo correlations for the cholinergic neurotoxicant ethylcholine mustard aziridinium (ECMA) where specific cholinergic lesions are produced using low concentrations of ECMA (12.5 microM). Higher concentrations (25-50 microM) were more cytotoxic, as shown, for example, by nonspecific effects on cerebellar glutamatergic granule neurons. Treatment of reaggregates lesioned with the cholinotoxin with a neurotrophic factor, nerve growth factor (NGF), did not reverse the lesion but treatment of control cells with NGF (50 ng/ml) elevated both choline acetyltransferase (ChAT) activity and muscarinic receptor binding. The "lesioned" reaggregate culture system may thus be of future value in evaluating potential therapeutic agents that could reverse such lesions in the CNS. By supplementing the information gained in the reaggregate system with tests using primary monolayer cultures of neurons or astrocytes we can propose a stepwise screening system for potential neurotoxicants in vitro. In its simplest form this is (1) screen initially using tumor-derived neural cell line, (2) test selected compounds in whole-brain reaggregates, and (3) supplement information with primary monolayer cultures of individual neural cell types.