隔区、视前区和下丘脑区域神经元的无血清培养。三碘甲状腺原氨酸和雌二醇的作用。

A serum-free culture of the neurons in the septal, preoptic, and hypothalamic region. Effects of triiodothyronine and estradiol.

作者信息

Akuzawa K, Wakabayashi K

出版信息

Endocrinol Jpn. 1985 Feb;32(1):163-73. doi: 10.1507/endocrj1954.32.163.

DOI:10.1507/endocrj1954.32.163

Abstract

Serial modifications of Bottenstein and Sato' serum-free hormone-supplemented medium resulted in a new promising medium (1 : 1 mixture of L15 and MCDB 104 containing several supplements) for culturing neonatal rat brain cells. This medium favored the morphological and biochemical differentiation of the neurons, including particular types of cholinergic and cholinoceptive neurons, obtained enzymatically from the septum, preoptic area, and hypothalamus. On the other hand, the growth of non-neuronal cells was markedly suppressed in this medium. Therefore, their effects on the neurons are minimized in this culture. Effects of triiodothyronine (T3) and estradiol (E2) on the activities of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), synthetic and degradative enzymes for acetylcholine, respectively, were examined in this culture. The optimal concentrations of T3 and E2 for AChE activity were around 1 nM and 10 pM, respectively. However, E2 appeared to be somewhat inhibitory at higher concentrations. Although the activity of ChAT was maximum around 10 pM of E2, the ChAT activity increased as the concentration of T3 was increased to 100 nM.

摘要

对博滕施泰因和佐藤的无血清激素补充培养基进行系列改良后，得到了一种有前景的新培养基（L15和MCDB 104按1:1混合，并添加了几种补充成分），用于培养新生大鼠脑细胞。这种培养基有利于从隔区、视前区和下丘脑酶解获得的神经元的形态和生化分化，包括特定类型的胆碱能和胆碱感受神经元。另一方面，这种培养基能显著抑制非神经元细胞的生长。因此，在这种培养中它们对神经元的影响被最小化。在此培养中检测了三碘甲状腺原氨酸（T3）和雌二醇（E2）分别对胆碱乙酰转移酶（ChAT）和乙酰胆碱酯酶（AChE）活性的影响，ChAT和AChE分别是乙酰胆碱的合成酶和降解酶。AChE活性的T3和E2最佳浓度分别约为1 nM和10 pM。然而，较高浓度时E2似乎有一定抑制作用。虽然E2浓度约为10 pM时ChAT活性最高，但随着T3浓度增加到100 nM，ChAT活性也增加。