Morphological and physiological properties of serotonergic neurons in dissociated cultures from the postnatal rat dorsal raphe nucleus.
Morphological and physiological properties of serotonergic neurons in dissociated cultures from the postnatal rat dorsal raphe nucleus.
复制标题
产后大鼠中缝背核分离培养物中血清素能神经元的形态和生理特性。
DOI：
10.1016/j.jneumeth.2007.08.018
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发表时间：
2008
影响因子：
3
通讯作者：
Nakajima,Yasuko
中科院分区：
文献类型：
--
作者：
Yasufuku-Takano,Junko;Nakajima,Shigehiro;Nakajima,Yasuko
我们已经开发了分离的原代培养中缝背核从出生后9-12天的大鼠。从脑切片中切出细胞核，分离，并在神经胶质饲养层上培养。5-羟色胺免疫细胞化学显示，62%的培养神经元是多巴胺能神经元。肾上腺素能神经元与非肾上腺素能神经元的直径无显著差异。用全细胞膜片钳方法检测培养神经元对5-HT 1A选择性激动剂8-羟二丙氨基四氢萘（8-OH-DPAT）的反应，然后用5-羟色胺免疫细胞化学方法处理。对8-OH-DPAT有反应的神经元中有92%是多巴胺能神经元。这些结果被用于鉴定多巴胺能神经元。在大多数情况下，多巴胺能神经元没有自发放电的动作电位。恒定电流去极化引起的动作电位列车，通常没有表现出显着的适应。应用8-OH-DPAT抑制动作电位放电。8-OH-DPAT诱导的电流的电流-电压关系表明其反向电位接近EK的内向整流。5-羟色胺能神经元被苯肾上腺素、蛙皮素和胃泌素释放肽去极化。该培养系统将作为一个有用的工具，阐明脑多巴胺能神经元的细胞，生理和分子特性。
We have developed dissociated primary cultures of the dorsal raphe nucleus from postnatal 9–12-day-old rats. The nucleus was dissected out from brain slices, dissociated, and cultured over a glial feeder layer. Serotonin immunocytochemistry revealed that 62% of cultured neurons were serotonergic. There was no significant difference in diameters between serotonergic and non-serotonergic neurons. With the whole-cell patch-clamp method, cultured neurons were tested for responses to 8-hydroxydipropylaminotetraline (8-OH-DPAT, a selective agonist for 5-HT1A), and then treated with serotonin immunocytochemistry. Ninety-two percent of neurons responding to 8-OH-DPAT were serotonergic. These results were used to identify serotonergic neurons. In most cases, serotonergic neurons did not show spontaneous firings of action potentials. Constant current depolarizations elicited trains of action potentials that usually did not show marked adaptation. Application of 8-OH-DPAT inhibited action potential firing. The current–voltage relation of the 8-OH-DPAT-induced current indicated an inward rectification with its reversal potential near EK. Serotonergic neurons were depolarized by phenylephrine, bombesin, and gastrin-releasing peptide. This culture system will serve as a useful tool for elucidating the cellular, physiological, and molecular properties of brain serotonergic neurons.