Voltage-gated currents and firing properties of embryonic Drosophila neurons grown in a chemically defined medium.
Voltage-gated currents and firing properties of embryonic Drosophila neurons grown in a chemically defined medium.
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在化学成分确定的培养基中生长的胚胎果蝇神经元的电压门控电流和放电特性。
DOI：
10.1002/neu.480270111
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
O'Dowd,DK
中科院分区：
文献类型：
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作者：
O'Dowd,DK
本研究报道了一种化学成分确定的培养基（DDM 1）的组成，该培养基支持从原肠胚中期制备的分离细胞培养物中神经元的存活和分化。用神经特异性标记物染色的具有神经元形态的细胞在体外1天内明显分化，并且可以在培养物中维持长达2周。尽管在DDM 1中生长的神经元的全细胞电容测量值比在常规血清补充培养基中生长的神经元的全细胞电容测量值大5至10倍，但在两种生长条件下钾电流密度相似。与在补充血清的培养基中生长的神经元相比，在DDM 1中生长的神经元中观察到钠电流密度的小但显著的增加。大多数生长在DDM1中的神经元对去极化电流的注入产生单一或一连串的动作电位。在DDM 1中生长的单个神经元之间观察到的放电特性的异质性是电压依赖性钠、钙和钾电流的表达水平和门控特性的异质性。胚胎果蝇在化学成分确定的培养基中分化的能力以及它们适合电压钳和电流钳分析的事实使得该系统非常适合旨在了解参与电兴奋性的离子通道表达调控机制的研究。John Wiley & Sons，Inc.
This study reports the composition of a chemically defined medium (DDM1) that supports the survival and differentiation of neurons in dissociated cell cultures prepared from midgastrula stageDrosophilaembryos. Cells with neuronal morphology that stain with a neural‐specific marker are clearly differentiated by 1 dayin vitroand can be maintained in culture for up to 2 weeks. Although the whole cell capacitance measurements from neurons grown in DDM1 were 5‐ to 10‐fold larger than those of neurons grown in a conventional serum‐supplemented medium, the potassium current densities were similar in the two growth conditions. A small but significant increase in the sodium current density was observed in the neurons grown in DDM1 compared with those in serum‐supplemented medium. The majority of neurons grown in DDM1 fired either single or trains of action potentials in response to injection of depolarizing current. Contributing to the observed heterogeneity in the firing properties between individual neurons grown in DDM1 was heterogeneity in the levels of expression and gating properties of voltage‐dependent sodium, calcium, and pottassium currents. The ability of embryonicDrosophilaneurons to differentiate in a chemically defined medium and the fact that they are amenable to both voltage‐clamp and current‐clamp analysis makes this system well suited to studies aimed at understanding the mechanisms regulating expression of ion channels involved in electrical excitability. © 1995 John Wiley & Sons, Inc.