Peter Grünberg Institute, Institute of Complex Systems, Bioelectronics, Forschungszentrum Jülich, 52425, Jülich, Germany.
J Mol Histol. 2012 Aug;43(4):405-19. doi: 10.1007/s10735-012-9395-1. Epub 2012 Mar 9.
We have designed a cell culture system for thoracic neurons of adult Locusta migratoria that enables the establishment of functional synapses in vitro. Patch-clamp recordings revealed three different neuron classes. About half of the neurons (47%) had unexcitable somata with outward and no inward conductance. The other half generated either single (37%) or multiple action potentials (18%) and differed mainly in lower outward conductance. Selectively stained motor neurons were analyzed to demonstrate varied physiological properties due to culture conditions. Using paired patch clamp recordings we demonstrate directly synaptic transmission in morphologically connected neurons in vitro. Presynaptic stimulation resulted in postsynaptic potentials in 42 pairs of neurons tested, independent of the type of neuron. According to pharmacological experiments most of these synapses were either glutamatergic or GABAergic. In addition to these chemical synapses, electrical synapses were found. With the demonstration of synapse formation in cell culture of adult locust neurons, this study provides the basis for the future analysis of more defined insect neuronal circuits in culture.
我们设计了一种用于成年蝗虫胸神经元的细胞培养系统，使体外建立功能性突触成为可能。膜片钳记录显示了三种不同的神经元类型。大约一半的神经元（47%）具有无兴奋性胞体，表现为外向而无内向电导。另一半神经元产生单一（37%）或多个动作电位（18%），主要区别在于较低的外向电导。选择性染色的运动神经元被分析以证明由于培养条件导致的不同生理特性。使用配对膜片钳记录，我们直接证明了体外形态连接神经元之间的突触传递。在测试的 42 对神经元中，无论神经元类型如何，突触前刺激均导致突触后电位。根据药理学实验，这些突触大部分是谷氨酸能或 GABA 能的。除了这些化学突触外，还发现了电突触。通过证明成年蝗虫神经元细胞培养中的突触形成，本研究为未来在培养中分析更明确的昆虫神经元回路提供了基础。
