Localization of the Contacts Between Kenyon Cells and Aminergic Neurons in the Drosophila melanogaster Brain Using SplitGFP Reconstitution
Localization of the Contacts Between Kenyon Cells and Aminergic Neurons in the Drosophila melanogaster Brain Using SplitGFP Reconstitution
复制标题
DOI：
10.1002/cne.23388
复制
发表时间：
2013-12-01
影响因子：
2.5
通讯作者：
Fiala, Andre
中科院分区：
文献类型：
--
作者：
Pech, Ulrike;Pooryasin, Atefeh;Fiala, Andre
昆虫大脑的蘑菇体代表了一个参与控制适应行为的神经元回路，例如，联想学习其功能依赖于生物胺对凯尼恩细胞活性或突触递质释放的调节，例如，章鱼胺多巴胺或血清素因此，为了全面了解蘑菇体，不仅要确定哪些调节神经元与凯尼恩细胞相互作用，而且还要确定它们在蘑菇体中的确切位置。为了实现后者，我们利用GRASP技术，并创建了转基因果蝇黑腹果蝇，携带一部分的膜结合splitGFP在Kenyon细胞，沿着与胞质红色荧光标记。splitGFP的第二部分使用细胞特异性Gal 4驱动程序在不同的神经元群体中表达。只有当这些神经元与Kenyon细胞紧密接触时，GFP才能重建，这与双光子显微镜相结合，提供了非常高的空间分辨率。我们的特点空间和微观结构不同的接触区域之间的凯尼恩细胞和多巴胺，多巴胺能，章鱼胺能/酪胺能神经元的蘑菇体的所有细分。多巴胺能神经元的亚群密集地接触互补叶区域。章鱼胺能/酪胺能神经元稀疏地接触凯尼恩细胞，并且主要限于花萼、叶和叶。Kenyon细胞与多巴胺能神经元的接触不均匀地分布在整个蘑菇体上。总之，该技术使我们能够通过与胺能神经元的差分接触精确地定位蘑菇体的分段。J. Comp.神经元521：3992-4026，2013。(c)2013 Wiley Periodicals，Inc.
The mushroom body of the insect brain represents a neuronal circuit involved in the control of adaptive behavior, e.g., associative learning. Its function relies on the modulation of Kenyon cell activity or synaptic transmitter release by biogenic amines, e.g., octopamine, dopamine, or serotonin. Therefore, for a comprehensive understanding of the mushroom body, it is of interest not only to determine which modulatory neurons interact with Kenyon cells but also to pinpoint where exactly in the mushroom body they do so. To accomplish the latter, we made use of the GRASP technique and created transgenic Drosophila melanogaster that carry one part of a membrane-bound splitGFP in Kenyon cells, along with a cytosolic red fluorescent marker. The second part of the splitGFP is expressed in distinct neuronal populations using cell-specific Gal4 drivers. GFP is reconstituted only if these neurons interact with Kenyon cells in close proximity, which, in combination with two-photon microscopy, provides a very high spatial resolution. We characterize spatially and microstructurally distinct contact regions between Kenyon cells and dopaminergic, serotonergic, and octopaminergic/tyraminergic neurons in all subdivisions of the mushroom body. Subpopulations of dopaminergic neurons contact complementary lobe regions densely. Octopaminergic/tyraminergic neurons contact Kenyon cells sparsely and are restricted mainly to the calyx, the -lobes, and the -lobes. Contacts of Kenyon cells with serotonergic neurons are heterogeneously distributed over the entire mushroom body. In summary, the technique enables us to localize precisely a segmentation of the mushroom body by differential contacts with aminergic neurons. J. Comp. Neurol. 521:3992-4026, 2013. (c) 2013 Wiley Periodicals, Inc.