1] Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia, USA. [2] Kyorin University School of Medicine, Mitaka, Tokyo, Japan. [3].
1] Department of Neurobiology, University of Massachusetts, Worcester, Massachusetts, USA. [2] [3].
Nat Neurosci. 2014 Apr;17(4):631-7. doi: 10.1038/nn.3654. Epub 2014 Feb 23.
The Drosophila cerebrum originates from about 100 neuroblasts per hemisphere, with each neuroblast producing a characteristic set of neurons. Neurons from a neuroblast are often so diverse that many neuron types remain unexplored. We developed new genetic tools that target neuroblasts and their diverse descendants, increasing our ability to study fly brain structure and development. Common enhancer-based drivers label neurons on the basis of terminal identities rather than origins, which provides limited labeling in the heterogeneous neuronal lineages. We successfully converted conventional drivers that are temporarily expressed in neuroblasts, into drivers expressed in all subsequent neuroblast progeny. One technique involves immortalizing GAL4 expression in neuroblasts and their descendants. Another depends on loss of the GAL4 repressor, GAL80, from neuroblasts during early neurogenesis. Furthermore, we expanded the diversity of MARCM-based reagents and established another site-specific mitotic recombination system. Our transgenic tools can be combined to map individual neurons in specific lineages of various genotypes.
果蝇大脑起源于每个半球约 100 个神经母细胞，每个神经母细胞产生一组特征性的神经元。神经母细胞产生的神经元往往非常多样化，以至于许多神经元类型仍未被探索。我们开发了新的遗传工具，这些工具可以靶向神经母细胞及其多样化的后代，从而提高我们研究果蝇大脑结构和发育的能力。常见的基于增强子的驱动子根据神经元的终末身份而不是起源来标记神经元，这在异质神经元谱系中提供了有限的标记。我们成功地将传统的仅在神经母细胞中短暂表达的驱动子转化为在所有后续神经母细胞后代中表达的驱动子。一种技术涉及在神经母细胞及其后代中永生 GAL4 的表达。另一种方法依赖于在早期神经发生过程中从神经母细胞中丢失 GAL4 抑制剂 GAL80。此外，我们扩展了 MARCM 为基础的试剂的多样性，并建立了另一个特定的有丝分裂重组系统。我们的转基因工具可以组合使用，以在各种基因型的特定谱系中定位单个神经元。
