Wani Adil R, Hamid Aisha, Chaya Gonzalo N Morales, Syed Mubarak Hussain
Department of Biology, University of New Mexico, Albuquerque, New Mexico 87131, USA.
Department of Biology, University of New Mexico, Albuquerque, New Mexico 87131, USA
Cold Spring Harb Protoc. 2025 Apr 1;2025(4):pdb.prot108442. doi: 10.1101/pdb.prot108442.
From insects to humans, the nervous system generates complex behaviors mediated by distinct neural circuits that are composed of diverse cell types. During development, the spatiotemporal gene expression of the neural progenitors expands the diversity of neuronal and glial subtypes. Various neural stem cell-intrinsic and -extrinsic gene programs have been identified that are thought to play a major role in generating diverse neuronal and glial cell types. has served as an excellent model system for discovering the fundamental principles of nervous system development and function. The sophisticated genetic tools allow us to link the origin and birth timing (the time when a particular neuron is born during development) of neuron types to unique neural stem cells (NSCs) and to a developmental time. In , a special class of NSCs called Type II NSCs has adopted a more advanced division mode to generate lineages for the higher-order brain center, the central complex, which is an evolutionarily conserved brain region found in all insects. Type II NSCs, similar to the human outer radial glia, generate intermediate neural progenitors (INPs), which divide many times to produce about eight to 10 progeny. Both Type II NSCs and INPs express distinct transcription factors and RNA-binding proteins that have been proposed to regulate the specification of cell types populating the adult central complex. Here, we describe the recently invented lineage filtering system, called cell class-lineage intersection (CLIn), which enables the tracking and birthdating of the Type II NSC lineages. Using CLIn, one can easily generate clones of different Type II NSCs and identify not only the origins of neurons of interest but also their birth time.
从昆虫到人类，神经系统产生由不同神经回路介导的复杂行为，这些神经回路由多种细胞类型组成。在发育过程中，神经祖细胞的时空基因表达扩展了神经元和神经胶质细胞亚型的多样性。已经确定了各种神经干细胞内在和外在的基因程序，它们被认为在产生多样的神经元和神经胶质细胞类型中起主要作用。 已成为发现神经系统发育和功能基本原理的优秀模型系统。复杂的遗传工具使我们能够将神经元类型的起源和出生时间（特定神经元在发育过程中诞生的时间）与独特的神经干细胞（NSC）以及发育时间联系起来。在 中，一类特殊的神经干细胞，即II型神经干细胞，采用了更先进的分裂模式，为高阶脑中心——中央复合体生成谱系，中央复合体是在所有昆虫中都存在的进化保守脑区。II型神经干细胞类似于人类的外侧放射状胶质细胞，产生中间神经祖细胞（INP），后者多次分裂产生大约8到10个后代。II型神经干细胞和INP都表达独特的转录因子和RNA结合蛋白，这些蛋白被认为可以调节构成成年中央复合体的细胞类型的特化。在这里，我们描述了最近发明的谱系筛选系统，称为细胞类谱系交集（CLIn），它能够追踪II型神经干细胞谱系并确定其出生时间。使用CLIn，人们可以轻松地生成不同II型神经干细胞的克隆，不仅可以确定感兴趣神经元的起源，还可以确定它们的出生时间。
