Ostrovsky Aaron, Cachero Sebastian, Jefferis Gregory
Cold Spring Harb Protoc. 2013 Apr 1;2013(4):335-41. doi: 10.1101/pdb.prot071712.
Clonal analysis with the MARCM (mosaic analysis with a repressible cell marker) system can be used for studying cell lineage, development, and anatomy in the Drosophila olfactory system and other parts of the fly brain. This protocol gives a method for generating flies with mosaic labeling. It describes how to establish a mating cage for MARCM in PNs (projection neurons) of the fly antennal lobe and then select appropriate flies for dissection and staining using immunohistochemistry. The protocol can be adapted to determine the birth order of neuroblast lineages or individual cells. Alternatively, it can be used to dissect a complicated Gal4 line into its component neuroblast lineages to help elucidate projection patterns and connectivity. Collecting newly hatched larvae during a short time window allows for precise control of the stage during development at which the heat shock is applied.
利用MARCM（可抑制细胞标记的镶嵌分析）系统进行的克隆分析可用于研究果蝇嗅觉系统及蝇脑其他部位的细胞谱系、发育和解剖结构。本方案提供了一种产生镶嵌标记果蝇的方法。它描述了如何在果蝇触角叶的投射神经元（PNs）中建立用于MARCM的交配笼，然后选择合适的果蝇进行解剖并用免疫组织化学染色。该方案可用于确定神经母细胞谱系或单个细胞的出生顺序。或者，它可用于将复杂的Gal4品系分解为其组成的神经母细胞谱系，以帮助阐明投射模式和连接性。在短时间窗口内收集刚孵化的幼虫，可精确控制施加热休克时的发育阶段。
