Single-cell Level Labelling Of Horizontal Motion Visual Neurons With Optimizing MARCM Condition

MARCM(Mosaic analysis with a repressible cell marker)is a genetic technique which is used to label single cell. It was invented by Tzumin Lee and Liqun Luo in 1999. MARCM allows one to positively label homozygous mutant cells through genetic recombination mediated by FRT. Up to now, MARCM has been applied in various fields of science: singlecell level labelling of neurons; lineage analysis of a single cell; generate homozygous mutations to investigate the function of gene in the healthy individuals. MARCM bring the level of the gene manipulation and neurology research to the single cell. MARCM also promote the rapid development of neuroscience. In technology, MARCM allows one to label cells only in one marker. On the basis of MARCM, Many chimeras techniques were invented. These techniques allows one to label different cells in double markers or multiple markers.We can get a neuroblast marked by GFP or a ganglion mother cell marked by GFP if active FLP when the neuroblast develop into the phase of mitosis. If the ganglion mother cell is not marked by GFP. Activate the recombinant enzyme FLP when it develop into the phase of mitosis. We can get two neurons and only one neuron is marked by GFP, so we can get a single cell clone. The development time of cell is very important for getting a single cell clone. In this paper, put virgin and male Drosophila in the vial with food. The Drosophila were moved to a new vial every two hours when the females begin laying eggs. Activate FLP at the development time of 50-70 hours in Drosophila eggs every two hours, and then count the amount of single-cell clone at the different time of activating FLP. We find the efficiency of labelling single cell of vertical system is the highest when active FLP after eggs of Drosophila have developed 60-62 h. However, the efficiency of labelling single cell of horizontal system is the highest when active FLP after eggs of Drosophila have developed 60-62 h.Use the MARCM technique to label single cell of horizontal system in the lobula plate of Drosophila. This article study the influence of sleep deprivation on the morphology of horizontal system neurons in the single-cell level. The study found that the dendritic length of horizontal system will increase significantly when the Drosophila have suffered from sleep deprivation for 12 h at night.