Analysis of complete neuroblast cell lineages in the Drosophila embryonic brain via DiI labeling.
Proper functioning of the brain relies on an enormous diversity of neural cells generated by neural stem cell-like neuroblasts (NBs). Each of the about 100 NBs in each side of brain generates a nearly invariant and unique cell lineage, consisting of specific neural cell types that develop in defined time periods. In this chapter we describe a method that labels entire NB lineages in the embryonic brain. Clonal DiI labeling allows us to follow the development of a NB lineage starting from the neuroectodermal precursor cell up to the fully developed cell clone in the first larval instar brain. We also show how to ablate individual cells within a NB clone, which reveals information about the temporal succession in which daughter cells are generated. Finally, we describe how to combine clonal DiI labeling with fluorescent antibody staining that permits relating protein expression to individual cells within a labeled NB lineage. These protocols make it feasible to uncover precise lineage relationships between a brain NB and its daughter cells, and to assign gene expression to individual clonal cells. Such lineage-based information is a critical key for understanding the cellular and molecular mechanisms that underlie specification of cell fates in spatial and temporal dimension in the embryonic brain.
Institute of Genetics, University of Mainz, Mainz, Germany.
Karoline F
KF
Kraft
Rolf
R
Urbach
D000818:Animals; D001921:Brain / Q000166:cytology / Q000196:embryology*; D002232:Carbocyanines / Q000378:metabolism*; D019070:Cell Lineage; D002999:Clone Cells / Q000166:cytology / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology / Q000196:embryology*; D006536:Heptanes / Q000737:chemistry; D007814:Larva / Q000166:cytology; D054258:Neural Plate / Q000166:cytology; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D010063:Ovum / Q000166:cytology; D020959:Polyethylene / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors
C582073:1,1'-dioleyl-3,3,3',3'-tetramethylindocarbocyanine; D002232:Carbocyanines; D006536:Heptanes; D020959:Polyethylene