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 an 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 an 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.
KeywordsDrosophila Embryonic brain Neural stem cell Neuroblast lineage DiI labeling Antibody staining
The authors thank Thomas Kunz, Martin Steimel, and Christof Rickert for sharing protocols and considerable expertise, and David Jussen for critically reading the manuscript. We are grateful to Gerd Technau for his general support. This work was supported by grants from the Deutsche Forschungsgemeinschaft (UR163/2-1 and UR163/3-1) and by a research stipend to K.F.K. from the FTN of the University Mainz.
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