The embryonic central nervous system lineages of Drosophila melanogaster. II. Neuroblast lineages derived from the dorsal part of the neuroectoderm.

In Drosophila, central nervous system (CNS) formation starts with the delamination from the neuroectoderm of about 30 neuroblasts (NBs) per hemisegment. They give rise to approximately 350 neurons and 30 glial cells during embryonic development. Understanding the mechanisms leading to cell fate specification and differentiation in the CNS requires the identification of the NB lineages. The embryonic lineages derived from 17 NBs of the ventral part of the neuroectoderm have previously been described (Bossing et al., 1996). Here we present 13 lineages derived from the dorsal part of the neuroectoderm and we assign 12 of them to identified NBs. Together, the 13 lineages comprise approximately 120 neurons and 22 to 27 glial cells which we include in a systematic terminology. Therefore, NBs from the dorsal neuroectoderm produce about 90% of the glial cells in the embryonic ventral ganglion. Two of the NBs give rise to glial progeny exclusively (NB 6-4A, GP) and five to glia as well as neurons (NBs 1-3, 2-5, 5-6, 6-4T, 7-4). These seven NBs are arranged as a group in the most lateral region of the NB layer. The other lineages (NBs 2-4, 3-3, 3-5, 4-3, 4-4, 5-4, clone y) are composed exclusively of neurons (interneurons, motoneurons, or both). Additionally, it has been possible to link the lateral cluster of even-skipped expressing cells (EL) to the lineage of NB 3-3. Along with the previously described clones, the vast majority (more than 90%) of cell lineages in the embryonic ventral nerve cord (thorax, abdomen) are now known. Moreover, previously identified neurons and most glial cells are now linked to certain lineages and, thus, to particular NBs. This complete set of data provides a foundation for the interpretation of mutant phenotypes and for future investigations on cell fate specification and differentiation.

The differentiation of the serotonergic neurons in the Drosophila ventral nerve cord depends on the combined function of the zinc finger proteins Eagle and Huckebein.

The Drosophila ventral nerve cord (vNC) derives from a stereotyped population of neural stem cells, neuroblasts (NBs), each of which gives rise to a characteristic cell lineage. The mechanisms leading to the specification and differentiation of these lineages are largely unknown. Here we analyse mechanisms leading to cell differentiation within the NB 7-3 lineage. Analogous to the grasshopper, NB 7-3 is the progenitor of the Drosophila vNC serotonergic neurons. The zinc finger protein Eagle (Eg) is expressed in NB 7-3 just after delamination and is present in all NB 7-3 progeny until late stage 17. DiI cell lineage tracing and immunocytochemistry reveal that eg is required for normal pathfinding of interneuronal projections and for restricting the cell number in the thoracic NB 7-3 lineage. Moreover, eg is required for serotonin expression. Ectopic expression of Eg protein forces specific additional CNS cells to enter the serotonergic differentiation pathway. Like NB 7-3, the progenitor(s) of these ectopic cells express Huckebein (Hkb), another zinc finger protein. However, their progenitors do not express engrailed (en) as opposed to the NB 7-3 lineage, where en acts upstream of eg. We conclude that eg and hkb act in concert to determine serotonergic cell fate, while en is more distantly involved in this process by activating eg expression. Thus, we provide the first functional evidence for a combinatorial code of transcription factors acting early but downstream of segment polarity genes to specify a unique neuronal cell fate.

The origin, location, and projections of the embryonic abdominal motorneurons of Drosophila.

We have used a retrograde labeling technique to identify motorneurons for each of the 30 body wall muscles of an abdominal hemisegment in the late stage 16 Drosophila embryo. Each motorneuron has a characteristic cell body position, dendritic arborization, and axonal projection. In addition, we have determined the neuroblasts of origin for most of the motorneurons we describe. Some organizational principles for the neuromuscular system have become apparent: (1) There is no obvious topographic relationship between the cell body positions of motorneurons and the position or orientation of the muscles they innervate; (2) motorneurons that innervate muscles of similar position and orientation are often clustered and have overlapping dendritic trees; (3) morphologically similar motorneurons are generally derived from a common neuroblast and innervate operationally related muscles; and (4) neuroblasts can give rise to more than one morphological type of motorneuron.

The embryonic central nervous system lineages of Drosophila melanogaster. I. Neuroblast lineages derived from the ventral half of the neuroectoderm.

Central nervous system development in Drosophila starts with the delamination from the neuroectoderm of about 30 neuroblasts (NBs) per hemisegment. Understanding the mechanisms leading to the specification of the individual NBs and their progeny requires the identification of their lineages. Here we describe 17 embryonic NB lineages derived from the ventral half of the neuroectoderm and we assign these lineages to identified medial and intermediate NBs. The lineages are composed of interneurons (NB 1-2, NB 2-1, MP2, NB 4-1, NB 5-1, NB 5-3, NB 6-1, NB 6-2, and NB 7-2), interneurons and motoneurons (NB 3-1, NB 3-2, NB 4-2, NB 5-2, NB 7-1, and NB 7-3), or interneurons, motoneurons, and glial cells (NB 1-1 and NB 2-2). NB 1-1, NB 2-2, and NB 3-1 form segment-specific lineages. Neuroectodermal progenitors forming NB 2-1, NB 5-1, and NB 7-3 divide while still in the ectoderm to give rise to an additional epidermoblast. Expression of segmentation genes is not lineal in the clones of NB 1-2 and NB 7-3 (engrailed), NB 1-1, NB 4-2, and NB 7-1 (even-skipped), and NB 7-1 (gooseberry-proximal). The timing of delamination for individual NBs as well as the number of their progeny is not strictly invariant. The 17 NBs produce about 200 neurons and only three glial cells, corresponding to about 70% of the estimated total number of neurons and 10% of the glial cells per thoracic and abdominal hemisegment. Previously identified neural cell types were linked to particular lineages and we introduce a systematic terminology for the ventral nerve cord neurons. The wild-type clones provide a foundation for the analysis of mutants, expression patterns, and experimental manipulations.

huckebein is required for glial development and axon pathfinding in the neuroblast 1-1 and neuroblast 2-2 lineages in the Drosophila central nervous system.

huckebein encodes a predicted zinc finger transcription factor which is transiently expressed in a subset of Drosophila central nervous system precursors (neuroblasts (NBs)). We used DiI cell lineage tracing and cell fate markers to investigate the role of huckebein in the NB 1-1 and NB 2-2 cell lineages. Loss of huckebein does not switch these NBs into different NB fates, nor does it change the number of cells in their lineages; rather, it is required for glial development in the NB 1-1 lineage, and for axon pathfinding of a subset of interneurons and motoneurons in both lineages.

New neuroblast markers and the origin of the aCC/pCC neurons in the Drosophila central nervous system.

Drosophila is an ideal system for identifying genes that control central nervous system (CNS) development. Particularly useful tools include molecular markers for subsets of neural precursors (neuroblasts) and the simple expression pattern of the even-skipped (eve) gene in a subset of neurons. Here we provide additional molecular markers for identified neuroblasts, including several with near single cell specificity. In addition, we use these new markers to trace the development of several eve+ neurons. Our results shows that the eve+ aCC/pCC neurons develop from a different neuroblast than previously thought, and have led us to assign new names for several neuroblasts. These results are supported by DiI cell lineage analysis of neuroblasts identified in vivo.

Commitment of CNS progenitors along the dorsoventral axis of Drosophila neuroectoderm.

In the Drosophila embryo, the central nervous system (CNS) develops from a population of neural stem cells (neuroblasts) and midline progenitor cells. Here, the fate and extent of determination of CNS progenitors along the dorsoventral axis was assayed. Dorsal neuroectodermal cells transplanted into the ventral neuroectoderm or into the midline produced CNS lineages consistent with their new position. However, ventral neuroectodermal cells and midline cells transplanted to dorsal sites of the neuroectoderm migrated ventrally and produced CNS lineages consistent with their origin. Thus, inductive signals at the ventral midline and adjacent neuroectoderm may confer ventral identities to CNS progenitors as well as the ability to assume and maintain characteristic positions in the developing CNS. Furthermore, ectopic transplantations of wild-type midline cells into single minded (sim) mutant embryos suggest that the ventral midline is required for correct positioning of the cells.

The fate of the CNS midline progenitors in Drosophila as revealed by a new method for single cell labelling.

We present a new method for marking single cells and tracing their development through embryogenesis. Cells are labelled with a lipophilic fluorescent tracer (DiI) in their normal positions without impaling their membranes. The dye does not diffuse between cells but is transferred to the progeny, disclosing their morphology in all detail. Behaviour of labelled cells can be observed in vivo (cell divisions, morphogenetic movements and differentiation). Following photoconversion of the dye, fully differentiated clones can be analyzed in permanent preparations. We apply this method for cell lineage analysis of the embryonic Drosophila CNS. Here we describe the fate of the CNS midline cells. We present the complete lineages of these cells in the fully differentiated embryo and show that variability exists in segmental numbers of the midline progenitors as well as in the composition of their lineages.

A common precursor for glia and neurons in the embryonic CNS of Drosophila gives rise to segment-specific lineage variants.

The nervous system consists of two classes of cells, neurons and glia, which differ in morphology and function. They derive from precursors located in the neurogenic region of the ectoderm. In this study, we present the complete embryonic lineage of a neuroectodermal precursor in Drosophila that gives rise to neurons as well as glia in the abdominal CNS. This lineage is conserved among different Drosophila species. We show that neuronal and glial cell types in this clone derive from one segregating precursor, previously described as NB1-1. Thus, in addition to neuroblasts and glioblasts, there exists a third class of CNS precursors in Drosophila, which we call neuroglioblasts. We further show that the NB 1-1 lineage exhibits characteristic segment-specific differences on the cellular level.