During central nervous system (CNS) development neural stem cells (Neuroblasts, NBs) have to acquire an identity appropriate to their location. In thoracic and abdominal segments of Drosophila, the expression pattern of Bithorax-Complex Hox genes is known to specify the segmental identity of NBs prior to their delamination from the neuroectoderm. Compared to the thoracic, ground state segmental units in the head region are derived to different degrees, and the precise mechanism of segmental specification of NBs in this region is still unclear. We identified and characterized a set of serially homologous NB-lineages in the gnathal segments and used one of them (NB6-4 lineage) as a model to investigate the mechanism conferring segment-specific identities to gnathal NBs. We show that NB6-4 is primarily determined by the cell-autonomous function of the Hox gene Deformed ( Dfd). Interestingly, however, it also requires a non-cell-autonomous function of labial and Antennapedia that are expressed in adjacent anterior or posterior compartments. We identify the secreted molecule Amalgam (Ama) as a downstream target of the Antennapedia-Complex Hox genes labial, Dfd, Sex combs reduced and Antennapedia. In conjunction with its receptor Neurotactin (Nrt) and the effector kinase Abelson tyrosine kinase (Abl), Ama is necessary in parallel to the cell-autonomous Dfd pathway for the correct specification of the maxillary identity of NB6-4. Both pathways repress CyclinE ( CycE) and loss of function of either of these pathways leads to a partial transformation (40%), whereas simultaneous mutation of both pathways leads to a complete transformation (100%) of NB6-4 segmental identity. Finally, we provide genetic evidences, that the Ama-Nrt-Abl-pathway regulates CycE expression by altering the function of the Hippo effector Yorkie in embryonic NBs. The disclosure of a non-cell-autonomous influence of Hox genes on neural stem cells provides new insight into the process of segmental patterning in the developing CNS.
The central nervous system (CNS) needs to be subdivided into functionally specified regions. In the developing CNS of Drosophila, each neural stem cell, called neuroblasts (NB), acquires a unique identity according to its anterior-posterior and dorso-ventral position to generate a specific cell lineage. Along the anterior-posterior body axis, Hox genes of the Bithorax-Complex convey segmental identities to NBs in the trunk segments. In the derived gnathal and brain segments, the mechanisms specifying segmental NB identities are largely unknown. We investigated the role of Hox genes of the Antennapedia-Complex in the gnathal CNS. In addition to cell-autonomous Hox gene function, we unexpectedly uncovered a parallel non-cell-autonomous pathway in mediating segmental specification of embryonic NBs in gnathal segments. Both pathways restrict the expression of the cell cycle gene CyclinE, ensuring the proper specification of a glial cell lineage. Whereas the Hox gene Deformed mediates this cell-autonomously, labial and Antennapedia influence the identity via transcriptional regulation of the secreted molecule Amalgam (and its downstream pathway) in a non-cell-autonomous manner. These findings shed new light on the role of the highly conserved Hox genes during segmental patterning of neural stem cells in the CNS.