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Abelson tyrosine kinase (Abl) is known to regulate axon guidance, muscle development, and cell-cell interaction  in vivo.  The  Drosophila  primary culture system offers advantages in exploring the cellular mechanisms mediated by Abl with utilizing various experimental manipulations. Here we demonstrate that single-embryo cultures exhibit stage-dependent characteristics of cellular differentiation and developmental progression in neurons and myocytes, as well as nerve-muscle contacts. In particular, muscle development critically depends on the stage of dissociated embryos. In wild-type (WT) cultures derived from embryos before stage 12, muscle cells remained within cell clusters and were rarely detected. Interestingly, abundant myocytes were spotted in  Abl  mutant cultures, exhibiting enhanced myocyte movement and fusion, as well as neuron-muscle contacts even in cultures dissociated from younger, stage 10 embryos. Notably,  Abl  myocytes frequently displayed well-expanded lamellipodia. Conversely,  Abl  neurons were characterized with fewer large veil-like lamellipodia, but instead had increased numbers of filopodia and darker nodes along neurites. These distinct phenotypes were equally evident in both homo- and hetero-zygous cultures ( Abl/Abl  vs.  Abl /+) of different alleles ( Abl 1  and Abl 4  ) indicating dominant mutational effects. Strikingly, in WT cultures derived from stage 10 embryos, high temperature (HT) incubation promoted muscle migration and fusion, partially mimicking the advanced muscle development typical of  Abl  cultures. However, HT enhanced neuronal growth with increased numbers of enlarged lamellipodia, distinct from the characteristic  Abl  neuronal morphology. Intriguingly, HT incubation also promoted  Abl  lamellipodia expansion, with a much greater effect on nerve cells than muscle. Our results suggest that Abl is an essential regulator for myocyte and neuron development and that high-temperature incubation partially mimics the faster muscle development typical of  Abl  cultures. Despite the extensive alterations by  Abl  mutations, we observed myocyte fusion events and nerve-muscle contact formation between WT and  Abl  cells in mixed WT and  Abl  cultures derived from labeled embryos.

Distinct Effects of Abelson Kinase Mutations on Myocytes and Neurons in Dissociated Drosophila Embryonic Cultures: Mimicking of High Temperature