PMID:24466097

Citation	Liu, L and Wu, CF (2014) Distinct effects of abelson kinase mutations on myocytes and neurons in dissociated Drosophila embryonic cultures: mimicking of high temperature. PLoS ONE 9:e86438
Abstract	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.
Links	PubMed PMC3897706 Online version:10.1371/journal.pone.0086438
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
DROME:ABL		GO:0010592: positive regulation of lamellipodium assembly	ECO:0000315:		P		Compared to WT control, Abl neurons exhibited tendency to show fewer veil-like structures and well developed, large lamellipodia (>4 µm2) occurred only at a lower frequency (Fig. 5)	complete CACAO 9390
DROME:ABL		GO:0010977: negative regulation of neuron projection development	ECO:0000315:		P		Consistent with the altered neuronal growth in the larval CNS culture [24], neurons in Abl cultures displayed... more swollen, dark nodes along neurites (Fig. 4B1).	complete CACAO 9391
DROME:ABL	involved_in	GO:0010592: positive regulation of lamellipodium assembly	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
DROME:ABL	involved_in	GO:0010977: negative regulation of neuron projection development	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

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