Time-lapse and cell ablation reveal the role of cell interactions in fly glia migration and proliferation

Institut de Génétique et Biologie Moléculaire et Cellulaire, IGBMC/CNRS/ULP/INSERM - BP 10142, ILLKIRCH, C. U. de Strasbourg 67404, France.
Development (Impact Factor: 6.46). 11/2004; 131(20):5127-38. DOI: 10.1242/dev.01398
Source: PubMed


Migration and proliferation have been mostly explored in culture systems or fixed preparations. We present a simple genetic model, the chains of glia moving along fly wing nerves, to follow such dynamic processes by time-lapse in the whole animal. We show that glia undergo extensive cytoskeleton and mitotic apparatus rearrangements during division and migration. Single cell labelling identifies different glia: pioneers with high filopodial, exploratory, activity and, less active followers. In combination with time-lapse, altering this cellular environment by genetic means or cell ablation has allowed to us define the role of specific cell-cell interactions. First, neurone-glia interactions are not necessary for glia motility but do affect the direction of migration. Second, repulsive interactions between glia control the extent of movement. Finally, autonomous cues control proliferation.

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    • "Ablation experiments were performed to confirm that additional glial nuclei along the NER of larvae originate from ePG2. High-energy UV irradiation was used to eliminate ePG2 in the embryo, a method previously used to ablate glial cells in the developing fly wing (Aigouy et al., 2008; Aigouy et al., 2004). "
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    ABSTRACT: One of the numerous functions of glial cells in Drosophila is the ensheathment of neurons to isolate them from the potassium-rich haemolymph, thereby establishing the blood-brain barrier. Peripheral nerves of flies are surrounded by three distinct glial cell types. Although all embryonic peripheral glia (ePG) have been identified on a single-cell level, their contribution to the three glial sheaths is not known. We used the Flybow system to label and identify each individual ePG in the living embryo and followed them into third instar larva. We demonstrate that all ePG persist until the end of larval development and some even to adulthood. We uncover the origin of all three glial sheaths and describe the larval differentiation of each peripheral glial cell in detail. Interestingly, just one ePG (ePG2) exhibits mitotic activity during larval stages, giving rise to up to 30 glial cells along a single peripheral nerve tract forming the outermost perineurial layer. The unique mitotic ability of ePG2 and the layer affiliation of additional cells were confirmed by in vivo ablation experiments and layer-specific block of cell cycle progression. The number of cells generated by this glial progenitor and hence the control of perineurial hyperplasia correlate with the length of the abdominal nerves. By contrast, the wrapping and subperineurial glia layers show enormous hypertrophy in response to larval growth. This characterisation of the embryonic origin and development of each glial sheath will facilitate functional studies, as they can now be addressed distinctively and genetically manipulated in the embryo.
    Development 07/2013; 140(17). DOI:10.1242/dev.093245 · 6.46 Impact Factor
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    • "Collective or chain migration is a fundamental feature of migration in Drosophila wing and peripheral nerve (Giangrande, 1994; Sepp et al., 2000; Aigouy et al., 2004; von Hilchen et al., 2008), in the Manduca enteric nervous system (Copenhaver and Taghert, 1989; Wright and Copenhaver, 2000) and in zebrafish peripheral nerves (Kucenas et al., 2008). In the moth antennal nerve, AN glial cells often appear in chains in vivo especially at the early stages of migration although the dimensions of the nerve and the pattern of glial process extension indicate that chaining along cannot be responsible for continuous ensheathment of axon bundles. "
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    ABSTRACT: In adult olfactory nerves of mammals and moths, a network of glial cells ensheathes small bundles of olfactory receptor axons. In the developing antennal nerve (AN) of the moth Manduca sexta, the axons of olfactory receptor neurons (ORNs) migrate from the olfactory sensory epithelium toward the antennal lobe. Here we explore developmental interactions between ORN axons and AN glial cells. During early stages in AN glial-cell migration, glial cells are highly dye coupled, dividing glia are readily found in the nerve and AN glial cells label strongly for glutamine synthetase. By the end of this period, dye-coupling is rare, glial proliferation has ceased, glutamine synthetase labeling is absent, and glial processes have begun to extend to enwrap bundles of axons, a process that continues throughout the remainder of metamorphic development. Whole-cell and perforated-patch recordings in vivo from AN glia at different stages of network formation revealed two potassium currents and an R-like calcium current. Chronic in vivo exposure to the R-type channel blocker SNX-482 halted or greatly reduced AN glial migration. Chronically blocking spontaneous Na-dependent activity by injection of tetrodotoxin reduced the glial calcium current implicating an activity-dependent interaction between ORNs and glial cells in the development of glial calcium currents.
    Neuron Glia Biology 09/2011; 6(4):245-61. DOI:10.1017/S1740925X11000081 · 6.64 Impact Factor
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    • "Previous studies on other insects have shown that apoptosis sculpts the peripheral region of the pupal wings in Lepidoptera including butterflies and moths (Dohrmann and Nihjout, 1988; Kodama et al., 1995) and that ecdysone signaling is involved in this process (Fujiwara and Ogai, 2001). In the case of Drosophila, although the apoptosis along the pupal wing margin has been described as in other insects, its purpose and underlying mechanism were not well understood (Aigouy et al., 2004; Jafar-Nejad et al., 2006). In this study, we showed that two distinct surges of apoptosis occur in Drosophila pupal wing development. "
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    ABSTRACT: Animal tissues and organs are comprised of several types of cells, which are often arranged in a well-ordered pattern. The posterior part of the Drosophila wing margin is covered with a double row of long hairs, which are equally and alternately derived from the dorsal and ventral sides of the wing, exhibiting a zigzag pattern in the lateral view. How this geometrically regular pattern is formed has not been fully understood. In this study, we show that this zigzag pattern is created by rearrangement of wing margin cells along the dorsoventral boundary flanked by the double row of hair cells during metamorphosis. This cell rearrangement is induced by selective apoptosis of wing margin cells that are spatially separated from hair cells. As a result of apoptosis, the remaining wing margin cells are rearranged in a well-ordered manner, which shapes corrugated lateral sides of both dorsal and ventral edges to interlock them for zigzag patterning. We further show that the corrugated topology of the wing edges is achieved by cell-type specific expression and localization of four kinds of NEPH1/nephrin family proteins through heterophilic adhesion between wing margin cells and hair cells. Homophilic E-cadherin adhesion is also required for attachment of the corrugated dorsoventral edges. Taken together, our results demonstrate that sequential coordination of apoptosis and epithelial architecture with selective adhesion creates the zigzag hair alignment. This may be a common mechanism for geometrically ordered repetitive packing of several types of cells in similarly patterned developmental fields such as the mammalian organ of Corti.
    Developmental Biology 07/2011; 357(2):336-46. DOI:10.1016/j.ydbio.2011.07.007 · 3.55 Impact Factor
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