Abstract
We have developed a simple and reliable method of preserving antigen immunoreactivity with concomitant excellent retention of the cell ultrastructure. Using this method, we have been able to follow the origin and developmental stages of nuage accumulations within the nurse cell/oocyte syncytium in the ovary of the fruit fly, Drosophila melanogaster, at the ultrastructural level. We have found two morphologically and biochemically distinct forms of nuage material in the nurse cell cytoplasm: translocating accumulations of nuage containing the Vasa protein, termed sponge bodies and stationary polymorphic accumulations of nuage enriched in Argonaute and Survival of motor neuron proteins. Immunogold labeling combined with confocal fluorescent and ultrastructural analyses have revealed that the Vasa-containing nuage accumulations remain closely associated with the cisternae of the endoplasmic reticulum throughout their lifetimes. The migration mechanism of the Vasa-positive nuage appears distinct from the microtubule-dependent translocation of oskar ribonucleoprotein complexes. We postulate that these two distinct nuage translocation pathways converge in the formation of the polar granules within the polar/germ plasm of the oocyte posterior pole. We also provide morphological and immunocytochemical evidence that these polymorphic nuage accumulations correspond to the recently described cytoplasmic domains termed U body-P body complexes.
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Acknowledgements
We are grateful to Prof. Janusz Kubrakiewicz (University of Wroclaw) for providing the confocal facilities and generously sharing ER-Tracker. We thank Dr. William W. Mattox (M.D. Anderson Cancer Center) for supplying Drosophila, Dr. Arnold Grabiec for help with the collection of the confocal data and Ms. Elzbieta Kisiel for preparing the figures. Finally, our thanks are due to anonymous reviewers for comments and suggestions that helped to improve this article.
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This research was supported by research grant K/ZDS/000791. M. Kloc is supported by NSF grant IOS-0904186.
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Jaglarz, M.K., Kloc, M., Jankowska, W. et al. Nuage morphogenesis becomes more complex: two translocation pathways and two forms of nuage coexist in Drosophila germline syncytia. Cell Tissue Res 344, 169–181 (2011). https://doi.org/10.1007/s00441-011-1145-2
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DOI: https://doi.org/10.1007/s00441-011-1145-2