DOI QR코드

DOI QR Code

Mitochondria-Specific Monoclonal Antibodies in Eggs and Embryos of the Ascidian Halocynthia roretzi

  • Baek, Yong Han (Dept. of Marine Molecular Bioscience, Gangneung-Wonju National University) ;
  • Lee, Wang Jong (Dept. of Marine Molecular Bioscience, Gangneung-Wonju National University) ;
  • Kim, Gil Jung (Dept. of Marine Molecular Bioscience, Gangneung-Wonju National University)
  • Received : 2017.10.10
  • Accepted : 2017.11.20
  • Published : 2017.12.31

Abstract

Ascidian embryos have become an important model for embryological studies, offering a simple example for mechanisms of cytoplasmic components segregation. It is a well-known example that the asymmetric segregation of mitochondria into muscle lineage cells occurs during ascidian embryogenesis. However, it is still unclear which signaling pathway is involved in this process. To obtain molecular markers for studying mechanisms involved in the asymmetric distribution of mitochondria, we have produced monoclonal antibodies, Mito-1, Mito-2 and Mito-3, that specifically recognize mitochondria-rich cytoplasm in cells of the ascidian Halocynthia roretzi embryos. These antibodies stained cytoplasm like reticular structure in epidermis cells, except for nuclei, at the early tailbud stage. Similar pattern was observed in vital staining of mitochondria with DiOC2, a fluorescent probe of mitochondria. Immunostaining with these antibodies showed that mitochondria are evenly distributed in the animal hemisphere blastomeres at cleavage stages, whereas not in the vegetal hemisphere blastomeres. Mitochondria were transferred to the presumptive muscle and nerve cord lineage cells of the marginal zone in the vegetal hemisphere more than to the presumptive mesenchyme, notochord and endoderm lineage of the central zone. Therefore, it is suggested that these antibodies will be useful markers for studying mechanisms involved in the polarized distribution of mitochondria during ascidian embryogenesis.

Keywords

References

  1. Acton BM, Jurisicova A, Jurisica I, Casper RF (2004) Alterations in mitochondrial membrane potential during preimplantation stages of mouse and human embryo development. Mol Hum Reprod 10:23-32. https://doi.org/10.1093/molehr/gah004
  2. Alberts B, Bray D, Hopkin K, Johnson AD, Lewis J, Raff M, Roberts K, Walter P (2013) Essential Cell Biology. 4th ed. Garland Science, New York, pp 447-452.
  3. Coffman JA, Denegre JM (2007) Mitochondria, redox signaling and axis specification in metazoan embryos. Dev Biol 308:266-280. https://doi.org/10.1016/j.ydbio.2007.05.042
  4. Dumollard R, Duchen M, Carroll J (2007) The role of mitochondrial function in the oocyte and embryo. Curr Top Dev Biol 77:21-49.
  5. Egner A, Jakobs S, Hell SW (2002) Fast 100-nm resolution three-dimensional microscope reveals structural plasticity of mitochondria in live yeast. Proc Natl Acad Sci USA 99:3370-3375. https://doi.org/10.1073/pnas.052545099
  6. Friedman JR, Nunnari J (2014) Mitochondrial form and function. Nature 505:335-343. https://doi.org/10.1038/nature12985
  7. Fujiwara S, Satoh N (1990) Pattern of segregation of mitochondria into muscle lineage cells during embryogenesis of the ascidian Halocynthia roretzi. Dev Growth Differ 32:531-539. https://doi.org/10.1111/j.1440-169X.1990.00531.x
  8. Hirano T, Nishida H (1997) Developmental fates of larval tissues after metamorphosis in ascidian Halocynthia roretzi. I. Origin of mesodermal tissues of the juvenile. Dev Biol 192:199-210. https://doi.org/10.1006/dbio.1997.8772
  9. Hoitzing H, Johnston IG, Jones NS (2015) What is the function of mitochondrial networks? A theoretical assessment of hypotheses and proposal for future research. BioEssays 37:687-700. https://doi.org/10.1002/bies.201400188
  10. Kim GJ, Kumano G, Nishida H (2007) Cell fate polarization in ascidian mesenchyme/muscle precursors by directed FGF signaling and role for an additional ectodermal FGF antagonizing signal in notochord/nerve cord precursors. Development 134:1509-1518. https://doi.org/10.1242/dev.02825
  11. Kim GJ, Nishida H (1998) Monoclonal antibodies against differentiating mesenchyme cells in larvae of the ascidian Halocynthia roretzi. Zoolog Sci 15:553-559.
  12. Lawrence EJ, Boucher E, Mandato CA (2016) Mitochondria-cytoskeleton associations in mammalian cytokinesis. Cell Div 11:3. https://doi.org/10.1186/s13008-016-0015-4
  13. Nishida H (1987) Cell lineage analysis in ascidian embryos by intracellular injection of a tracer enzyme. III. Up to the tissue restricted stage. Dev Biol 121:526-541. https://doi.org/10.1016/0012-1606(87)90188-6
  14. Nishida H (1990) Determinative mechanisms in secondary muscle lineages of ascidian embryos: development of muscle-specific features in isolated muscle progenitor cells. Development 108:559-568.
  15. Roegiers F, Djediat C, Dumollard R, Rouviere C, Sardet C (1999) Phases of cytoplasmic and cortical reorganizations of the ascidian zygote between fertilization and first division. Development 126:3101-3117.
  16. Soltys BJ, Gupta RS (1992) Interrelationships of endoplasmic reticulum, mitochondria, intermediate filaments, and microtubules-a quadruple fluorescence labeling study. Biochem Cell Biol 70:1174-1186. https://doi.org/10.1139/o92-163
  17. Tang HL, Lung HL, Wu KC, Le AP, Tang HM, Fung MC (2007) Vimentin supports mitochondrial morphology and organization. Biochemical J 410:141-146.
  18. Van Blerkom J (2009) Mitochondria in early mammalian development. Semin Cell Dev Biol 20:354-364. https://doi.org/10.1016/j.semcdb.2008.12.005
  19. Zalokar M, Sardet C (1984) Tracing of cell lineage in embryonic development of Phallusia mammillata (Ascidia) by vital staining of mitochondria. Dev Biol 102:195-205. https://doi.org/10.1016/0012-1606(84)90184-2