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Pregnenolone stabilizes microtubules and promotes zebrafish embryonic cell movement

Nature volume 439, pages 480–483 (2006)Cite this article

Abstract

Embryonic cell movement is essential for morphogenesis and the establishment of body shapes1,2, but little is known about its mechanism. Here we report that pregnenolone, which is produced from cholesterol by the steroidogenic enzyme Cyp11a1 (cholesterol side-chain cleavage enzyme, P450scc)3, functions in promoting cell migration during epiboly. Epiboly is a process in which embryonic cells spread from the animal pole to cover the underlying yolk. During epiboly, cyp11a1 is expressed in an extra-embryonic yolk syncytial layer4. Reducing cyp11a1 expression in zebrafish using antisense morpholino oligonucleotides did not perturb cell fates, but caused epibolic delay. This epibolic defect was partially rescued by the injection of cyp11a1 RNA or the supplementation of pregnenolone. We show that the epibolic delay is accompanied by a decrease in the level of polymerized microtubules, and that pregnenolone can rescue this microtubule defect. Our results indicate that pregnenolone preserves microtubule abundance and promotes cell movement during epiboly.

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Figure 1: Two steroidogenic genes, cyp11a1 and hsd3b , are expressed in blastomeres and the yolk syncytial layer during early zebrafish embryogenesis.
Figure 2: Reduced cyp11a1 expression results in epibolic delay.
Figure 3: Pregnenolone rescues epiboly defect and increases microtubule content.
Figure 4: Fluorescein-conjugated pregnenolone co-localizes with microtubules.

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References

  1. Locascio, A. & Nieto, M. A. Cell movements during vertebrate development: integrated tissue behaviour versus individual cell migration. Curr. Opin. Genet. Dev. 11, 464–469 (2001)

    Article  CAS  Google Scholar 

  2. Myers, D. C., Sepich, D. S. & Solnica-Krezel, L. Convergence and extension in vertebrate gastrulae: cell movements according to or in search of identity? Trends Genet. 18, 447–455 (2002)

    Article  CAS  Google Scholar 

  3. Miller, W. L. Molecular biology of steroid hormone synthesis. Endocr. Rev. 9, 295–318 (1988)

    Article  CAS  Google Scholar 

  4. Hsu, H. J., Hsiao, P., Kuo, M. W. & Chung, B. C. Expression of zebrafish cyp11a1 as a maternal transcript and in yolk syncytial layer. Gene Expr. Patterns 2, 219–222 (2002)

    Article  CAS  Google Scholar 

  5. Chen, S. & Kimelman, D. The role of the yolk syncytial layer in germ layer patterning in zebrafish. Development 127, 4681–4689 (2000)

    CAS  PubMed  Google Scholar 

  6. Babin, P. J. et al. Both apolipoprotein E and A-I genes are present in a nonmammalian vertebrate and are highly expressed during embryonic development. Proc. Natl. Acad. Sci. USA 94, 8622–8627 (1997)

    Article  ADS  CAS  Google Scholar 

  7. Sharma, M. K. et al. Sequence, linkage mapping and early developmental expression of the intestinal-type fatty acid-binding protein gene (fabp2) from zebrafish (Danio rerio). Comp. Biochem. Physiol. B Biochem. Mol. Biol. 138, 391–398 (2004)

    Article  Google Scholar 

  8. Grigoryev, D. N. et al. Cytochrome P450c17-expressing Escherichia coli as a first-step screening system for 17α-hydroxylase-C17,20-lyase inhibitors. Anal. Biochem. 267, 319 (1999)

    Article  CAS  Google Scholar 

  9. Rheaume, E. et al. Structure and expression of a new complementary DNA encoding the almost exclusive 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase in human adrenals and gonads. Mol. Endocrinol. 5, 1147–1157 (1991)

    Article  CAS  Google Scholar 

  10. Jesuthasan, S. & Stahle, U. Dynamic microtubules and specification of the zebrafish embryonic axis. Curr. Biol. 7, 31–42 (1997)

    Article  CAS  Google Scholar 

  11. Solnica-Krezel, L. & Driever, W. Microtubule arrays of the zebrafish yolk cell: organization and function during epiboly. Development 120, 2443–2455 (1994)

    CAS  PubMed  Google Scholar 

  12. Strahle, U. & Jesuthasan, S. Ultraviolet irradiation impairs epiboly in zebrafish embryos: evidence for a microtubule-dependent mechanism of epiboly. Development 119, 909–919 (1993)

    CAS  PubMed  Google Scholar 

  13. Akhmanova, A. et al. Clasps are CLIP-115 and -170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts. Cell 104, 923–935 (2001)

    Article  CAS  Google Scholar 

  14. Liu, L., Tommasi, S., Lee, D. H., Dammann, R. & Pfeifer, G. P. Control of microtubule stability by the RASSF1A tumour suppressor. Oncogene 22, 8125–8136 (2003)

    Article  CAS  Google Scholar 

  15. Dent, E. W. & Gertler, F. B. Cytoskeletal dynamics and transport in growth cone motility and axon guidance. Neuron 40, 209–227 (2003)

    Article  CAS  Google Scholar 

  16. Watanabe, T., Noritake, J. & Kaibuchi, K. Regulation of microtubules in cell migration. Trends Cell Biol. 15, 76–83 (2005)

    Article  CAS  Google Scholar 

  17. Morris, N. R. Nuclear positioning: the means is at the ends. Curr. Opin. Cell Biol. 15, 54–59 (2003)

    Article  CAS  Google Scholar 

  18. Wittmann, T., Hyman, A. & Desai, A. The spindle: a dynamic assembly of microtubules and motors. Nature Cell Biol. 3, E28–E34 (2001)

    Article  CAS  Google Scholar 

  19. Murakami, K., Fellous, A., Baulieu, E. E. & Robel, P. Pregnenolone binds to microtubule-associated protein 2 and stimulates microtubule assembly. Proc. Natl Acad. Sci. USA 97, 3579–3584 (2000)

    Article  ADS  CAS  Google Scholar 

  20. D'Amico, L. A. & Cooper, M. S. Morphogenetic domains in the yolk syncytial layer of axiating zebrafish embryos. Dev. Dyn. 222, 611–624 (2001)

    Article  CAS  Google Scholar 

  21. Arensburg, J., Payne, A. H. & Orly, J. Expression of steroidogenic genes in maternal and extraembryonic cells during early pregnancy in mice. Endocrinology 140, 5220–5232 (1999)

    Article  CAS  Google Scholar 

  22. Hu, M. C. et al. Steroid deficiency syndromes in mice with targeted disruption of Cyp11a1. Mol. Endocrinol. 16, 1943–1950 (2002)

    Article  CAS  Google Scholar 

  23. Venihaki, M., Carrigan, A., Dikkes, P. & Majzoub, J. A. Circadian rise in maternal glucocorticoid prevents pulmonary dysplasia in fetal mice with adrenal insufficiency. Proc. Natl. Acad. Sci. USA 97, 7336–7341 (2000)

    Article  ADS  CAS  Google Scholar 

  24. Kozlova, T. & Thummel, C. S. Essential roles for ecdysone signaling during Drosophila mid-embryonic development. Science 301, 1911–1914 (2003)

    Article  ADS  CAS  Google Scholar 

  25. Cato, A. C., Nestl, A. & Mink, S. Rapid actions of steroid receptors in cellular signaling pathways. Sci. STKE 2002, RE9 (2002)

    PubMed  Google Scholar 

  26. Schmidt, B. M. et al. Rapid, nongenomic steroid actions: A new age? Front. Neuroendocrinol. 21, 57–94 (2000)

    Article  CAS  Google Scholar 

  27. Chiang, E. F. et al. Two sox9 genes on duplicated zebrafish chromosomes: expression of similar transcription activators in distinct sites. Dev. Biol. 231, 149–163 (2001)

    Article  CAS  Google Scholar 

  28. Gard, D. L. Organization, nucleation, and acetylation of microtubules in Xenopus laevis oocytes: a study by confocal immunofluorescence microscopy. Dev. Biol. 143, 346–362 (1991)

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to thank V. Korzh and C.-H. Hu for the plasmids for in situ hybridization, and K. Deen and M. Wyatt for editing the manuscript. This work was supported by grants from the National Science Council and Academia Sinica, Taiwan. Author Contributions H.-J.H. performed all the experiments; M.-R.L. synthesized F–P5; C.-T.C. devised and supervised the F–P5 synthesis scheme; B.-c.C. oversaw the execution of the entire project.

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Authors and Affiliations

  1. Institute of Molecular Biology, Academia Sinica, Nankang, 11529, Taipei, Taiwan

    Hwei-Jan Hsu & Bon-chu Chung

  2. Graduate Institute of Life Sciences, National Defense Medical Center, 11490, Taipei, Taiwan

    Hwei-Jan Hsu

  3. Department of Chemistry, National Taiwan University, 10617, Taipei, Taiwan

    Ming-Ren Liang & Chao-Tsen Chen

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  1. Hwei-Jan Hsu
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Correspondence to Bon-chu Chung.

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Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1–9 (PPT 7800 kb)

Supplementary Figure Legends

This file contains text to accompany the above Supplementary Figures. (DOC 37 kb)

Supplementary Table

Reducing cyp11a1 expression resulted in epibolic defect. (DOC 38 kb)

Supplementary Methods

This file details the methods for fish maintenance, morpholinos and analysis of genomic structures, plasmids, microinjection and detection of gene expression, steroid detection, RT-PCR, and synthesis of fluorescein conjugated pregnenolone. (DOC 215 kb)

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Hsu, HJ., Liang, MR., Chen, CT. et al. Pregnenolone stabilizes microtubules and promotes zebrafish embryonic cell movement. Nature 439, 480–483 (2006). https://doi.org/10.1038/nature04436

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