Abstract
Stimulus dependence is a general feature of developing animal sensory systems. In this respect, it has extensively been shown earlier that fish inner ear otoliths can act as test masses as their growth is strongly affected by altered gravity such as hypergravity obtained using centrifuges, by (real) microgravity achieved during spaceflight or by simulated microgravity using a ground-based facility. Since flight opportunities are scarce, ground-based simulators of microgravity, using a wide variety of physical principles, have been developed to overcome this shortcoming. Not all of them, however, are equally well suited to provide functional weightlessness from the perspective of the biosystem under evaluation. Therefore, the range of applicability of a particular simulator has to be extensively tested. Earlier, we have shown that a Rotating-Wall Vessel (RWV) can be used to provide simulated microgravity for developing Zebrafish regarding the effect of rotation on otolith development. In the present study, we wanted to find the most effective speed of rotation and identify the appropriate developmental stage of Zebrafish, where effects are the largest, in order to provide a methodological basis for future in-depth analyses dedicated to the physiological processes underlying otolith growth at altered gravity. Last not least, we compared data on the effect of simulated microgravity on the size versus the weight of otoliths, since the size usually is measured in related studies due to convenience, but the weight more accurately approximates the physical capacity of an otolith. Maintaining embryos at 10 hours post fertilization for three days in the RWV, we found that 15 revolutions per minute (rpm) yielded the strongest effects on otolith growth. Maintenance of Zebrafish staged at 10 hpf, 1 day post fertilization (dpf), 4 dpf, 7 dpf and 14 dpf for three days at 15 rpm resulted in the most prominent effects in 7 dpf larvae. Weighing versus measuring the size of otoliths yielded basically similar results, but the data gained by weighing were more distinct. Overall, our results clearly support the concept that the environmental gravity vector regulates fish otolith growth in terms of the pendulum model of otolith test masses, and that wall vessel rotation is a valuable means to provide functional weightlessness from the perspective of developing Zebrafish. We recommend that Zebrafish embryos staged 7 dpf (or possibly slightly elder) are rotated at 15 rpm in a Rotating-Wall Vessel as used in the present study for further experiments designed to elucidate the mechanisms underlying (altered gravity affected) otolith growth.
Similar content being viewed by others
Abbreviations
- dpf:
-
Days post fertilization
- hpf:
-
Hours post fertilization
- rpm:
-
Revolutions per minute
- RWV:
-
Rotating-Wall Vessel
- WVR:
-
Wall vessel rotation
References
Anken, R.: On the role of the central nervous system in regulating the mineralisation of inner-ear otoliths of fish. Protoplasma 229, 205–208 (2006)
Anken, R., Hilbig, R.: OMEGAHAB on FOTON-M3: Effect of long-term microgravity on the mineralisation of inner ear otoliths of fish. In: Proceedings of 4th China-German workshop on microgravity and space life sciences, p 58 (2009)
Anken, R., Rahmann, H.: Effect of altered gravity on the neurobiology of fish. Naturwissenschaften 86, 155–167 (1999)
Anken, R., Rahmann, H.: Gravitational zoology: How animals use and cope with gravity. In: Horneck, G., Baumstark-Khan, C. (eds.) Astrobiology, pp 315–333. Springer, Berlin (2002)
Anken, R., Ibsch, M., Breuer, J., Rahmann, H.: Effect of hypergravity on the Ca/Sr composition of developing otoliths of larval cichlid fish (Oreochromis mossambicus). Comp. Biochem. Physiol. A 128, 369–377 (2001)
Anken, R., Kappel, T., Rahmann, H.: Morphometry of fish inner ear otoliths after development at 3 g hypergravity. Acta Otolaryngol. (Stockh.) 118, 534–539 (1998)
Anken, R., Baur, U., Hilbig, R.: Clinorotation increases the growth of utricular otoliths of developing cichlid fish. Microgravity Sci. Technol. 22, 151–154 (2010)
Anken, R., Brungs, S., Grimm, D., Knie, M., Hilbig, R.: Fish inner ear otolith growth under real microgravity (spaceflight) and clinorotation. Microgravity Sci. Technol. 28(3), 351–356 (2016)
Bever, M., Fekete, M.: Atlas of the developing inner ear in zebrafish. Dev. Dyn. 223, 536–543 (2002)
Briegleb, W.: Some qualitative and quantitative aspects of the fast-rotating clinostat as a research tool. ASGSB Bull. 5, 23–30 (1992)
Brungs, S., Egli, M., Wuest, S., Christianen, P., van Loon, J., Ngo Anh, J., Hemmersbach, R.: Facilities for simulation of microgravity in the ESA ground-based facility programme. Microgravity Sci. Technol. 28, 191–203 (2016)
Brungs, S., Hauslage, J., Hilbig, R., Hemmersbach, R., Anken, R.: Effects of simulated microgravity on fish otolith growth: clinostat versus rotation- wall vessel. Adv. Space Res. 48, 792–798 (2011)
Colantonio, J., Vermot, J., Wu, D., Langenbacher, A., Fraser, S., Chen, J., Hill, K.: The dynein regulatory complex is required for ciliary motility and otolith biogenesis in the inner ear. Nature 457, 205–209 (2008)
de Vries, H.: The mechanics of the labyrinth otoliths. Acta Otolaryngol 38, 262–273 (1950)
Herranz, R., Anken, R., Boonstra, J., Braun, M., Christianen, P., de Geest, M., Hauslage, J., Hilbig, R., Hill, R., Lebert, M., Javier, F., Vagt, N., Ullrich, O., van Loon, J., Hemmersbach, R.: Ground-based facilities for simulation of microgravity: organism-specific recommendations for their use, and recommended terminology. Astrobiology 13, 1–17 (2013)
Kessler, J.: The internal dynamics of slowly rotating biological systems. ASGSB Bull. 5, 11–22 (1992)
Kondrachuk, A.: Models of the mechanical sensitivity and growth of otoliths in fish. J. Vest. Res. 13, 189–203 (2003)
Kondrachuk, A., Boyle, R.: Feedback hypothesis and the effects of altered gravity on formation and function of gravireceptors of mollusks and fish. Arch. Ital. Biol. 144, 75–87 (2006)
Li, X., Anken, R., Wang, G., Hilbig, R., Liu, Y.: Effects of sustained wall vessel rotation on the growth of larval Zebrafish inner ear otoliths. Microgravity Sci. Technol. 23, 13–18 (2011)
Moorman, S., Burress, C., Cordova, R., Slater, J.: Stimulus dependence of the development of the zebrafish (Danio rerio) vestibular system. J. Neurobiol. 38, 247–258 (1999)
Nüsslein-Volhard, C., Dahm, R.: Zebrafish. Oxford University Press (2002)
Pisam, M., Jammet, C., Laurent, D.: First steps of otolith formation of the zebrafish: Role of glycogen? Cell Tissue Res. 310, 163–168 (2002)
Riley, B., Zhu, C., Janetopoulos, C., Aufderheide, K.: A critical period of ear development controlled by distinct populations of ciliated cells in the zebrafish. Dev. Biol. 191, 191–201 (1997)
Riley, B., Moorman, S.: Development of utricular otoliths, but not saccular otoliths, is necessary for vestibular function and survival in the Zebrafish. Develop. Neurobiol. 43, 329–337 (2000)
Sebastian, C., Esseling, K., Horn, E.: Altered gravitational forces affect the development of the static vestibuloocular reflex in fish (Oreochromis mossambicus). J. Neurobiol. 46, 59–72 (2001)
Stooke-Vaughan, G., Huang, P., Hammond, K., Schier, A., Whitfield, T.: The role of hair cells, cilia and ciliary motility in otolith formation in the zebrafish otic vesicle. Development 139, 1777–1778 (2012)
Whitfield, T., Riley, B., Chiang, M., Bryan, P.: Development of the zebrafish inner ear. Dev. Dyn. 223, 427–458 (2002)
Wiederhold, M., Harrison, J., Parker, K., Nomura, H.: Otoliths developed in microgravity. J. Gravit. Physiol. 7, 39–42 (2000)
Wiederhold, M., Gao, W., Harrison, J., Parker, K.: Early development of gravity-sensing organ in microgravity. In: Buckey, J., Homick, J. (eds.) The Neurolab Spacelab Mission, NASA SP-2003-525, pp 123–132 (2003a)
Wiederhold, M., Harrison, J., Gao, W.: A critical period for gravitational effects on otolith formation. J. Vest. Res. 13, 205–214 (2003b)
Wu, D., Freund, B., Fraser, E., Vermot, J.: Mechanistic basis of otolith formation during teleost inner ear development. Dev. Cell 20, 271–278 (2011)
Acknowledgments
This work was supported by the Chinese Natural Science Foundation (31370421), the Project of Chinese Manned Spaceflight and the National Major Programs of Water Body Pollution Control and Remediation (2014ZX07302003-07).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare there are no conflicts of interest.
Rights and permissions
About this article
Cite this article
Li, X., Anken, R., Liu, L. et al. Effects of Simulated Microgravity on Otolith Growth of Larval Zebrafish using a Rotating-Wall Vessel: Appropriate Rotation Speed and Fish Developmental Stage. Microgravity Sci. Technol. 29, 1–8 (2017). https://doi.org/10.1007/s12217-016-9518-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12217-016-9518-5