Microinjection in Zebrafish Otocytes

Researchers at the University of Michigan are using WPI's PV820 for injecting a morpholino solution into the lumen of the otic vesicle of 1-day old zebrafish embryos.  Then, they use electroporation to introduce mif and mif-like morpholinos into the developing inner ear tissues.

Microinjection in Zebrafish Oocytes

Check out the JOVE video to see:

  • How to make electrodes for electroporation
  • How to setup the electroporation
  • How to inject the morpholinos
  • Examine some of the results

More Info

 

References

JOVE video: Holmes, K. E., Wyatt, M. J., Shen, Y., Thompson, D. A., Barald, K. F. "Direct Delivery of MIF Morpholinos Into the Zebrafish Otocyst by Injection and Electroporation Affects Inner Ear Development". J. Vis. Exp. (47), e2466, doi:10.3791/2466 (2011).

Warmerdam, T., Schröder, F., Wit, H., & Albers, F. (n.d.). Perilymphatic and endolymphatic pressures during endolymphatic hydrops. European Archives of Oto-Rhino-Laryngology, 260(1), 9–11. http://doi.org/10.1007/s00405-002-0518-2 

Wei, J., Song, J., Jiang, S., Zhang, G., Wheeler, D., Zhang, J., … Liu, R. (2017). Role of intratubular pressure during the ischemic phase in acute kidney injury. American Journal of Physiology - Renal Physiology, 312(6), F1158–F1165. http://doi.org/10.1152/ajprenal.00527.2016 

Petrie, R. J., Koo, H., & Yamada, K. M. (2014). Generation of compartmentalized pressure by a nuclear piston governs cell motility in a 3D matrix. Science, 345(6200), 1062–1065. http://doi.org/10.1126/science.1256965 

Petrie, R. J., Koo, H., & Yamada, K. M. (2014). Generation of compartmentalized pressure by a nuclear piston governs cell motility in a 3D matrix. Science (New York, N.Y.), 345(6200), 1062–5. http://doi.org/10.1126/science.1256965 

Petrie, R. J., & Koo, H. (2014). Direct measurement of intracellular pressure. Current Protocols in Cell Biology / Editorial Board, Juan S. Bonifacino ... [et Al.], 63, 12.9.1-9. http://doi.org/10.1002/0471143030.cb1209s63 

Pacella, J. J., Kameneva, M. V, Brands, J., Lipowsky, H. H., Vink, H., Lavery, L. L., & Villanueva, F. S. (2012). Modulation of pre-capillary arteriolar pressure with drag-reducing polymers: a novel method for enhancing microvascular perfusion. Microcirculation (New York, N.Y. : 1994), 19(7), 580–5. http://doi.org/10.1111/j.1549-8719.2012.00190.x 
Park, J. J.-H., Boeven, J. J., Vogel, S., Leonhardt, S., Wit, H. P., & Westhofen, M. (2012). Hydrostatic fluid pressure in the vestibular organ of the guinea pig. European Archives of Oto-Rhino-Laryngology, 269(7), 1755–1758. http://doi.org/10.1007/s00405-011-1813-6 

Hepatic microvascular pressure during anaphylactic shock in anesthetized rats. (2009). Microvascular Research, 78(2), 169–173. http://doi.org/10.1016/J.MVR.2009.06.007 

Valk, W. L., Wit, H. P., & Albers, F. W. J. (2006). Rupture of Reissner’s membrane during acute endolymphatic hydrops in the guinea pig: a model for Ménière’s disease? Acta Oto-Laryngologica, 126(10), 1030–1035. http://doi.org/10.1080/00016480600621722 

Kopp, R., Schwerte, T., & Pelster, B. (2005). Cardiac performance in the zebrafish breakdance mutant. The Journal of Experimental Biology, 208(Pt 11), 2123–34. http://doi.org/10.1242/jeb.01620 

Lucitti, J. L., Tobita, K., & Keller, B. B. (2005). Arterial hemodynamics and mechanical properties after circulatory intervention in the chick embryo. The Journal of Experimental Biology, 208, 1877–1885. http://doi.org/10.1242/jeb.01574 

Understanding cardiovascular physiology in zebrafish and Xenopus larvae: the use of microtechniques. (2003). Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 135(1), 131–145. http://doi.org/10.1016/S1095-6433(03)00044-8 

Hu, N., Yost, H. J., & Clark, E. B. (2001). Cardiac morphology and blood pressure in the adult zebrafish. The Anatomical Record, 264(1), 1–12. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/11505366 

Ishii, T., Kuwaki, T., Masuda, Y., & Fukuda, Y. (2001). Postnatal development of blood pressure and baroreflex in mice. Autonomic Neuroscience : Basic & Clinical, 94(1–2), 34–41. http://doi.org/10.1016/S1566-0702(01)00339-3 

Hu, N., Sedmera, D., Yost, H. J., & Clark, E. B. (2000). Structure and function of the developing zebrafish heart. The Anatomical Record, 260(2), 148–57. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/10993952 

Kelly, S. M., & Macklem, P. T. (1991). Direct measurement of intracellular pressure. The American Journal of Physiology, 260(June), C652–C657. http://doi.org/10.1002/0471143030.cb1209s63 

Proximodistal gradient in endoneurial fluid pressure. (1988). Experimental Neurology, 102(3), 368–370. http://doi.org/10.1016/0014-4886(88)90233-6 

Tanner, C., Frambach, D. A., & Misfeldt, D. S. (1983). Transepithelial transport in cell culture. A theoretical and experimental analysis of the biophysical properties of domes. Biophysical Journal, 43(2), 183–90. http://doi.org/10.1016/S0006-3495(83)84339-2 

Rabito, C. A., Tchao, R., Valentich, J., & Leighton, J. (1980). Effect of cell-substratum interaction on hemicyst formation by MDCK cells. In Vitro, 16(6), 461–8. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/6248454