Abstract
The mammalian oocyte develops within a complex of somatic cells known as a follicle, within which signals from the somatic cells regulate the oocyte, and signals from the oocyte regulate the somatic cells. Because isolation of the oocyte from the follicle disrupts these communication pathways, oocyte physiology is best studied within an intact follicle. Here we describe methods for quantitative microinjection of follicle-enclosed mouse oocytes, thus allowing the introduction of signaling molecules as well as optical probes into the oocyte within its physiological environment.
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References
Mehlmann, L.M., Jones, T.L.Z., and Jaffe, L.A. (2002) Meiotic arrest in the mouse follicle maintained by a Gs protein in the oocyte. Science 297, 1343–1345.
Mehlmann, L.M., Saeki, Y., Tanaka, S., Brennan, T.J., Evsikov, A.V., Pendola, F.L., Knowles, B.B., Eppig, J.J., and Jaffe, L.A. (2004) The Gs-linked receptor GPR3 maintains meiotic arrest in mammalian oocytes. Science 306, 1947–1950.
Mehlmann, L.M., Kalinowski, R.R., Ross, L.F., Hewlett, E.L., and Jaffe, L.A (2006). Meiotic resumption in response to luteinizing hormone is independent of a Gi family G protein or calcium in the mouse oocyte. Dev. Biol. 299, 345–355.
Kalinowski, R.R., Berlot, C.H., Jones, T.L.Z., Ross, L.F., Jaffe, L.A., and Mehlmann, L.M. (2004) Maintenance of meiotic prophase arrest in vertebrate oocytes by a Gs protein-mediated pathway. Dev. Biol. 267, 1–13.
Simon, A.M., Goodenough, D.A., Li, E., and Paul, D.L. (1997) Female infertility in mice lacking connexin 37. Nature 385, 525–529.
Freudzon, L., Norris, R.P., Hand, A.R., Tanaka, S., Saeki, Y., Jones, T.L.Z., Rasenick, M.M., Berlot, C.H., Mehlmann, L.M., and Jaffe, L.A. (2005) Regulation of meiotic prophase arrest in mouse oocytes by GPR3, a constitutive activator of the Gs G protein. J. Cell Biol. 171, 255–265.
Norris, R.P., Freudzon, L., Freudzon, M., Hand, A.R., Mehlmann, L.M., and Jaffe, L.A. (2007) A Gs-linked receptor maintains meiotic arrest in mouse oocytes, but luteinizing hormone does not cause meiotic resumption by terminating receptor-Gs signaling. Dev. Biol. 310, 240–249.
Mehlmann, L.M. (2005) Oocyte-specific expression of Gpr3 is required for the maintenance of meiotic arrest in mouse oocytes. Dev. Biol. 288, 397–404.
Eppig, J.J., Viveiros, M.M., Marin-Bivens, C., De La Fuente, R. (2004) Regulation of mammalian oocyte maturation, in The Ovary, 2nd edition (Leung, P.C.K. and Adashi, E.Y., ed.), Elsevier/Academic Press, San Diego, CA, pp. 113–129.
Mehlmann, L.M. (2005) Stops and starts in mammalian oocytes: recent advances in understanding the regulation of meiotic arrest and oocyte maturation. Reproduction 130, 791–799.
Nikolaev, V.O., and Lohse, M.J. (2006) Monitoring of cAMP synthesis and degradation in living cells. Physiology 21, 86–92.
Matzuk, M., Burns, K.H., Viveiros, M.M., and Eppig, J.J. (2002) Intercellular communication in the mammalian ovary: oocytes carry the conversation. Science 296, 2178–2180.
Park, J.Y., Su, Y.Q., Ariga, M., Law, E., Jin, S.L.C., and Conti, M. (2004) EGF-like growth factors as mediators of LH action in the ovulatory follicle. Science 303, 682–684.
Hiramoto, Y. (1962) Microinjection of the live spermatozoa into sea urchin eggs. Exp. Cell Res. 27, 416–426.
Kiehart, D.P. (1982) Microinjection of echinoderm eggs: apparatus and procedures. Meth. Cell Biol. 25, 13–31.
Jaffe, L.A., and Terasaki, M. (2004) Quantitative microinjection of oocytes, eggs, and embryos. Meth. Cell Biol. 74, 219–242.
Su, Y.-Q., Denegre, J.M., Wigglesworth, K., Pendola, F.L., O’Brien, M.J., and Eppig, J.J. (2003) Oocyte-dependent activation of mitogen-activated protein kinase (ERK1/2) in cumulus cells is required for the maturation of the mouse oocyte–cumulus cell complex. Dev. Biol. 263, 126–138.
Schuh, M., and Ellenberg, J. (2007) Self-organization of MTOCs replaces centrosome function during acentrosomal spindle assembly in live mouse oocytes. Cell 130, 484–498.
Kaila, K., and Voipio, J. (1985) A simple method for dry beveling of micropipettes used in the construction of ion-selective micro-electrodes. J. Physiol. 369, 8p.
Kline, D. (2009) Quantitative microinjection of mouse oocytes and eggs. Methods Mol. Biol. 518.
Acknowledgments
We thank John Eppig, Marilyn O'Brien, and Karen Wigglesworth for showing us how to obtain and culture mouse follicles, and Melina Schuh for critical reading of the manuscript. This work was supported by grants from the NIH.
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Jaffe, L.A., Norris, R.P., Freudzon, M., Ratzan, W.J., Mehlmann, L.M. (2009). Microinjection of Follicle-Enclosed Mouse Oocytes. In: Carroll, D. (eds) Microinjection. Methods in Molecular Biology, vol 518. Humana Press. https://doi.org/10.1007/978-1-59745-202-1_12
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DOI: https://doi.org/10.1007/978-1-59745-202-1_12
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