This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Tsunoda, Y. Articles by Uchida, T. Search for Related Content PubMed PubMed Citation Articles by Tsunoda, Y. Articles by Uchida, T.

Differential sensitivity of mouse pronuclei and zygote cytoplasm to Hoechst staining and ultraviolet irradiation

Summary. The exposure of mouse zygotes pre-stained with Hoechst 33342 to u.v. irradiation for 20–30 sec significantly or completely inhibited development to blastocysts in vitro. However, development to the blastocyst stage of enucleated eggs receiving pronuclei from untreated eggs was as good as that of control reconstituted eggs when the cytoplasm originated from eggs exposed to u.v. irradiation for 20–30 sec, but was significantly lower when the cytoplasm was from eggs exposed for 40 sec. The chromosomes at the second metaphase stage could be removed with 15 sec of exposure to u.v. irradiation under a fluorescence microscope. Most eggs enucleated at the second metaphase that received a single inner cell mass nucleus (75%) showed pronuclear formation 6 h after activation; 23% of them developed to morphologically normal 2-cell eggs and 5% developed to blastocysts.

These results demonstrate that the cytoplasm of mouse zygotes is more resistant to u.v. irradiation after Hoechst staining. Eggs at the second metaphase, from which chromosomes have been removed under a fluorescence microscope, can therefore be used as cytoplasm recipients for nuclear transplantation of inner cell mass nuclei.

Keywords: u.v. irradiation; Hoechst staining; nuclear transplantation; mouse egg

This article has been cited by other articles:

				J.-W. Lee, S.-C. Wu, X. C. Tian, M. Barber, T. Hoagland, J. Riesen, K.-H. Lee, C.-F. Tu, W. T.K. Cheng, and X. Yang Production of Cloned Pigs by Whole-Cell Intracytoplasmic Microinjection Biol Reprod, September 1, 2003; 69(3): 995 - 1001. [Abstract] [Full Text] [PDF]

				V. Ivakhnenko, J. Cieslak, and Y. Verlinsky A microsurgical technique for enucleation of multipronuclear human zygotes Hum. Reprod., April 1, 2000; 15(4): 911 - 916. [Abstract] [Full Text] [PDF]

				V. Bordignon and L. C. Smith Ultraviolet-Irradiated Spermatozoa Activate Oocytes but Arrest Preimplantation Development After Fertilization and Nuclear Transplantation in Cattle Biol Reprod, December 1, 1999; 61(6): 1513 - 1520. [Abstract] [Full Text]

				H. Liu, C.-W. Wang, J. A. Grifo, L. C. Krey, and J. Zhang Reconstruction of mouse oocytes by germinal vesicle transfer: maturity of host oocyte cytoplasm determines meiosis Hum. Reprod., September 1, 1999; 14(9): 2357 - 2361. [Abstract] [Full Text] [PDF]

				Y. Verlinsky and S. Evsikov Karyotyping of human oocytes by chromosomal analysis of the second polar bodies Mol. Hum. Reprod., February 1, 1999; 5(2): 89 - 95. [Abstract] [Full Text] [PDF]

				Y Kato and Y Tsunoda Germ cell nuclei of male fetal mice can support development of chimeras to midgestation following serial transplantation Development, January 3, 1995; 121(3): 779 - 783. [Abstract] [PDF]