Hostname: page-component-78c5997874-dh8gc Total loading time: 0 Render date: 2024-11-10T13:52:00.288Z Has data issue: false hasContentIssue false
  • English
  • Français

FACS selection of valuable mutant mouse round spermatids and strain rescue via round spermatid injection

Published online by Cambridge University Press:  20 December 2013

Lian Zhu ,
Wei Zhou ,
Peng-Cheng Kong ,
Mei-Shan Wang ,
Yan Zhu ,
Li-Xin Feng ,
Xue-Jin Chen  and
Man-Xi Jiang
Lian Zhu
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Wei Zhou
Affiliation:
Department of Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Peng-Cheng Kong
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Mei-Shan Wang
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. College of Wildlife Resource, Northeast Forestry University, Harbin 150040, China.
Yan Zhu
Affiliation:
Key Laboratory of Contraceptive Drugs and Devices of National Population and Family Planning Committee, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
Li-Xin Feng
Affiliation:
Department of Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Xue-Jin Chen*
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Man-Xi Jiang*
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
*
All correspondence to: Man-Xi Jiang or Xue-Jin Chen. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Tel: +86 21 63846590, ext. 776539. e-mail: manxijiang@yahoo.com or chenxuejin@shsmu.edu.cn
All correspondence to: Man-Xi Jiang or Xue-Jin Chen. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Tel: +86 21 63846590, ext. 776539. e-mail: manxijiang@yahoo.com or chenxuejin@shsmu.edu.cn
Get access Rights & Permissions [Opens in a new window]

Summary

Round spermatid injection (ROSI) into mammalian oocytes can result in the development of viable embryos and offspring. One current limitation to this technique is the identification of suitable round spermatids. In the current paper, round spermatids were selected from testicular cells with phase contrast microscopy (PCM) and fluorescence-activated cell sorting (FACS), and ROSI was performed in two strains of mice. The rates of fertilization, embryonic development and offspring achieved were the same in all strains. Significantly, round spermatids selected by PCM and FACS were effectively used to rescue the infertile Pten-null mouse. The current results indicate that FACS selection of round spermatids can not only provide high-purity and viable round spermatids for use in ROSI, but also has no harmful effects on the developmental capacity of subsequently fertilized embryos. It was concluded that round spermatids selected by FACS are useful for mouse strain rederivation and rescue of infertile males; ROSI should be considered as a powerful addition to the armamentarium of assisted reproduction techniques applicable in the mouse.

Keywords

MouseFACSROSIRound spermatidsStrain rescue
Type
Research Article
Information
Zygote , Volume 23 , Issue 3 , June 2015 , pp. 336 - 341
Copyright
Copyright © Cambridge University Press 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Barchi, M., Geremia, R., Magliozzi, R. & Bianchi, E. (2009). Isolation and analyses of enriched populations of male mouse germ cells by sedimentation velocity: the centrifugal elutriation. Methods Mol. Biol. 558, 299–321.CrossRefGoogle ScholarPubMed
Bos-Mikich, A., Swann, K. & Whittingham, D.G. (1995). Calcium oscillations and protein synthesis inhibition synergistically activate mouse oocytes. Mol. Reprod. Dev. 41, 84–90.Google Scholar
Chang, Y.F., Lee-Chang, J.S., Panneerdoss, S., MacLean, J.A. 2nd & Rao, M.K. (2011). Isolation of Sertoli, Leydig, and spermatogenic cells from the mouse testis. Biotechniques 51, 341–4.Google Scholar
Getun, I.V., Torres, B. & Bois, P.R. (2011). Flow cytometry purification of mouse meiotic cells. J. Vis. Exp. 50, e2602. doi:10.3791 /2602 Google Scholar
Goodell, M.A. (2005). Stem cell identification and sorting using the Hoechst 33342 side population (SP.) In Current Protocols in Cytometry. New York: John Wiley & Sons, Inc. Chapter 9, Unit 9.18.Google Scholar
Hikichi, T., Kishigami, S., Thuan, N.V., Ohta, H., Mizutani, E., Wakayama, S. & Wakayama, T. (2005). Round spermatids stained with MitoTracker can be used to produce offspring more simply. Zygote 13, 55–61.Google Scholar
Hirabayashi, M., Kato, M., Aoto, T., Ueda, M. & Hochi, S. (2002). Rescue of infertile transgenic rat lines by intracytoplasmic injection of cryopreserved round spermatids. Mol. Reprod. Dev. 62, 295–9.CrossRefGoogle ScholarPubMed
Kimura, Y. & Yanagimachi, R. (1995a). Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring. Development 121, 2397–405.CrossRefGoogle ScholarPubMed
Kimura, Y. & Yanagimachi, R. (1995b). Development of normal mice from oocytes injected with secondary spermatocyte nuclei. Biol. Reprod. 53, 855–62.CrossRefGoogle ScholarPubMed
La Salle, S., Sun, F. & Handel, M.A. (2009). Isolation and short-term culture of mouse spermatocytes for analysis of meiosis. Methods Mol. Biol. 558, 279–97.CrossRefGoogle ScholarPubMed
Lassalle, B., Ziyyat, A., Testart, J., Finaz, C. & Lefevre, A. (1999). Flow cytometric method to isolate round spermatids from mouse testis. Hum. Reprod. 14, 388–94.CrossRefGoogle ScholarPubMed
Meng, X., Akutsu, H., Schoene, K., Reifsteck, C., Fox, E.P., Olson, S., Sariola, H., Yanagimachi, R. & Baetscher, M. (2002). Transgene insertion induced dominant male sterility and rescue of male fertility using round spermatid injection. Biol. Reprod. 66, 726–34.CrossRefGoogle ScholarPubMed
Morena, A.R., Boitani, C., Pesce, M., De Felici, M. & Stefanini, M. (1996). Isolation of highly purified type A spermatogonia from prepubertal rat testis. J. Androl. 17, 708–17.Google Scholar
Ogura, A., Matsuda, J. & Yanagimachi, R. (1994). Birth of normal young after electrofusion of mouse oocytes with round spermatids. Proc. Natl. Acad. Sci. USA 91, 7460–2.Google Scholar
Ogura, A., Ogonuki, N., Takano, K. & Inoue, K. (2001). Microinsemination, nuclear transfer, and cytoplasmic transfer: the application of new reproductive engineering techniques to mouse genetics. Mamm. Genome 12, 803–12.Google Scholar
Palermo, G., Joris, H., Devroey, P. & Van Steirteghem, A.C. (1992). Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet 340, 17–8.CrossRefGoogle ScholarPubMed
Parrilla, I., Vazquez, J.M., Cuello, C., Gil, M.A., Roca, J., Di Berardino, D. & Martinez, E.A. (2004). Hoechst 33342 stain and u.v. laser exposure do not induce genotoxic effects in flow-sorted boar spermatozoa. Reproduction 128, 615–21.Google Scholar
Pelengaris, S.A. & Moore, H.D. (1995). Separation of round spermatids from the rat using an immunoselection panning technique. Mol. Reprod. Dev. 41, 348–54.Google Scholar
Romero, Y., Meikar, O., Papaioannou, M.D., Conne, B., Grey, C., Weier, M., Pralong, F., De Massy, B., Kaessmann, H., Vassalli, J.D., Kotaja, N. & Nef, S. (2011). Dicer1 depletion in male germ cells leads to infertility due to cumulative meiotic and spermiogenic defects. PLoS One 6, e25241.Google Scholar
Sasagawa, I. & Yanagimachi, R. (1997). Spermatids from mice after cryptorchid and reversal operations can initiate normal embryo development. J. Androl. 18, 203–9.Google Scholar
Sasagawa, I., Kuretake, S., Eppig, J.J. & Yanagimachi, R. (1998). Mouse primary spermatocytes can complete two meiotic divisions within the oocyte cytoplasm. Biol. Reprod. 58, 248–54.Google Scholar
Sharan, S.K., Pyle, A., Coppola, V., Babus, J., Swaminathan, S., Benedict, J., Swing, D., Martin, B.K., Tessarollo, L., Evans, J.P., Flaws, J.A. & Handel, M.A. (2004). BRCA2 deficiency in mice leads to meiotic impairment and infertility. Development 131, 131–42.CrossRefGoogle ScholarPubMed
Sofikitis, N.V., Miyagawa, I., Agapitos, E., Pasyianos, P., Toda, T., Hellstrom, W.J. & Kawamura, H. (1994). Reproductive capacity of the nucleus of the male gamete after completion of meiosis. J. Assist. Reprod. Genet. 11, 335–41.Google Scholar
Sutovsky, P., Ramalho-Santos, J., Moreno, R.D., Oko, R., Hewitson, L. & Schatten, G. (1999). On-stage selection of single round spermatids using a vital, mitochondrion-specific fluorescent probe MitoTracker™ and high resolution differential interference contrast microscopy. Hum. Reprod. 14, 2301–12.Google Scholar
Tesarik, J., Mendoza, C. & Testart, J. (1995). Viable embryos from injection of round spermatids into oocytes. N. Engl. J. Med. 333, 525.Google Scholar
Van Steirteghem, A.C., Nagy, Z., Joris, H., Liu, J., Staessen, C., Smitz, J., Wisanto, A. & Devroey, P. (1993). High fertilization and implantation rates after intracytoplasmic sperm injection. Hum. Reprod. 8, 1061–6.Google Scholar
Wykes, S.M. & Krawetz, S.A. (2003). Separation of spermatogenic cells from adult transgenic mouse testes using unit-gravity sedimentation. Mol. Biotechnol. 25, 131–8.Google Scholar
Yanagimachi, R. (2001). Gamete manipulation for development: new methods for conception. Reprod. Fertil. Dev. 13, 3–14.CrossRefGoogle ScholarPubMed