Monika A. Ward
	Affiliation: School of Medicine, Institute for Biogenesis Research Position: Professor Degree: MS, PhD (Poznan University, Poland) Phone: (808) 956 0779 Fax: (808) 956 7316 Email: mward@hawaii.edu Address: 1960 East-West Rd, Room E104, Honolulu, HI 96822

Sperm Genetics and Function in Fertilization

Assisted reproduction enables achieving fertilization when normal conception does not occur due to a variety of gamete defects. Intracytoplasmic sperm injection (ICSI) is the injection of a single spermatozoon directly into the cytoplasm of an oocyte using an injection pipette. This technique has been applied successfully in the treatment of infertile couples and is now widely used as a method of human assisted reproduction (ART). In addition to its obvious role in overcoming the infertility, ICSI is also an excellent tool allowing exploring new venues of reproductive biology. It provides a unique opportunity to examine the mechanisms underlying male infertility by looking into the reproductive potentials of individual spermatozoa.

The primary research interest of the lab is to study sperm genetics and function in fertilization in the context of assisted reproduction, utilizing advanced techniques of gamete and embryo micromanipulation combined with cytogenetic and molecular biology techniques, in a mouse model. The overall goal is to explore how the 'genetic composition' of sperm translates on its function in fertilization.

The ongoing projects involve:

(1) Reproducing subfertile and infertile mice with phenotypes that mimic various human male infertility syndromes to test for ART effects;

(2) Studying sperm DNA damage, its origin and consequences for fertilization and embryo development;

(3) Examining the function of the Y chromosome encoded genes in male fertility;

(4) Identifying and characterizing the mechanism of action of the Y chromosome encoded susceptibility locus mechanistically relevant to multiple sclerosis.

Figure 1. Teratozoospermia is a fertility defect expressed as an increased proportion of morphologically abnormal sperm that affects both humans and mice. Morphologically abnormal sperm (A) are often unable to fertilize oocytes rendering males infertile. Infertility can be overcome with intracytoplasmic sperm injection (ICSI), a method that allows injecting a spermatozoon directly into the oocyte using an injection pipette (B). The resulting embryos can be tested for their potential to develop in vitro from zygote to blastocyst stage (C), their genetic integrity evidenced by analysis of paternal or maternal chromosome complement in the zygote (D) and their potential to develop in vivo to live offspring after embryo transfer (E). Such analyses allow testing sperm function in infertile males with teratozoospermia, and can also be applied to other infertility defects.

	Yasuhiro Yamauchi Junior Researcher yyamauch@hawaii.edu Lab 956-5556 Office 956-5475		Jonathan M. Riel Graduate Student Jonathan.Riel@gmail.com Office 956-5556 Lab 956-9761
	Victor Ruthig Graduate Student vruthig@hawaii.edu Office 956-5556 Lab 956-9761		Egle Ortega Graduate Student egle@hawaii.edu Office 956-5556 Lab 956-9761

• Szczygiel M., Kurpisz M. (2001) Chromosomal anomalies in human gametes and pre-implantation embyos, and their potential effect on reproduction. Andrologia 33: 294-265.
• Kusakabe, H., Szczygiel, M.A., Whittingham, D.G. and Yanagimachi, R. (2001) Maintenance of genetic integrity in frozen and freeze-dried mouse spermatozoa. Proceedings of the National Academy of Sciences USA 98:13501-13506.
• Szczygiel, M.A. and Ward, W.S. (2002) The combination of dithiothreitol and detergent treatment of spermatozoa causes paternal chromosomal damage. Biology of Reproduction 67:1532-1537.
• Mohar, I., Szczygiel, M.A., Yanagimachi, R. and Ward, W.S. (2002) Sperm nuclear halos can transform into normal chromosomes after injection into oocytes. Molecular Reproduction and Development 62:416-420.
• Szczygiel, M.A., Kusakabe, H., Yanagimachi, R. and Whittingham, D.G. (2002) Separation of motile populations of spermatozoa prior to freezing is beneficial for subsequent fertilization in vitro: A study with various mouse strains. Biology of Reproduction 67:287-292.
• Szczygiel, M.A., Kusakabe, H., Yanagimachi, R. and Whittingham, D.G. (2002) Intracytoplasmic sperm injection is more efficient than in vitro fertilization for generating mouse embryos from cryopreserved spermatozoa. Biology of Reproduction 67:1278-1284.
• Szczygiel, M.A., Moisyadi, S. and Ward, W.S. (2003) Expression of foreign DNA is associated with paternal chromosome degradation in ICSI-mediated transgenesis. Biology of Reproduction 68:1903-1910.
• Ward, M.A., Kaneko, T., Kusakabe, H., Biggers, J.D., Whittingham, D.G. and Yanagimachi, R. (2003) Long term preservation of mouse spermatozoa after freeze-drying and freezing without cryoprotection. Biology of Reproduction 69:2100-2108.
• Ward, M.A. and Ward, W.S. (2004) Functional aspects of mammalian sperm chromatic structure. Reproduction Fertility and Development 16:547-554.
• Ward, M.A. (2005) ICSI effects in infertile azh mutant mice. Biology of Reproduction 73:193-200.
• Ward, M.A. and Burgoyne, P.S. (2006) The effects of deletions of the mouse Y chromosome long arm on sperm function - intracytoplasmic sperm injection (ICSI)-based analysis. Biology of Reproduction 74:652-658.
• Ajduk A., Yamauchi Y., Ward M.A. (2006) Sperm chromatin remodeling after intracytoplasmic sperm injection differs from that of in vitro fertilization. Biology of Reproduction 75: 442-451.
• Yamauchi, Y. and Ward M.A. (2007) Preservation of ejaculated mouse spermatozoa from fertile C57BL/6 and infertile Hook1/Hook1 mice collected from the uteri of mated females. Biology of Reproduction 76: 1002-1008.
• Riel J.M., Huang T.F. and Ward M.A. (2007) Freezing-free preservation of human spermatozoa – a pilot study. Archives of Andrology, 53: 275-284.
• Yamauchi, Y., Ajduk, A, Riel J and Ward M.A. (2007) Ejaculated and epididymal mouse spermatozoa are different in their susceptibility to nuclease-dependent DNA damage and in their nuclease activity. Biology of Reproduction 77: 636-47.
• Yamauchi, Y., Doe B., Ajduk A., Ward M.A. (2007) Genomic DNA damage in mouse transgenesis. Biology of Reproduction 77: 803-812.
• Yamauchi, Y., Ward, M.A., Ward, W.S. (2009) Asynchronous DNA replication and origin licensing in the mouse one-cell embryo. Journal of Cellular Biochemistry 107: 214-223.
• Collier, A.C., Miyagi, S.J., Yamauchi, Y. and Ward, M.A. (2009) Assisted reproduction technologies impair placental steroid metabolism. Journal of Steroid Biochemistry and Molecular Biology 116: 21-28.
• Yamauchi, Y., Riel, J.M., Wong, S. J., Ojarikre, O.A., Burgoyne, P.S. and Ward, M.A. (2009) Live offspring from mice lacking the Y chromosome long arm gene complement. Biology of Reproduction 81: 353-361.
• Cocquet, J., Ellis, P. J.I., Yamauchi, Y., Mahadevaiah, S.K., Affara, N.A., Ward, M.A., Burgoyne, P.S. (2009) The multicopy gene Sly represses the sex chromosomes in the male mouse germline after meiosis. PLOS Biology 7(11)
• Cocquet, J., Ellis, P.J., Yamauchi, Y., Riel, J.M., Karacs, T.P., Rattigan, A., Ojarikre, O.A., Affara, N.A., Ward, M.A., Burgoyne, P.S. (2010) Deficiency in the multicopy Sycp3-like X-linked genes Slx and Slxl1 causes major defects in spermatid differentiation. Molecular Biology of the Cell. 21: 3497-3505.
• Jefferey M. Raunig, Yasuhiro Yamauchi, Monika A. Ward and Abby C. Collier (2011): Assisted Reproduction Technologies Alter Steroid Delivery to the Mouse Fetus During Pregnancy. Journal of Steroid Biochemistry and Molecular Biology. August. 126 (1-2): 26-34. PMCID: PMC3114261
• Yamauchi, Y., Riel, J.M., Ward, M.A. (2011): Paternal DNA damage resulting from various sperm treatments persists after fertilization and is similar prior and after DNA replication. Journal of Andrology May 5. [Epub ahead of print] NIHMSID: NIHMS368852
• Riel, J.M., Yamauchi, Y., Huang, T.T.F., Grove, J., Ward, M.A. (2011) Short-term storage of human spermatozoa in electrolyte-free medium without freezing maintains sperm genetic integrity better than cryopreservation. Biology of Reproduction 85(3): 536-47. Epub 2011 May 18 PMCID: PMC3159537
• Sugawara, A., Pearson, B.L., Blanchard, D.C., Ward, M.A. (2011) Mouse females devoid of exposure to males during fetal development exhibit increased maternal behavior. Psychoneuroendocrinology. 37: 383-395 [Epub ahead of print Jul 29, 2011] PMCID: PMC3212613
• Raunig, J.M., Yamauchi, Y., Ward, M.A., Collier, A.C. (2011) Placental inflamation and oxidative stress in the mouse model of assisted reproduction. Placenta 11: 852-858 [Epub ahead of print Sep 1] PMCID: PMC3210407
• Abby C. Collier, Kristen A. Milam, Atsushi Sugawara, Yasuhiro Yamauchi, Monika A. Ward (2011): Upregulation of Ugt1a genes in placentas and fetal livers in a murine model of assisted reproduction. Placenta 33: 77-80 [Epub ahead of print Nov 23, 2011] PMCID: PMC3249018
• Ward M. A. (2012): Wild-derived inbred mice no longer ART-resistant. Biology of Reproduction 86:168, 1-2 (Epub Mar 14, 2012). PMCID: PMC3364932.
• Sugawara A, Sato B, Bal E, Collier A.C., Ward M. A. (2012): Blastomere removal from cleavage-stage mouse embryos alters steroid metabolism during pregnancy. Biology of Reproduction 87:1-9 (Epub Jul 5, 2012). PMCID: PMC3406553.
• Sugawara A. and Ward M. A.(2013): Biopsy of embryos produced by in vitro fertilization affects development in C57BL/6 mouse strain. Theriogenology. 79: 234-241(Epub ahead of print Nov 19, 2012). PMCID: PMC3534949
• Riel J. M., Yamauchi Y, Sugawara A, Li H. Y. L., Ruthig V, Stoytcheva Z, Ellis P.J.I., Cocquet J., Ward M. A. (2012): Deficiency of the multi-copy mouse Y gene Sly causes sperm DNA damage and abnormal chromatin packaging. Journal of Cell Science (Epub Nov 23 2012)
• Gawecka J. E., Marh J., Ortega M., Yamauchi Y., Ward M. A., Ward W. S. (2013): Mouse Zygotes Respond to Severe Sperm DNA Damage by Delaying Paternal DNA Replication and Embryonic Development. PLoS One 8(2):e56385 (Epub Feb 19, 2013) PMCID: PMC3576397
• Yamauchi Y., Riel Jonathan M., Stoytcheva Z., Ward Monika A (2014): Two Y genes can replace the entire Y chromosome for assisted reproduction in the mouse. Science 2014 Jan 3;343(6166):69-72. doi: 10.1126/science.1242544. Epub 2013 Nov 21.PMID:24263135.
• Vernet N, Mahadevaiah SK, Yamauchi Y, Decarpentrie F, Mitchell MJ, Ward MA, Burgoyne PS (2014): Mouse Y-linked Zfy1 and Zfy2 are expressed during the male-specific interphase between meiosis I and meiosis II and promote the 2nd meiotic division. PLoS Genet. 2014 Jun 26;10(6):e1004444.
• Sato BL, Sugawara A, Ward MA, Collier AC (2014): Single blastomere removal from murine embryos is associated with activation of matrix metalloproteinases and Janus kinase/Signal Transducers and Activators of Transcription pathways of placental inflammation. Mol Hum Reprod. 2014 Sep 1.
• Collier AC, Yamauchi Y, Sato BL, Rougee LR, Ward MA (2014): UDP-glucuronosyltransferase (Ugt) 1a Enzymes are Present and Active in the Mouse Blastocyst. Drug Metab Dispos. 2014 Sep 8
• Comish P, Lai L, Yamauchi Y, Weng C, Gunapala S, Naff K, Ward MA, Meistrich M (2014): Increasing ambient tempratures by exposure to elevated ambient temperatures restores spermatogonesis in adult Utp14b-jsd mutant (jsd) mice. Andrology (Epub Oct 90, 2014) 2015 Mar;3(2):376-84. PMCID: PMC4391977 [Available on 2016-03-01].
• Sato B.L.M, Sugawara A., Ward M.A, Collier A.C. (2014): Single blastomere removal from murine embryos is associated with activation of MMP and Jak/STAT pathways of placental inflammation. Molecular Human Reproduction (Epub Sep 1, 2014) Dec; 20(12): 1247-57. PMCID: PMC4235574
• Ortega E., Ruthig V., Ward M.A. (2015): Sry-independent overexpression of Sox9 supports spermatogenesis and fertility in the mouse. Biology in Reproduction (Epub Nov 4, 2015). Dec;93(6):141. PMID: 26536904 [PubMed - in process]
• Yamauchi Y., Riel J.M., Ruthig V., Ward M.A (2015): Mouse Y-encoded transcription factor Zfy2 is essential for sperm formation and function in assisted fertilization. PLoS Genetics 11(12): e1005476. [PubMed - in process]
• Yamauchi Y., Riel, J.M., Ruthig V.A., Ortega E.A., Mitchell J.M., Ward, M.A. (2016): Two genes substitute for the mouse Y chromosome for spermatogenesis and reproduction. Science, 29 Jan 2016, Vol 351 Issue 6272, pp 514-516, Download Reprint PDF