One of the hallmarks of fertilization is a transient increase in the levels of free calcium in the newly fertilized oocyte. This calcium """"""""spike"""""""" has been demonstrated during in vitro fertilization in a number of organisms. We are interested in whether Caenorhabditis elegans oocytes also exhibit a similar calcium spike at fertilization. Unlike other organisms, the C elegans oocyte can not be removed from the ovary and remain viable, therefore the techniques of in vitro fertilization are not available. We plan to circumvent this problem by imaging fertilization and early development in utero. To detect calcium transients we will inject calcium indicator dyes [Ca-green dextran (I OK) and/or Ca-crimson dextran (I OK), Molecular Probes, Inc., Eugene, OR] directly into C elegans obcytes. The worms with injected oocytes will be subsequently anesthetized in 0. 1% tricaine and 0.01 % tetramisole to immobilize them for the purpose of imaging. Ile anesthetic does not affect oviduct musculature, thus allowing fertilized oocytes to pass from the ovary to the uterus. The all-solid-state multi-photon imaging system developed at the MM will be used to image the oocytes in timelapse. The system is configured to collect simultaneously recorded images of both the fluorescence signal and the transmitted IR brightfield image. We will observe the changes in calcium levels in oocytes during the process of maturation prior to fertilization while the oocyte is resident in the ovary, during the process of fertilization as the oocyte passes through the spermatheca and during early development (meiosis and mitosis) of the embryo in the uterus.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
3P41RR000570-28S1
Application #
6117262
Study Section
Project Start
1998-09-30
Project End
2000-06-30
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
28
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Malecki, Marek; Putzer, Emily; Sabo, Chelsea et al. (2014) Directed cardiomyogenesis of autologous human induced pluripotent stem cells recruited to infarcted myocardium with bioengineered antibodies. Mol Cell Ther 2:
Malecki, Marek (2014) 'Above all, do no harm': safeguarding pluripotent stem cell therapy against iatrogenic tumorigenesis. Stem Cell Res Ther 5:73
Mavroudi, Maria; Zarogoulidis, Paul; Porpodis, Konstantinos et al. (2014) Stem cells' guided gene therapy of cancer: New frontier in personalized and targeted therapy. J Cancer Res Ther (Manch) 2:22-33
Malecki, Marek; Dahlke, Jessica; Haig, Melissa et al. (2013) Eradication of Human Ovarian Cancer Cells by Transgenic Expression of Recombinant DNASE1, DNASE1L3, DNASE2, and DFFB Controlled by EGFR Promoter: Novel Strategy for Targeted Therapy of Cancer. J Genet Syndr Gene Ther 4:152
Zarogoulidis, Paul; Darwiche, Kaid; Sakkas, Antonios et al. (2013) Suicide Gene Therapy for Cancer - Current Strategies. J Genet Syndr Gene Ther 4:
Malecki, Marek; Sabo, Chelsea; Putzer, Emily et al. (2013) Recruitment and retention of human autologous CD34+ CD117+ CD133+ bone marrow stem cells to infarcted myocardium followed by directed vasculogenesis: Novel strategy for cardiac regeneration. Mol Cell Ther 1:
Malecki, Marek; LaVanne, Christine; Alhambra, Dominique et al. (2013) Safeguarding Stem Cell-Based Regenerative Therapy against Iatrogenic Cancerogenesis: Transgenic Expression of DNASE1, DNASE1L3, DNASE2, DFFB Controlled By POLA1 Promoter in Proliferating and Directed Differentiation Resisting Human Autologous Pluripotent J Stem Cell Res Ther Suppl 9:
Malecki, Marek; Tombokan, Xenia; Anderson, Mark et al. (2013) TRA-1-60(+), SSEA-4(+), POU5F1(+), SOX2(+), NANOG(+) Clones of Pluripotent Stem Cells in the Embryonal Carcinomas of the Testes. J Stem Cell Res Ther 3:
Malecki, Marek (2013) Improved targeting and enhanced retention of the human, autologous, fibroblast-derived, induced, pluripotent stem cells to the sarcomeres of the infarcted myocardium with the aid of the bioengineered, heterospecific, tetravalent antibodies. J Stem Cell Res Ther 3:
Malecki, Marek; Malecki, Bianca (2012) Routing of Biomolecules and Transgenes' Vectors in Nuclei of Oocytes. J Fertili In Vitro 2012:108-118

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