Abstract
Understanding the cellular events during fertilization in mammals is a major challenge that can contribute to the improvement of future infertility treatments in humans and reproductive performance in farm animals. Of special interest is the role of the oocyte and sperm cytoskeleton during the initial interaction between gametes. The aim of this chapter is to describe methods for studying cytoskeletal features during in vitro fertilization after intracytoplasmic sperm injection (ICSI) in humans. The following protocols will provide a detailed description of how to perform immunodetection and imaging of human eggs, zygotes, and sperm by fluorescence (confocal and epifluorescence) and electron microscopy.
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Acknowledgments
Methods described in this chapter are based on protocols developed by V. Rawe at CEGyR and during her postdoctoral training in Dr. Gerald Schatten’s lab. (Pittsburgh Development Center, Pittsburgh, PA, USA). H. Chemes contributions were developed at his laboratory in the CEDIE (National Research Council of Argentina). Special thanks to Gerald Schatten, Calvin Simerly, and Peter Sutovsky for continuous scientific support and fruitful collaborations.
Part of the information presented in this chapter is the subject of the “Summer Course in Biology of Reproduction” annually organized at CEGyR (for general information: vrawe@cegyr.com/vanerawe@hotmail.com).
Supported by CEGyR Foundation and Grants from CONICET (PIP 2565), ANPCyT (PICT 9591).
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Rawe, V.Y., Chemes, H. (2009). Exploring the Cytoskeleton During Intracytoplasmic Sperm Injection in Humans. In: Carroll, D. (eds) Microinjection. Methods in Molecular Biology, vol 518. Humana Press. https://doi.org/10.1007/978-1-59745-202-1_14
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DOI: https://doi.org/10.1007/978-1-59745-202-1_14
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