Effects of Sperm Pretreatment on Efficiency of ICSI-Mediated Gene Transfer in Pigs
| Autor: | S. Ueno, Ryo Tomii, H. Saito, K. Hiruma, Mayuko Kurome, Hiroshi Nagashima |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Male
medicine.medical_treatment Green Fluorescent Proteins Sus scrofa Acrosome reaction Intracytoplasmic sperm injection Cryopreservation Animals Genetically Modified Embryo Culture Techniques Andrology Sonication Pregnancy medicine Animals Sperm Injections Intracytoplasmic Transgenes Blastocyst reproductive and urinary physiology urogenital system Chemistry Acrosome Reaction Embryogenesis Gene Transfer Techniques Embryo Sperm medicine.anatomical_structure Sperm Tail embryonic structures Oocytes Sperm Head Female Animal Science and Zoology Exogenous DNA |
| Zdroj: | Journal of Reproduction and Development. 53:1217-1226 |
| ISSN: | 1348-4400 0916-8818 |
| DOI: | 10.1262/jrd.19069 |
| Popis: | Intracytoplasmic sperm injection (ICSI)-mediated gene transfer has recently been shown to be an effective technique for producing transgenic pigs; however, the types of sperm pretreatment having the most beneficial effects on post-ICSI embryogenesis or transgenic efficiency have not been clarified. In the present study, we performed ICSI-mediated gene transfer using pig sperm subjected to various pretreatments and determined the developmental potential of sperm-injected oocytes and introduction efficiency of exogenous DNA. Embryos were then transferred to recipient pigs to confirm gene transfer efficiency during the fetal period. When ICSI was performed using unfrozen sperm heads with tails removed by piezo-pulse, the rates of blastocyst formation (14.2%, 17/120) and transgene (EGFP) expression (11.8%, 2/17) were both low. When unfrozen sperm heads were used that were removed by sonication, EGFP expression efficiency (11/21, 52.4%) improved significantly (P |
| Databáze: | OpenAIRE |
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