Fertilization by sperm injection in cattle

Goto, Kazufumi; Kinoshita, Akihiro; Takuma, Yoshihiro; Ogawa, Kohei

doi:10.1016/0093-691x(90)90662-d

Cited by 23 publications

(17 citation statements)

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“…SI-0 is the most direct microinsemination method; however, newborns were obtained only from rabbits [5] and bovines [3], and thus it has not been clinically applied. Nevertheless, SI-0 is excellent as a study measure for clarifying the mechanism of fertilization or examining the gamete reproductive ability, and is used by many researchers.…”

Section: Discussionmentioning

confidence: 99%

Microinsemination of Rabbit Oocytes with Heat-Treated Sperm: Embryonic Development

Hoshi

Yazawa

Sato

1992

Archives of Andrology

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A study was conducted to evaluate the ability of rabbit oocyte to develop to the pronuclear stage and subsequent development if injected with heat-treated sperm. The rate of fertilization of eggs after microinjection of cauda epididymal spermatozoa heated at 60°C for 30 min was 36%; the rate of cleavage was 24 %. The rate of fertilization after microinjection of heat-treated sperm nuclei was 43 % ; the rate of cleavage was 26%. On the other hand, the rate of fertilization of eggs injected with sperm without heat treatment was 43 %, the rate of cleavage was 29 %, and there was no difference between eggs microinjected with heat-treated spermatozoa or sperm nuclei and those not heat treated. Of the viable eggs injected with heat-treated sperm, 16% developed to the 6-to 8-cell stage 72 h after injection.

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Section: Discussionmentioning

confidence: 99%

Microinsemination of Rabbit Oocytes with Heat-Treated Sperm: Embryonic Development

Hoshi

Yazawa

Sato

1992

Archives of Andrology

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“…A recent advanced assisted reproductive technique, intracytoplasmic sperm injection (ICSI), has demonstrated that killed sperm or only sperm heads separated from tails were still useful for making normal offspring [2,10]. The fact that the sperm nucleus is capable of fertilization regardless of whether it is dead or alive gives us an exciting suggestion for a strategy to rescue endangered or already extinct animals.…”

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confidence: 99%

“…This feature is due to the dramatic restructuring of chromatin during spermiogenesis, where some spermatogenic cell specific nuclear proteins, i.e., protamine and transition protein, are expressed instead of the histones [1,3,11]. The whole DNA of sperm is, therefore, believed to be very resistant to various environmental, physical and chemical insults, for example, exposure to high temperature and freezing-thawing [9,12,19].A recent advanced assisted reproductive technique, intracytoplasmic sperm injection (ICSI), has demonstrated that killed sperm or only sperm heads separated from tails were still useful for making normal offspring [2,10]. The fact that the sperm nucleus is capable of fertilization regardless of whether it is dead or alive gives us an exciting suggestion for a strategy to rescue endangered or already extinct animals.…”

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confidence: 99%

Comparison of the Nuclear DNA Stability against Freezing-Thawing and High Temperature Treatments between Spermatozoa and Somatic Cells.

Ohsako

Nagano

Sugimoto

et al. 1997

The Journal of Veterinary Medical Science

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ABSTRACT. Thermostability of sperm genome against freezing-thawing and high temperature treatments was assessed by comparing the degradation patterns of genomic DNAs from epididymal sperms and somatic tissues. Golden hamster liver, kidney, epididymal sperm, and testis were frozen and thawed repeatedly, or incubated in a hot water bath. Genomic DNAs were isolated and then separated by agarose gel electrophoresis. It was revealed that the size of sperm genomic DNA was hardly changed after freezing-thawing treatment, however, the DNA sizes of the other three tissues were gradually reduced with an increasing number of freezing-thawing cycles. In contrast, high temperature treatment appears to damage not only the genomic DNAs of somatic cells but also those of spermatozoa. -KEY WORDS: DNA, sperm, temperature.J. Vet. Med. Sci. 59(11): 1085-1088, 1997 effects of freezing-thawing treatments, they were all put in sterile polypropylene tubes after washing with phosphate buffered saline (PBS), and divided into four groups. One group was kept in a freezer at 20°C quickly, and the other three groups were frozen in liquid nitrogen, and then thawed by 10 min incubation in a shaking water bath at 27°C. This freezing-thawing cycle was repeated 5, 10, or 20 times and each sample was then kept in a freezer at 20°C until genomic DNA isolation. Sperms were collected by partially cutting caudal epididymis in TBS solution (10 mM Tris-HCl, 150 mM NaCl, pH 7.4). An approximate 0.1 ml volume of sperm pellet and all the other tissue homogenates were resuspended with 1.4 ml lysis buffer containing 150 mM NaCl, 15 mM Sodium acetate, 1 mM EDTA, 1% SDS, 0.01 mg/ml Proteinase K (Nacalai Tesque, Inc., Tokyo, Japan), and then incubated in a shaking water bath at 50°C for 1 hr. In order to remove nuclear basic proteins, 167 µl of 1 M 2-mercaptoethanol (Nacalai Tesque, Inc. Tokyo, Japan) was added followed by 3 hr incubation. After a two-fold phenol extraction, they were digested with 100 µg/ml RNase A (Sigma, MO, U.S.A.) at 37°C for 1 hr. After phenol/chloroform extraction and ethanol precipitation, DNAs were resuspended in TE (10 mM Tris, 1 mM EDTA, pH 8.2), and their concentration was then measured by UV spectrophotometer. All the isolated DNA samples were evenly separated by 0.8% agarose gel electrophoresis and observed with a UV transilluminator. With the same protocol, the same tissues were frozen at 80°C in a deep freezer or at 20°C in a freezer for 1 hr and thawed at 27°C in a shaking water bath. The cycle was repeated 5, 10, or 20 times and the samples were then kept at 20°C in a freezer until DNA isolation. Additionally, to examine effects of high temperature treatment, one set of tissues was incubated at 50°C in a hot shaking water bath for 30 min, 1 hr, 3 hr, and 6 hr, respectively, and then kept at 20°C in a freezer until DNA isolation.In the experiments of freezing-thawing treatment, the core DNA sizes of isolated genomic DNAs from liver, kidney and testis on the gels, approximately 100 kb, were reduced Mammalian sperm DNA is the mos...

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“…Many childless couples who are unable to conceive by conventional in vitro fertilization and other treatments now have a fairly good chance to have their own children by ICSI. Animals in which ICSI has been successfully used to produce live offspring include mice (2), golden hamsters (3), rats (4), rabbits (5), cattle (6), sheep (7), horses (8), cats (9), pigs (10), and monkeys (11).…”

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Incorporation of the acrosome into the oocyte during intracytoplasmic sperm injection could be potentially hazardous to embryo development

Morozumi

Yanagimachi

2005

Proc. Natl. Acad. Sci. U.S.A.

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In mice and humans, a normal offspring can be obtained by injecting a single spermatozoon into an oocyte, the process called intracytoplasmic sperm injection (ICSI). When three or more mouse spermatozoa with intact acrosomes were injected into individual mouse oocytes, an increasing proportion of oocytes became deformed and lysed. Oocytes did not deform and lyse when acrosome-less spermatozoa were injected, regardless of the number of spermatozoa injected. Injection of more than four human spermatozoa into a mouse oocyte produced vacuole-like structures in each oocyte. This vacuolation did not happen when spermatozoa were freed from acrosomes before injection. Hamsters, cattle, and pigs have much larger acrosomes than the mouse or human. Injection of a single acrosome-intact hamster, bovine, and porcine spermatozoon deformed and lysed many or all mouse oocytes. This deformation did not happen when these spermatozoa were freed from acrosomes before ICSI, regardless of the number of spermatozoa injected. Because trypsin and hyaluronidase mimicked the action of acrosome-intact spermatozoa, it is likely that the acrosomal enzymes deform and lyse the oocytes. Injection of small amounts of trypsin and hyaluronidase into normally fertilized mouse eggs disturbed their pre-and postimplantation development. In view of potentially harmful effects of acrosomal enzymes on embryo development, the removal of acrosomes before ICSI is recommended for animals with large sperm acrosomes. The removal of acrosomes may increase the efficiency of ICSI in these animals. Although human and mouse spermatozoa do not need to be freed from acrosomes, the removal of acrosomes before ICSI is theoretically preferable. assisted fertilization ͉ bovine ͉ human ͉ mouse ͉ porcine

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Fertilization by sperm injection in cattle

Cited by 23 publications

References 0 publications

Microinsemination of Rabbit Oocytes with Heat-Treated Sperm: Embryonic Development

Microinsemination of Rabbit Oocytes with Heat-Treated Sperm: Embryonic Development

Comparison of the Nuclear DNA Stability against Freezing-Thawing and High Temperature Treatments between Spermatozoa and Somatic Cells.

Incorporation of the acrosome into the oocyte during intracytoplasmic sperm injection could be potentially hazardous to embryo development

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