Oocyte activation and parthenogenetic development of bovine oocytes following intracytoplasmic sperm injection

Li, Xihe; Hamano, Koh‐ichi; Qian, Xiaohong; Funauchi, Katsutoshi; Furudate, Makoto; Minato, Yoshiaki

doi:10.1017/s0967199499000611

Cited by 45 publications

(27 citation statements)

References 25 publications

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“…One disadvantage of the application of exogenous activation is to increase the possibility of the parthenogenetic development [41]. In the present study, only a few blastocysts (1-2%) were obtained during 8-days culture after chemical or physical parthenogenetic treatments (Table 2).…”

Section: Discussionmentioning

confidence: 66%

A combined treatment of ionomycin with ethanol improves blastocyst development of bovine oocytes harvested from stored ovaries and microinjected with spermatozoa

et al. 2009

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Regardless of the presence of sperm-borne oocyte-activating factors, activation of bovine oocytes with exogenous activation stimuli is required for further development after intracytoplasmic sperm injection (ICSI). The present study was designed to develop a new activation regimen for improving the blastocyst yield after ICSI of bovine oocytes harvested from ovaries stored at 10-12 oC for 24 h. Following ICSI, oocytes were treated with 5 μM ionomycin for 5 min, 7% ethanol for 5 or 10 min, ionomycin followed by ethanol (5 or 10 min), ionomycin followed by 10 µg/mL Cycloheximide for 5 h, or ionomycin followed by 1.9 mM 6-dimethylaminopurine for 3 h. Across the activation regimens, the cleavage rates of ICSI oocytes (45-77%) were higher than those of parthenogenetically activated oocytes (11-21%; P<0.05). Activating the ICSI oocytes with ionomycin plus ethanol improved the blastocyst yield (29-30%) comparing to the non-treated oocytes (12%; P<0.05) but the other regimens did not (9-18%; P>0.05). The higher blastocyst yields were due to increasing the proportion of ICSI oocytes that passed through the early postfertilization events until cleavage. None of the regimens have any adverse effect on the quality of the blastocysts regarding the total cell number or the proportion of the inner cell mass cells. Thus, a new activation regimen composed from two triggers for single calcium increase has been proven effective to improve the blastocyst yield after bovine ICSI using oocytes harvested from stored ovaries.Dear Dr. John P. Kastelic, Thank you very much for prompt reply and accepting our manuscript for publication in Theriogenology. We have replied for each comment from Co-editor and the reviewer-2 in pointby-point basis.Sincerely yours, Shinichi Hochi (Corresponding author)Hany Abdalla (First author)Co-editor 1-P value has been added to the Abstract (Lines 27,29).2-The number of the references in Introduction section has been reduced. Reviewer #21-Regarding omitting the two groups (5 min ethanol group and ionomycin followed by 5 min ethanol group); We do not think that the presence of these two groups causes any difficulty to recognize the superiority of ionomycin and ethanol combination. Since activation of 7% ethanol for 5 min 4 hr after ICSI is a commonly applied method for bovine ICSI, presence of such group must be important to declare the superiority of the new combination (ionomycin and ethanol) over the methods of ethanol alone under the same experimental circumstance regarding oocyte quality, injection skill, culture condition, and so on. For the ionomycin followed by 5 min ethanol, the efficiency of this method to improve the cleavage and the blastocyst yield was similar to that of ionomycin followed by 10 min ethanol, making it possible to choose the method in which oocytes are exposed to minimum exogenous stimuli. Tables 1 and 2. 2-The time of 2 nd polar body detection has been added in footnote of 15Running head: Bovine oocyte activation after ICSI * Manuscript 2 AbstractRegardless of the pres...

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Section: Discussionmentioning

confidence: 66%

A combined treatment of ionomycin with ethanol improves blastocyst development of bovine oocytes harvested from stored ovaries and microinjected with spermatozoa

et al. 2009

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“…Presence of exogenous plasmid DNA in LI, IVF and ICSI embryos was determined by PCR, and localization of exogenous DNA into their nuclei was confirmed by FISH in ICSIderived embryos. Chemical activation following nontransgenic ICSI has been reported to improve bovine embryo development [20][21][22][23][24]. We also demonstrated that the activation in transgenic ICSI improves embryo development [25], showing different outcomes according to the chemical activation treatments [26].…”

mentioning

confidence: 67%

Efficiency of Sperm-Mediated Gene Transfer in the Ovine by Laparoscopic Insemination, In Vitro Fertilization and ICSI

Pereyra-Bonnet

Gibbons

Cueto

et al. 2011

Journal of Reproduction and Development

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Abstract. Transgenesis constitutes an important tool for pharmacological protein production and livestock improvement. We evaluated the potential of laparoscopic insemination (LI), in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) to produce egfp-expressing ovine embryos, using spermatozoa previously exposed to pCX-EGFP plasmid in two different sperm/DNA incubation treatments: "Long Incubation" (2 h at 17 C) and "Short Incubation" (5 min at 5 C). For LI, Merino sheep were superovulated and inseminated with treated fresh semen from Merino rams. The embryos were recovered by flushing the uterine horns. For IVF and ICSI, slaughterhouse oocytes were fertilized with DNA-treated frozen/thawed sperm. All recovered embryos were exposed to blue light (488 nm) to determine green fluorescent morulae and blastocysts rates. High cleavage and morulae/blastocysts rates accompanied the LI and IVF procedures, but no egfp-expressing embryos resulted. In contrast, regardless of the sperm/ plasmid incubation treatment, egfp-expressing morulae and blastocysts were always obtained by ICSI, and the highest transgenesis rate (91.6%) was achieved with Short Incubation. In addition, following the incubation of labeled plasmid DNA, after Long or Short exposure treatments, with fresh or frozen/thawed spermatozoa, only non-motile fresh spermatozoa could maintain an attached plasmid after washing procedures. No amplification product could be detected following PCR treatment of LI embryos whose zonae pellucidae (ZP) had been removed. In order to establish conditions for transgenic ICSI in the ovine, we compared three different activation treatments, and over 60% of the obtained blastocysts expressed the transgene. For ICSI embryos, FISH analysis found possible signals compatible with integration events. In conclusion, our results show that in the ovine, under the conditions studied, ICSI is the only method capable of producing exogenous gene-expressing embryos using spermatozoa as vectors. Key words: Green fluorescent protein, Sheep, Transgenesis (J. Reprod. Dev. 57: [188][189][190][191][192][193][194][195][196] 2011) ransgenesis in mammals constitutes an important tool for recombinant protein production [1,2] and also for livestock improvement [3]. Currently, the challenge is to find a high efficiency and low-cost technique for producing transgenic animals. Microinjection of foreign DNA into the male pronuclei of zygotes [4] and nuclear transfer using genetically modified somatic donor cells [5], are two commonly used methods for the generation of transgenic animals. Transgenesis by microinjection is used in mice mainly for research purposes. This procedure has also proven to be efficient in rabbits, sheep and pigs [6], but it is quite dependant on proper visualization of the male pronucleus. In addition, nuclear transfer, using genetically modified somatic donor cells is not very efficient, which can be partly explained by epigenetic reprogramming failure [7].Evidence of heterologous DNA interaction with mammalian ...

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“…In domestic species, it has been reported that failed or abnormal decondensation of sperm chromatin is frequency observed in oocytes injected with intact sperm [33,34]. Katayama et al [35] showed that oocyte activation and decondensation of sperm chromatin are delayed after injection of intact spermatozoa, which is due to the existence of the sperm plasma membrane.…”

Section: Discussionmentioning

confidence: 99%

Removal of Acrosomal Membrane from Sperm Head Improves Development of Rat Zygotes Derived from Intracytoplasmic Sperm Injection

2009

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Abstract. When intracytoplasmic sperm injection (ICSI) is applied in the rat, sperm chromatin is introduced into the oocyte together with the acrosome, which does not enter the cytoplasm of the oocyte during normal fertilization, resulting in the rat giving birth to pups. Since successful ICSI was reported in rats, but with low efficiency, it has been observed that the acrosome of the sperm head seems to have detrimental effects on the embryonic development of ICSI oocytes. To improve ICSI in rats, the effects of removal of the acrosomal membrane from rat sperm on the development of ICSI oocytes were examined. While most control (non-treated) sperm had an intact acrosomal membrane, the Triton X-100 (TX)-and lysolecithin (LL)-treated groups showed high percentages of sperm with a removed acrosomal membrane. The timing of pronuclear formation in ICSI-oocytes using TX-or LL-treated sperm was significantly accelerated compared with that of the control sperm (P<0.05). However, neither TX nor LL treatment affected amounts of PLCζ in rat sperm. The rates of offspring derived from TX-(20.3 ± 4.4%) and LL-treated sperm (19.0 ± 2.8%) were also significantly higher than that of the control group (7.6 ± 2.3%; P<0.05). Our data clearly indicate that removal of acrosomal membranes from sperm by reagents is effective for generation of offspring via ICSI in rats. Key words: Acrosomal membranes, Intracytoplasmic sperm injection, Rat (J. Reprod. Dev. 55: [475][476][477][478][479] 2009) n rats, intracytoplasmic sperm injection (ICSI) is the only way to routinely obtain offspring routinely derived from oocytes fertilized 'in vitro' using fresh and cryopreserved sperm [1], although we have previously succeeded in generation of rat offspring using cryopreserved sperm via artificial insemination ('in vivo' fertilization) [2]. Because a reliable and reproducible protocol of in vitro fertilization (IVF) using cryopreserved sperm has not been widely applied for rats, although we have established an IVF protocol very recently [3]. Successful ICSI was first reported in mice [4] and has subsequently been applied to many mammalian species [5][6][7][8][9][10]. Moreover, ICSI has been used for efficient production of not only wild-type animals but also transgenic animals [11]. In the case of rats, the protocol of ICSI has been improved by using an injection pipette with a very small outer diameter (2-4 μm), resulting in successful generation of offspring derived from the ICSI oocytes [12][13][14]. However, the efficiency is still low even though improvements such as using a small injection pipette have been applied to rat ICSI. These results imply that further improvement is required.Recently, Morozumi and Yanagimachi [15] reported that removal of both the plasma and acrosomal membranes from mouse spermatozoa before ICSI accelerates the onset of oocyte activation. They also suggested that removal of acrosomal membranes prior to ICSI is recommended for animals with large sperm acrosomes in view of the potential harmful effects of acrosomal ...

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Oocyte activation and parthenogenetic development of bovine oocytes following intracytoplasmic sperm injection

Cited by 45 publications

References 25 publications

A combined treatment of ionomycin with ethanol improves blastocyst development of bovine oocytes harvested from stored ovaries and microinjected with spermatozoa

A combined treatment of ionomycin with ethanol improves blastocyst development of bovine oocytes harvested from stored ovaries and microinjected with spermatozoa

Efficiency of Sperm-Mediated Gene Transfer in the Ovine by Laparoscopic Insemination, In Vitro Fertilization and ICSI

Removal of Acrosomal Membrane from Sperm Head Improves Development of Rat Zygotes Derived from Intracytoplasmic Sperm Injection

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