Development of porcine embryos and offspring after intracytoplasmic sperm injection with liposome transfected or non-transfected sperm into <i>in vitro</i> matured oocytes

Lai, Liangxue; Sun, Qing‐Yuan; Wu, Guangming; Murphy, Clifton N.; Kühholzer, B.; Park, Kwang-Wook; Bonk, Aaron J.; Day, Bill; Prather, Randall S.

doi:10.1017/s0967199401001393

Cited by 77 publications

(54 citation statements)

References 20 publications

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“…6I). Porcine embryos are produced routinely by both IVF and SCNT methods described here and are capable of implantation and development to term after embryo transfer (Macháty et al 1998, Lai et al 2001, Lai & Prather 2003. If allowed to develop to the blastocyst stage in vitro, they have a low blastomere count and poor morphology not comparable to the in vivo-generated blastocysts (see Fig.…”

Section: Resultsmentioning

confidence: 99%

Expression and proteasomal degradation of the major vault protein (MVP) in mammalian oocytes and zygotes

Šutovský¹,

Manandhar²,

Laurinčík³

et al. 2005

Reproduction

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Major vault protein (MVP), also called lung resistance-related protein is a ribonucleoprotein comprising a major part (>70%) of the vault particle. The function of vault particle is not known, although it appears to be involved in multi-drug resistance and cellular signaling. Here we show that MVP is expressed in mammalian, porcine, and human ova and in the porcine preimplantation embryo. MVP was identified by matrix-assisted laser-desorption ionization-time-of-flight (MALDI-TOF) peptide sequencing and Western blotting as a protein accumulating in porcine zygotes cultured in the presence of specific proteasomal inhibitor MG132. MVP also accumulated in poor-quality human oocytes donated by infertile couples and porcine embryos that failed to develop normally after in vitro fertilization or somatic cell nuclear transfer. Normal porcine oocytes and embryos at various stages of preimplantation development showed mostly cytoplasmic labeling, with increased accumulation of vault particles around large cytoplasmic lipid inclusions and membrane vesicles. Occasionally, MVP was associated with the nuclear envelope and nucleolus precursor bodies. Nucleotide sequences with a high degree of homology to human MVP gene sequence were identified in porcine oocyte and endometrial cell cDNA libraries. We interpret these data as the evidence for the expression and ubiquitin-proteasome-dependent turnover of MVP in the mammalian ovum. Similar to carcinoma cells, MVP could fulfill a cell-protecting function during early embryonic development.

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Section: Resultsmentioning

confidence: 99%

Expression and proteasomal degradation of the major vault protein (MVP) in mammalian oocytes and zygotes

Šutovský¹,

Manandhar²,

Laurinčík³

et al. 2005

Reproduction

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“…This route circumvents membrane fusion and therefore does not rely on sperm internalising exogenous DNA. It results in conclusive DNA uptake and reproducible reporter gene expression in transgenic embryos and animals (Perry et al 1999, Chan et al 2000, Lai et al 2001, Pereyra-Bonnet et al 2008, Garcia-Vazquez et al 2009, Wu et al 2009, Shadanloo et al 2010.…”

Section: Introductionmentioning

confidence: 99%

Exposure to DNA is insufficient for in vitro transgenesis of live bovine sperm and embryos

Eghbalsaied¹,

Ghaedi²,

Laible³

et al. 2013

Reproduction

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Transgenic mammals have been produced using sperm as vectors for exogenous DNA (sperm-mediated gene transfer (SMGT)) in combination with artificial insemination. Our study evaluated whether SMGT could also be achieved in combination with IVF to efficiently produce transgenic bovine embryos. We assessed binding and uptake of fluorescently labelled plasmids into sperm in the presence of different concentrations of dimethyl sulphoxide or lipofectamine. Live motile sperm displayed a characteristic punctuate fluorescence pattern across their entire surface, while uniform postacrosomal fluorescence was only apparent in dead sperm. Association with sperm or lipofection reagent protected exogenous DNA from DNase I digestion. Following IVF, presence and expression of episomal and non-episomal green fluorescent protein (GFP)-reporter plasmids was monitored in oocytes and embryos. We found no evidence of intracellular plasmid uptake and none of the resulting zygotes (nZ96) and blastocysts were GFP positive by fluorescence microscopy or genomic PCR (nZ751). When individual zona-free oocytes were matured, fertilised and continuously cultured in the presence of episomal reporter plasmids until the blastocyst stage, most embryos (38/68Z56%) were associated with the exogenous DNA. Using anti-GFP immunocytochemistry (nZ48) or GFP fluorescence (nZ94), no GFP expression was detected in blastocysts. By contrast, ICSI resulted in 18% of embryos expressing the GFP reporter. In summary, exposure to DNA was an inefficient technique to produce transgenic bovine sperm or blastocysts in vitro.

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“…In case of using in vitro matured pig oocytes for ICSI, we think that the artificial treatment for induction of oocyte activation (resumption of meiosis, estrusion of a 2nd polar body, pronuclear formation and DNA replication) is one of the most important factors for fertilization and embryonic development. Because electrical stimulation promoted pronuclear formation and development to the blastocyst stage [30][31][32]. These positive effects did not depend on status of sperm membrane and nuclei [27] (Table 1).…”

Section: Responsibility Of Oocyte Activation For Pronuclear Formationmentioning

confidence: 87%

Factors Affecting Fertilization and Embryonic Development During Intracytoplasmic Sperm Injection in Pigs

Nakai

Kashiwazaki

Ito

et al. 2011

Journal of Reproduction and Development

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Abstract. In intracytoplasmic sperm injection (ICSI) technique, a sperm was injected into ooplasm directly using a glass pipette. The fertilization physiology in ICSI is considered quite different from that of the natural fertilization. The different mechanisms for fertilization may be the causes of various results in ICSI. In this paper, we focus on the state of sperm membranes, nuclear or DNA integrity during ICSI procedure and discuss the influence of these factors on fertilization and embryonic development. We also introduce some examples in application of ICSI for new technologies in pigs. Key words: Embryonic development, Fertilization, ICSI, Pig (J. Reprod. Dev. 57: [183][184][185][186][187] 2011) uring natural fertilization, sperm needs several steps; e.g., attachment to zona pellucida, penetration through zona pellucida with releasing of acrosomal enzymes from acrosomal caps, fusion of sperm membrane with oolemma, decondensation and recondensation of sperm nucleus after incorporation into ooplasm, and finally formation of male pronucleus [1]. Therefore, if sperm have no or faint motility, such sperm cannot reach and penetrate an oocyte resulting in a failure of fertilization. On the other hand, by intracytoplasmic sperm injection (ICSI), a whole spermatozoon or a sperm nucleus can be deposited directly into ooplasm by a glass pipette. Therefore, ICSI technique enables production of fertilized oocytes even if spermatozoa lack their competence for physiological fertilization.As mentioned above, spermatozoa can be brought into ooplasm by ICSI without any physiological events. In other words, spermatozoa bring intact membrane and acrosomal enzymes into ooplasm. Furthermore, there causes a possibility that sperm with some abnormalities may participate in fertilization and further embryonic development resulting in offspring, because spermatozoa are injected into ooplasm by artificial procedure without physiological selection processes. In this paper, we discuss about sperm factors which may affect the completion of fertilization by ICSI in pigs as a model for large domestic animals. Furthermore, recently the combination of ICSI and some other techniques has been expected as novel procedure for conserving male genetic resource. For completion of this technology, ICSI is the most essential procedure. We introduced the recent results in our laboratory for these new approaches. Carrying of Acrosomal Membrane and Their Enzymes Into OoplasmAt the first step of fertilization, sperm have to travel through the cumulus cells and their matrix and then penetrate the zona pellucida. In natural fertilization, sperm release acrosomal enzymes from acrosome cap (acrosome reaction), move forward by a driving force of sperm tail. Therefore, after the penetration of zona pellucida, sperm without outer-acrosomal membrane and acrosomal enzymes reach to the oolemma. However, by ICSI, sperm do not lose those constructions and those with their complete form are injected into ooplasm. Therefore, it is important to discuss the effe...

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Development of porcine embryos and offspring after intracytoplasmic sperm injection with liposome transfected or non-transfected sperm into in vitro matured oocytes

Cited by 77 publications

References 20 publications

Expression and proteasomal degradation of the major vault protein (MVP) in mammalian oocytes and zygotes

Expression and proteasomal degradation of the major vault protein (MVP) in mammalian oocytes and zygotes

Exposure to DNA is insufficient for in vitro transgenesis of live bovine sperm and embryos

Factors Affecting Fertilization and Embryonic Development During Intracytoplasmic Sperm Injection in Pigs

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