Development of Parthenogenetic Rat Embryos1

Krivokharchenko, Alexander; Popova, Elena; Zaitseva, Ioulia; Vilianovich, Larissa; Ganten, Detlev; Bäder, Michael

doi:10.1095/biolreprod.102.006494

Cited by 45 publications

(40 citation statements)

References 48 publications

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“…Second polar body extrusion and fragmented cytoplasm increased as CM exposure exceeded the optimum time. A small fraction (6%) of mouse MII oocytes cultured in DMEM-HG or CZB without any known parthenogenetic stimulants initiated parthenogenetic activation spontaneously, which is a common phenomenon in lower vertebrate species and amphibians (28).…”

Section: Discussionmentioning

confidence: 99%

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Feng

Zhou

Ling

et al. 2009

Braz J Med Biol Res

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Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca 2+ ] i ), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca 2+ ] i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 μg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca 2+ ] i oscillations, completion of meiosis and parthenogenetic development.

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Section: Discussionmentioning

confidence: 99%

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Feng

Zhou

Ling

et al. 2009

Braz J Med Biol Res

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“…In fact, it is well documented in several mammalian species that complete activation and pronuclear formation is dependent upon the age of the oocyte with regard to ovulation. In general, while aged oocytes are easily activated even with artificial stimuli inducing a single Ca 2þ rise, recently ovulated oocytes often arrest at MIII and repetitive Ca 2þ transients or combined treatments with protein synthesis or phosphorylation inhibitors are required for the completion of meiosis and entry into interphase in these young oocytes (Collas et al 1989, Kubiak et al 1993, Presicce & Yang 1994, Swann & Ozil 1994, Krivokharchenko et al 2003. Although it is beyond the scope of the present study, further analysis of oocyte maturation kinetics after hCG injection, quantification of IP 3 R levels and measurement of the Ca 2þ transients induced by ethanol and strontium in oocytes from the four strains of mice could be performed to assess this possibility.…”

Section: Discussionmentioning

confidence: 99%

Effect of genetic background and activating stimulus on the timing of meiotic cell cycle progression in parthenogenetically activated mouse oocytes

Ibáñez¹,

Albertini²,

Overström³

2005

Reproduction

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With the aim of investigating the effects of oocyte genotype and activating stimulus on the timing of nuclear events after activation, oocytes collected from hybrid B6D2F1, inbred C57BL/6 and outbred CF-1 and immunodeficient nude (NU/1 ) females were activated using ethanol or strontium and fixed at various time-points. Meiotic status, spindle rotation and second polar body (PB2) extrusion were monitored by fluorescence microscopy using DNA-, microtubule-and microfilament-selective probes. Although activation efficiency was similar in all groups of oocytes, a significant percentage of CF-1 and NU/1 oocytes treated with ethanol and of C57BL/6 oocytes treated either with ethanol or strontium failed to complete activation and became arrested at a new metaphase stage (MIII) after PB2 extrusion. C57BL/6 oocytes also showed slower release from MII arrest but faster progression to telophase (TII) after ethanol exposure, and they exhibited the most rapid exit from TII under both activation treatments. Strontium caused delayed meiotic resumption, spindle rotation and PB2 extrusion, but rapid TII exit, in B6D2F1, CF-1 and NU/1 oocytes when compared with ethanol. Compared with all other strains, NU/1 oocytes were significantly slower in completing spindle rotation and PB2 extrusion, irrespective of the activating stimulus, and a significant decrease in activation rates and pace of meiotic progression was observed after strontium exposure. Thus, our findings demonstrated that the kinetics of meiosis resumption and completion, spindle rotation and PB2 extrusion following parthenogenetic activation depends on both genotype-specific factors and on the activation treatment applied.

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“…Information on the effect of oocyte age on the response to activation treatment in the rat is very limited. A study in Wistar rats [8] showed that the use of ethanol for parthenogenetic activation was highly effective only for aged oocytes (22-2 4 h after hCG). However, the efficiencies of activation and cleavage rate were equal for all ages of oocytes when they were treated with strontium [8].…”

Section: Discussionmentioning

confidence: 99%

“…Since activation of the recipient oocyte is a crucial step in successful nuc lear transfer techniques [8 ], inco mplete activation of reconstructed embryos may in part be responsible for the difficulty of rat nuclear transfer. At present, the available information regarding parthenogenetic activation of rat oocytes is limited.…”

mentioning

confidence: 99%

“…A more recent study showed that Wistar rat oocytes could be activated by electrical stimulation, ethanol or strontium treatment. However, the efficiency of blastocyst formation was low (12.1-29.6%) and dramatic variations existed among individual rats in the efficiency of parthenogenetic activation of oocytes and developmental ability of the resulting embryos [8]. These results suggested that research into parthenogenetic activation of rat oocytes was insufficient, and further investigation was needed.…”

mentioning

confidence: 99%

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Determination of Optimal Conditions for Parthenogenetic Activation and Subsequent Development of Rat Oocytes In Vitro

Mizutani

Jiang

Mizuno

et al. 2004

Journal of Reproduction and Development

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Abstract. The present study was undertaken to determine optimal conditions for parthenogenetic activation and subsequent development of rat oocytes. Oocytes from immature Wistar-Imamichi (WI) and Sprague Dawley (SD) rats were activated by electrical stimulation in combination with 6-dimethylaminopurine (6-DMAP) to assess whether different rat strains display different responses to activation treatment. Since the cleavage rates of activated oocytes were significantly higher in WI than SD strain rats, WI rats were used for the subsequent experiments to determine the effects of post-hCG time, culture duration, different activation protocols (electrical stimulation with 6-DMAP or ionomycin with 6-DMAP) and osmolarity of the activation medium on the activation and subsequent development of WI rat oocytes. For oocytes activated by electrical stimulation combined with 6-DMAP, the percentages of oocytes that were activated and that developed to blastocysts were higher when oocytes were collected at 18-20 h than at any other time points after hCG injection (16, 22-24 h). Culturing for 2-6 h before activation treatment markedly decreased the percentage of activated oocytes that developed to beyond the four-cell stage. There were no differences in the percentages of oocytes with pronuclear formation and subsequent development to the two-cell and blastocyst stages between oocytes that were activated by electrical stimulation or ionomycin, both followed by 6-DMAP treatment. Activation of oocytes by ionomycin and 6-DMAP, both in low osmolarity media (246 mOsM), markedly increased the cleavage rates and percentages of high quality blastocysts (71%). The optimal conditions determined in the present study with simplified activation protocols and high efficiency of activation and subsequent development of WI rat oocytes will be helpful for further research involving nuclear transfer in the rat.

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Development of Parthenogenetic Rat Embryos1

Cited by 45 publications

References 48 publications

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Effect of genetic background and activating stimulus on the timing of meiotic cell cycle progression in parthenogenetically activated mouse oocytes

Determination of Optimal Conditions for Parthenogenetic Activation and Subsequent Development of Rat Oocytes In Vitro

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