Potential of hypertonic medium treatment for embryo micromanipulation: I. Survival of rabbit embryos in vitro and in vivo following sucrose treatment

Yang, Xiangzhong; Chen, Y.; Chen, J.; Foote, R.H.

doi:10.1002/mrd.1080270205

Cited by 22 publications

(12 citation statements)

References 13 publications

(21 reference statements)

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“…It is important to point out that the sucrose-induced shrinkage of the oocytes does not significantly reduce the viability of the oocytes or the subsequent development of the embryos, as shown by our studies [10,11,24,25] and by those of others [18-23, 26, 27]. The electrolyte NaCl, on the other hand, has a deleterious effect on the viability of oocytes and is not suitable for the induction of projection for purposes of micromanipulation [31,32].…”

Section: Discussionmentioning

confidence: 52%

“…Sucrose has been widely used in embryo and oocyte cryopreservation [18][19][20][21][22][23] as well as micromanipulations, such as intracytoplasmic sperm injection and nuclear transfer [24][25][26][27]. Unable to permeate through the plasma membrane, sucrose induces water molecules to move out of the oocytes and, thus, effectively cause them to shrink.…”

Section: Discussionmentioning

confidence: 99%

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Hypertonicity-Induced Projections Reflect Cell Polarity in Mouse Metaphase II Oocytes: Involvement of Microtubules, Microfilaments, and Chromosomes1

Liu

Sung

Tian

et al. 2002

Biology of Reproduction

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A previous study showed that with hypertonic sucrose treatment, a projection is formed in mouse metaphase II (MII) oocytes in proximity to the spindle and chromosomes, where a polarized cortical domain is located. However, little is known about the mechanisms involved in this process. Here, we designed a series of experiments to test the hypothesis that hypertonicity is the induction factor for the formation of projections in mouse MII oocytes. Our hypothesis was supported by the following evidence: 1) different concentrations of sucrose affected the formation and shape of projections, whereas serum or basic media had little effect; 2) other hypertonic sugar solutions could also induce projection formation; and 3) projections could also be induced by hypertonic NaCl solution. We then tested the hypothesis that the cytoskeleton was involved in the formation of hypertonicity-induced projections. This was investigated by culturing MII- and germinal vesicle-stage mouse oocytes in the presence or absence of cytoskeletal inhibitors, including cytochalasin B (disruption of actin filaments), nocodazole (disruption of microtubules), and taxol (polymerization of tubulin molecules). We found that none of the cytoskeletal inhibitors alone could prevent hypertonicity-induced projection formation, whereas the combination of cytochalasin B with nocodazole or with taxol blocked the formation of these projections in most matured oocytes. When immature oocytes were incubated in cytochalasin B, but not in nocodazole or taxol, the formation of an actin-rich domain and the peripheral positioning of the spindle were blocked during maturation; hence, no projections were formed, even after hypertonic sucrose treatment. Based on these observations, we propose that three components are necessary for projection formation: 1) a polarized cortical patch (e.g., an actin-rich domain), 2) rigid submembrane structures (e.g., a spindle and/or chromosomes), and 3) solid connections between the above. Any disturbance of one of these factors will affect the hypertonicity-induced projection formation. Hypertonicity-induced projection in mouse oocytes thus provides an experimental model for studies regarding cell polarity and the interaction between membrane and submembrane components.

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Section: Discussionmentioning

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Hypertonicity-Induced Projections Reflect Cell Polarity in Mouse Metaphase II Oocytes: Involvement of Microtubules, Microfilaments, and Chromosomes1

Liu

Sung

Tian

et al. 2002

Biology of Reproduction

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“…Embryos of both species examined in the present study were considered normal embryos on the basis of the embryonic development data in our previous publications [14,16,17]. The fact that more bovine blastomeres were classified as "uncertain" suggests that cattle embryos produced by IVM/IVF procedures were of lower quality.…”

Section: General Commentsmentioning

confidence: 98%

“…Sexually mature Dutch-belted rabbits were superovulated [16]. Eight-to sixteen-cell stage embryos were collected surgically from the oviducts of superovulated does 46 h after insemination and injection with 0.5 mg of pituitary LH (PLH; Burns-Biotech, Inc., Omaha, NE).…”

Section: Collection Of Rabbit Embryosmentioning

confidence: 99%

“…The zona pellucida of the bovine embryos, and the mucin coat plus the zona pellucida of the rabbit embryos, were removed by incubation in prewarmed acidic DPBS-PVA (pH = 2.5) for approximately 2 min at room temperature and then incubated with 0.5% pronase in DPBS-PVA at 39°C for 5-15 min [16]. The zona-free embryos in both cattle and rabbits were then incubated in Ca 2 +-free DPBS-PVA at 39°C for 20 min.…”

Section: Removal Of Accessory Coats and Isolation Of Single Embryo Blmentioning

confidence: 99%

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Analysis of Polarity of Bovine and Rabbit Embryos by Scanning Electron Microscopy1

Koyama

Suzuki

Yang

et al. 1994

Biology of Reproduction

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Cellular polarization during preimplantation development of the embryo is believed to be a crucial event in the transition of a zygote to a blastocyst stage embryo with morphologically and functionally differentiated cell types. While extensive studies have been done on polarity development in mouse embryos, little information is available in other species, particularly in cattle. The objective of this study was to examine the initiation of polarity by microvilli distribution on blastomeres of cattle and rabbit embryos using scanning electron microscopy (SEM). Bovine embryos were obtained by in vitro fertilization of in vitro-matured follicular oocytes. Rabbit embryos of various stages were collected from superovulated rabbits. Intact embryos and isolated blastomeres were examined in both species. Blastomeres from 1- to 8-cell embryos in both cattle and rabbits showed no polarity. The onset of transitional polarization of microvillous distribution occurred in some blastomeres of cattle embryos at the 9- to 15-cell stage; but typical, distinct polarity was not manifested until after the 16-cell stage with approximately 40% polar cells per embryo. In the rabbit blastomere polarity occurred one cell cycle later, with 46% polar cells per embryo after the 32-cell stage. The difference in cell numbers at the time polarity is evident is probably related to the different cell stages for embryo compaction and blastocyst formation in the two species.

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Improved developmental potential of rabbit oocytes fertilized by sperm microinjection into the perivitelline space enlarged by hypertonic media

1990

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The objectives of the present study were: 1) to develop a simple and more efficient technique for sperm microinjection than is currently available, using the rabbit as a model, and 2) to evaluate the development of rabbit oocytes fertilized by single or multiple sperm microinjection. Hyperosmotic sucrose in phosphate-buffered saline (SPBS) was employed to dehydrate oocytes to increase the perivitelline space for sperm microinjection and prevent possible injury to the vitellus. In the first experiment, 58% (n = 29) oocytes treated with 0.5 M SPBS developed to morulae following multiple sperm microinjection compared, respectively, to 47% (n = 34) and 60% (n = 15) for control IVF with or without sucrose exposure (P greater than 0.05). Blastocyst development from microinjected oocytes, however, was much lower (P less than 0.05) than that of controls (14% vs. 42% and 40%, respectively). Sham operation by puncturing the zona pellucida of the sucrose-treated oocytes with the microinjection pipette did not increase parthenogenesis (P greater than 0.05). In Experiment 2 a smaller-size injection pipette and shorter sucrose exposure time after sperm microinjection resulted in 41% (n = 42) of the oocytes developing into blastocysts for the microinjection group, whereas only 21% (n = 24) developed to blastocysts in the control IVF group (P less than 0.05). When relatively older oocytes (17 hr post ovulation injection) were used to test if microinjection could reduce the time to fertilization and cleavage (Expt. 3), an average of 27% (n = 63) blastocysts resulted from microinjection vs. 0% (n = 28) for the control IVF group.

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Potential of hypertonic medium treatment for embryo micromanipulation: I. Survival of rabbit embryos in vitro and in vivo following sucrose treatment

Cited by 22 publications

References 13 publications

Hypertonicity-Induced Projections Reflect Cell Polarity in Mouse Metaphase II Oocytes: Involvement of Microtubules, Microfilaments, and Chromosomes1

Hypertonicity-Induced Projections Reflect Cell Polarity in Mouse Metaphase II Oocytes: Involvement of Microtubules, Microfilaments, and Chromosomes1

Analysis of Polarity of Bovine and Rabbit Embryos by Scanning Electron Microscopy1

Improved developmental potential of rabbit oocytes fertilized by sperm microinjection into the perivitelline space enlarged by hypertonic media

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