Plasma membrane and cytocortex alterations in frozen/thawed mouse embryos deprived of the zona pellucida

Martí, M.; Grossmann, Mark; Santaló, Josep; Egozcue, J.; Ponsà, M.

doi:10.1002/(sici)1097-010x(19980101)280:1<38::aid-jez5>3.0.co;2-i

Cited by 2 publications

(1 citation statement)

References 41 publications

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“…Detrimental impact of ROS [3][4][5], and cryopreservation [6][7][8] on preimplantation embryos is well documented. On the other hand, relatively poor and preliminary information regarding the influence of Fas/FasL system on preimplantation embryo development exists [9,10].…”

Section: Introductionmentioning

confidence: 55%

Protective potential of SCF for mice preimplantation embryos cultured in vitro in suboptimal conditions

Głąbowski

Wiszniewska

Kurzawa

2008

J Assist Reprod Genet

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Purpose To examine the effect of stem cell factor (SCF) to embryos exposed to detrimental factors. Methods Mice embryos cultured in control medium or Exp.1. with FasL or FasL+SCF Exp.2. with hydrogen peroxide (HP) or HP+SCF; Exp.3. frozen-thawed and cultured with or without SCF. Immunohistochemistry for Fas and c-kit receptors was performed in blastocysts. Blastocyst rates, total numbers of blastocyst cells (TB) and inner cell mass cell counts (ICM) were determined. Results Immunohistochemical studies revealed expression of both Fas and c-kit in blastocyst cells. Exp.1. Significantly more blastocysts were found in control when compared to FasL group and to FasL+SCF group. TB and ICM counts in control and FasL+SCF group were significantly higher comparing to FasL group. Exp.2. We found significant differences between three groups in all three evaluated parameters. The highest blastocyst rates, TB and ICM counts were found in control, lower in HP+SCF group and the worst in HP group. Exp.3. No significant differences in TB and ICM counts were found. More embryos formed blastocyst in control than in two cryopreserved groups. Blastocyst rates did not differ between two cryopreserved groups. Conclusion SCF may improve culture of embryos exposed to unfavorable milieu.

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Section: Introductionmentioning

confidence: 55%

Protective potential of SCF for mice preimplantation embryos cultured in vitro in suboptimal conditions

Głąbowski

Wiszniewska

Kurzawa

2008

J Assist Reprod Genet

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Characteristics of the cell membrane fluidity, actin fibers, and mitochondrial dysfunctions of frozen‐thawed two‐cell mouse embryos

Ahn

Sohn

Kwon

et al. 2002

Molecular Reproduction Devel

101

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Physical and chemical alterations caused by the freezing and thawing and their effects on survivals/developments in vitro were investigated. Of a total of 452 two-cell mouse embryos, the overall survival rate of the frozen-thawed embryos was 76.1% (344/452). The blastocyst formation of the frozen-thawed embryos was 32.6% (44/136) compared to 74.5% (117/157) in the fresh embryos (P<0.05). The total number of cells in a blastocyst also decreased from 96.0 +/- 19.0 (n=26) in the fresh embryos to 42.0 +/- 11 .34 (n=30) in the frozen-thawed embryos (P<0.05). Fluorescence recovery after photobleaching (FRAP) measurement revealed about 5-fold decrease in the cell membrane fluidity with a characteristic time constant (tau) of 1.46 +/- 0.13 sec (n=5) in the frozen-thawed embryos as opposed to 0.28 +/- 0.04 sec (n=5) in the fresh embryos (P<0.05). The relative amount of H(2)O(2) in an embryo as quantified by the fluorescence intensity of 2',7'-dichlorofluorescein (DCF) showed 62.8 +/- 23.5 (n=24) and 34.2 +/- 14.5 (n=20) in the frozen-thawed embryos and in the fresh embryos, respectively (P<0.05). The distribution of actin filaments in the frozen-thawed embryos revealed an uneven distribution, particularly discontinuities at the "actin band," which contrasted to an even distribution shown in the fresh embryos. Mitochondrial staining by Rhodamine 123 showed that there was no significant difference between the two treatments in the number and in the distribution of viable mitochondria, but a marked aggregation was seen in the arrested embryos. No Annexin V binding was detected in two-cell or four-cell embryos while the binding was positive in the arrested embryos. The mitochondrial membrane potential measured by a membrane potential-sensitive fluorescent probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazol- carbocyanine iodide (JC-1) revealed a marked depolarization in the frozen-thawed embryos. Finally, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end-labeling (TUNEL) was employed to quantify the DNA fragmentation. In 75.0% cells of blastocysts (n=24) in the frozen-thawed embryos, the DNA fragmentation was detected as opposed to 37.0% in the fresh embryos (n=20) (P<0.05). Taken together, it is proposed that during the cryopreservation, two-cell mouse embryos are subjected to physical and chemical alterations, including destruction of the cell membrane integrity, redistribution of actin fibers, mitochondrial depolarizations, and increased reactive oxygen species (ROS) productions, which then may trigger the apoptotic cascade leading to a decrease in the survival rate and in the developmental rate of the embryos.

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Plasma membrane and cytocortex alterations in frozen/thawed mouse embryos deprived of the zona pellucida

Cited by 2 publications

References 41 publications

Protective potential of SCF for mice preimplantation embryos cultured in vitro in suboptimal conditions

Protective potential of SCF for mice preimplantation embryos cultured in vitro in suboptimal conditions

Characteristics of the cell membrane fluidity, actin fibers, and mitochondrial dysfunctions of frozen‐thawed two‐cell mouse embryos

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