Oxygen consumption of single bovine embryos probed by scanning electrochemical microscopy

被引:158
作者
Shiku, H
Shiraishi, T
Ohya, H
Matsue, T
Abe, H
Hoshi, H
Kobayashi, M
机构
[1] Japan Sci & Technol Corp, Reg Joint Res Project Yamagata Prefecture, Yamagata 9902473, Japan
[2] Inst Life Support Technol, Yamagata 9902473, Japan
[3] Tohoku Univ, Grad Sch Engn, Dept Biomol Engn, Sendai, Miyagi 9808579, Japan
[4] Res Inst Funct Peptides, Yamagata 9900823, Japan
[5] Yamagata Agr Res & Training Ctr, Shinjo 9960041, Japan
关键词
D O I
10.1021/ac010339j
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Oxygen consumption of individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). A probe microelectrode was used to scan near a single embryo surface in a culture medium to monitor the oxygen reduction current at 37 degreesC, under a water-saturated atmosphere of 5% CO2 and 95% air. The oxygen concentration profiles near the embryos were in good agreement with the theoretical spherical diffusion. When an embryo reached the stage of a morula with a 74-mum radius on day 6 after in Nitro fertilization, the oxygen concentration difference (DeltaC) between the bulk solution and the morula surface was 6.90 +/- 1.35 muM. The oxygen consumption rate (F) of the single morula was estimated to be (1.40 +/- 0.27) x 10(-14) Mol s(-1). After the SECM measurement, the embryo was continuously cultured for another 2 days and grew to the stage of a blastocyst with a 100-mum radius. For the blastocyst, the DeltaC values for the inner cell mass side and the trophoblast side were 16.40 +/- 1.83 and 9.14 +/- 1.68 muM, respectively. The oxygen consumption rate of the blastocyst was found to be in the range of (2.50 +/- 0.46) x 10(-14) Mol S-1 < F < (4.49 +/- 0.50) x 10(-14) Mol s(-1). We have carried out SECM measurements for 19 embryos, and the results were compared in detail with these from an optical microscopic observation. The DeltaC values for the morulae on day 6 after in vitro fertilization were strongly related to the morphological embryo quality. The morulae showing a larger DeltaC value developed into blastocysts of a larger size, and the DeltaC value after the subsequent 2 days of cultivation was found to be increased.
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收藏
页码:3751 / 3758
页数:8
相关论文
共 53 条
  • [21] 2-I
  • [22] DETERMINATION OF PENTOSE-PHOSPHATE AND EMBDEN-MEYERHOF PATHWAY ACTIVITIES IN BOVINE EMBRYOS
    JAVED, MH
    WRIGHT, RW
    [J]. THERIOGENOLOGY, 1991, 35 (05) : 1029 - 1037
  • [23] Oxygen microsensor and its application to single cells and mouse pancreatic islets
    Jung, SK
    Gorski, W
    Aspinwall, CA
    Kauri, LM
    Kennedy, RT
    [J]. ANALYTICAL CHEMISTRY, 1999, 71 (17) : 3642 - 3649
  • [24] KAYA T, UNPUB
  • [25] Energy metabolism in preimplantation bovine embryos derived in vitro or in vivo
    Khurana, NK
    Niemann, H
    [J]. BIOLOGY OF REPRODUCTION, 2000, 62 (04) : 847 - 856
  • [26] Land SC, 1999, J EXP BIOL, V202, P211
  • [27] APPLICATION OF SCANNING ELECTROCHEMICAL MICROSCOPY TO BIOLOGICAL SAMPLES
    LEE, CM
    KWAK, JY
    BARD, AJ
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (05) : 1740 - 1743
  • [28] LEESE HJ, 1984, J REPROD FERTIL, V72, P9, DOI 10.1530/jrf.0.0720009
  • [29] LESZCZYSZYN DJ, 1990, J BIOL CHEM, V265, P14736
  • [30] BOVINE EMBRYO MORPHOLOGY AND EVALUATION
    LINDNER, GM
    WRIGHT, RW
    [J]. THERIOGENOLOGY, 1983, 20 (04) : 407 - 416