Alteration of Spindle Formation and Chromosome Alignment in Post-Ovulatory Aging of Mouse Oocytes
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초록
The objective of this study was to elucidate the dynamics of microtubules in post-ovulatory aging in vivo and in vitro of mouse oocytes. The fresh ovulated oocytes were obtained from oviducts of superovulated female ICR mice at 16 hours after hCG injection. The post-ovulatory aged oocytes were collected at 24 and 48 hours after hCG injection from in vivo and in vitro, respectively. Immunocytochemistry was performed on β-tubulin and acetylated α-tubulin. The microtubules were localized in the spindle assembly, which was barrel-shaped or slightly pointed at its poles and located peripherally in the fresh ovulated oocytes. The frequency of misaligned metaphase chromosomes were significantly increased in post-ovulatory aged oocytes after 48 hours of hCG injection. The spindle length and width of post-ovulatory aged oocytes were significantly different from those of fresh ovulated oocytes, respectively. The staining intensity of acetylated α-tubulin showed stronger in post-ovulatory aged oocytes than that in the fresh ovulated oocytes. In the aged oocytes, the spindles had moved towards the center of the oocytes from their original peripheral position and elongated, compared with the fresh ovulated oocytes. Microtubule organizing centers were formed and observed in the cytoplasm of the aged oocytes. On the contrary, it was not observed in the fresh ovulated oocytes. The alteration of spindle formation and chromosomes alignment substantiates the poor development and the increase of disorders from the post-ovulatory aged oocytes. It might be important to fertilize on time in ovulated oocytes for the developmental competence of embryos with normal karyotypes
Abstract
The objective of this study was to elucidate the dynamics of microtubules in post-ovulatory aging in vivo and in vitro of mouse oocytes. The fresh ovulated oocytes were obtained from oviducts of superovulated female ICR mice at 16 hours after hCG injection. The post-ovulatory aged oocytes were collected at 24 and 48 hours after hCG injection from in vivo and in vitro, respectively. Immunocytochemistry was performed on β-tubulin and acetylated α-tubulin. The microtubules were localized in the spindle assembly, which was barrel-shaped or slightly pointed at its poles and located peripherally in the fresh ovulated oocytes. The frequency of misaligned metaphase chromosomes were significantly increased in post-ovulatory aged oocytes after 48 hours of hCG injection. The spindle length and width of post-ovulatory aged oocytes were significantly different from those of fresh ovulated oocytes, respectively. The staining intensity of acetylated α-tubulin showed stronger in post-ovulatory aged oocytes than that in the fresh ovulated oocytes. In the aged oocytes, the spindles had moved towards the center of the oocytes from their original peripheral position and elongated, compared with the fresh ovulated oocytes. Microtubule organizing centers were formed and observed in the cytoplasm of the aged oocytes. On the contrary, it was not observed in the fresh ovulated oocytes. The alteration of spindle formation and chromosomes alignment substantiates the poor development and the increase of disorders from the post-ovulatory aged oocytes. It might be important to fertilize on time in ovulated oocytes for the developmental competence of embryos with normal karyotypes
참고문헌 (31)
- Bulinski JC, "Posttranslational modifications of a-tubulin: Detyrosination and acetylation differentiate populations of interphase microtubules in cultured cells", J Cell Biol, 106, pp.1213~1220 (1988)
- Diggins M, "Distribution of acetylated a-tubulin in Physarum polycephalum", J Cell Biol, 104, pp.303~309 (1987)
- Eichenlaub-Ritter U, "Alteration to the microtubular cytoskeleton and increased disorder of chromosome alignment in spontaneously ovulated mouse oocytes aged in vivo: an immunofluorecence study", Chromosoma, 94, pp.337~345 (1986)
- Huang JC, "Changes in histone acetylation during postovulatory aging of mouse oocyte", Biol Reprod, 77, pp.666~670 (2007)
- Jin YX, "Alterations of spindle and microfilament assembly in aged cat oocytes", Reprod Domest Anim, 45, pp.865~871 (2010)
- Kan R, "Regulation of mouse oocyte microtubule and organelle dynamics by PADI6 and the cytoplasmic lattices", Dev Biol, 350, pp.311~322 (2011)
- Kim NH, "Cytoskeletal alteration in aged porcine oocytes and parthenogenesis", Mol Reprod Dev, 43, pp.513~518 (1996)
- 김유나, "Immature Oocyte-specific Zap70 and Its Functional Analysis in Regulating Oocyte Maturation", 발생과 생식, 13(3), pp.145~153 (2009), 한국발생생물학회
- L'Hernault SW, "Chlamydomonas a-tubulin is posttranslationally modified in flagellar during flagella assembly", J Cell Biol, 97, pp.258~263 (1983)
- L'Hernault SW, "Reversal of the posttranslational modification on Chlamydomonas flagellar a-tubulin occurs during flagellar resortion", J Cell Biol, 100, pp.457~462 (1985)
- LeDizet M, "Cytoplasmic microtubules containing acetylated a-tubulin in Chlamydomonas reinhardtii: Spatial arrangement and properties", J Cell Biol, 103, pp.13~22 (1986)
- Ma W, "Reduced expression of MAD2, BCL2, and MAP kinase activity in pig oocytes after in vitro aging are associated with defects in sister chromatid segregation during meiosis II and embryo fragmentation after activation", Biol Reprod, 72, pp.373~383 (2005)
- Maro B, "Changes in the actin distribution during fertilization of the mouse egg", J Embryol Exp Morphol, 81, pp.211~237 (1984)
- Maro B, "Mechanism of polar body formation in the mouse oocyte: an interaction between the centrosomes, the cytoskeleton and plasma membrane", J Embryol Exp Morphol, 92, pp.11~32 (1986)
- Miao YL, "Oocyte aging: cellular and molecular changes, developmental potential and reversal possibility", Hum Reprod Update, 15, pp.573~585 (2009)
- Miao YL, "Centrosome abnormalities during porcine oocyte aging", Environ Mol Mutagen, 50, pp.666~671 (2009)
- Piperno G, "Monoclonal antibodies specific for an acetylated form of a-tubulin recognize the antigen in cilia and flagella from a variety of organisms", J Cell Biol, 101, pp.2085~2094 (1985)
- Piperno G, "Microtubules containing acetylated a-tubulin in mammalian in culture", J Cell Biol, 104, pp.289~302 (1987)
- Sasse R, "Acetylated a-tubulin in Physarum: Immunological characterization of the isotype and its usage in particular microtubular organel", J Cell Biol, 104, pp.41~49 (1987)
- Sato K, "Second meiotic division and polar body formation in mouse eggs fertilized in vitro", Gamete Res, 2, pp.282~293 (1979)
- Schatten G, "Acetylated α-tubulin in microtubules during mouse fertilization and early development", Devel Biol, 130, pp.74~86 (1988)
- Schatten G, "Microtubule configurations during fertilization, mitosis and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization", Proc Natl Acad Sci USA, 82, pp.4152~4156 (1985)
- Schulze E, "Posttranslational modification and microtubule stability", J Cell Biol, 105, pp.2167~2177 (1987)
- Simerly C, "Techniques for localization of specific molecules in oocytes and embryos", Method Enzymol, 225, pp.516~552 (1993)
- Steuerwald NM, "Postovulatory aging of mouse oocytes leads to decreased MAD2 transcripts and increased frequencies of premature centromere separation and anaphase", Mol Hum Reprod, 11, pp.623~630 (2005)
- Szollosi D, "Morphological changes in mouse eggs due to aging in the fallopian tube", Am J Anat, 130, pp.209~226 (1971)
- Szollosi D, "Absence of centrioles in the first and second meiotic spindles of mouse oocytes", J Cell Sci, 11, pp.521~541 (1972)
- Webb M, "Parthenogenesis and cytoskeleton organization in aging mouse eggs", J Embryol Exp Morphol, 95, pp.131~145 (1986)
- Webster DR, "Microtubules are acetylated in domains which turnover slowly", J Cell Sci, 92, pp.57~65 (1989)
- Wolf N, "Temporal and spatial pattern of differences in microtubule behavior during Drosophila embryogenesis revealed by distribution of a-tubulin isoform", Development, 102, pp.311~324 (1988)
- Xiong B, "BRCA1 is required for meiotic spindle assembly and spindle assembly checkpoint activation in mouse oocytes", Biol Reprod, 79, pp.718~726 (2008)