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Title: Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos

Abstract

Centrosomes, the main microtubule-organizing centers (MTOCs) in most animal cells, are important for many cellular activities such as assembly of the mitotic spindle, establishment of cell polarity, and cell movement. In nuclear transfer (NT), MTOCs that are located at the poles of the meiotic spindle are removed from the recipient oocyte, while the centrosome of the donor cell is introduced. We used mouse MII oocytes as recipients, mouse fibroblasts, rat fibroblasts, or pig granulosa cells as donor cells to construct intraspecies and interspecies nuclear transfer embryos in order to observe centrosome dynamics and functions. Three antibodies against centrin, {gamma}-tubulin, and NuMA, respectively, were used to stain the centrosome. Centrin was not detected either at the poles of transient spindles or at the poles of first mitotic spindles. {gamma}-tubulin translocated into the two poles of the transient spindles, while no accumulated {gamma}-tubulin aggregates were detected in the area adjacent to the two pseudo-pronuclei. At first mitotic metaphase, {gamma}-tubulin was translocated to the spindle poles. The distribution of {gamma}-tubulin was similar in mouse intraspecies and rat-mouse interspecies embryos. The NuMA antibody that we used can recognize porcine but not murine NuMA protein, so it was used to trace the NuMA protein ofmore » donor cell in reconstructed embryos. In the pig-mouse interspecies reconstructed embryos, NuMA concentrated between the disarrayed chromosomes soon after activation and translocated to the transient spindle poles. NuMA then immigrated into pseudo-pronuclei. After pseudo-pronuclear envelope breakdown, NuMA was located between the chromosomes and then translocated to the spindle poles of first mitotic metaphase. {gamma}-tubulin antibody microinjection resulted in spindle disorganization and retardation of the first cell division. NuMA antibody microinjection also resulted in spindle disorganization. Our findings indicate that (1) the donor cell centrosome, defined as pericentriolar material surrounding a pair of centrioles, is degraded in the 1-cell reconstituted embryos after activation; (2) components of donor cell centrosomes contribute to the formation of the transient spindle and normal functional mitotic spindle, although the contribution of centrosomal material stored in the recipient ooplasm is not excluded; and (3) components of donor cell centrosomes involved in spindle assembly may not be species-specific.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [2];  [1]
  1. State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080 (China)
  2. Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri-Columbia, MO 65211 (United States)
Publication Date:
OSTI Identifier:
20717596
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research
Additional Journal Information:
Journal Volume: 306; Journal Issue: 1; Other Information: DOI: 10.1016/j.yexcr.2005.02.009; PII: S0014-4827(05)00048-0; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIBODIES; CELL NUCLEI; CHROMOSOMES; EMBRYOS; FIBROBLASTS; MICE; MICROTUBULES; MITOSIS; OOCYTES; PROTEINS; RATS; SWINE

Citation Formats

Zhisheng, Zhong, Gang, Zhang, Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014, Xiaoqian, Meng, Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014, Yanling, Zhang, Dayuan, Chen, Schatten, Heide, and Qingyuan, Sun. Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos. United States: N. p., 2005. Web. doi:10.1016/j.yexcr.2005.02.009.
Zhisheng, Zhong, Gang, Zhang, Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014, Xiaoqian, Meng, Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014, Yanling, Zhang, Dayuan, Chen, Schatten, Heide, & Qingyuan, Sun. Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos. United States. https://doi.org/10.1016/j.yexcr.2005.02.009
Zhisheng, Zhong, Gang, Zhang, Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014, Xiaoqian, Meng, Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014, Yanling, Zhang, Dayuan, Chen, Schatten, Heide, and Qingyuan, Sun. 2005. "Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos". United States. https://doi.org/10.1016/j.yexcr.2005.02.009.
@article{osti_20717596,
title = {Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos},
author = {Zhisheng, Zhong and Gang, Zhang and Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014 and Xiaoqian, Meng and Key Laboratory of Animal Resistance, College of Life Science, Shandong Normal University, Jinan, 250014 and Yanling, Zhang and Dayuan, Chen and Schatten, Heide and Qingyuan, Sun},
abstractNote = {Centrosomes, the main microtubule-organizing centers (MTOCs) in most animal cells, are important for many cellular activities such as assembly of the mitotic spindle, establishment of cell polarity, and cell movement. In nuclear transfer (NT), MTOCs that are located at the poles of the meiotic spindle are removed from the recipient oocyte, while the centrosome of the donor cell is introduced. We used mouse MII oocytes as recipients, mouse fibroblasts, rat fibroblasts, or pig granulosa cells as donor cells to construct intraspecies and interspecies nuclear transfer embryos in order to observe centrosome dynamics and functions. Three antibodies against centrin, {gamma}-tubulin, and NuMA, respectively, were used to stain the centrosome. Centrin was not detected either at the poles of transient spindles or at the poles of first mitotic spindles. {gamma}-tubulin translocated into the two poles of the transient spindles, while no accumulated {gamma}-tubulin aggregates were detected in the area adjacent to the two pseudo-pronuclei. At first mitotic metaphase, {gamma}-tubulin was translocated to the spindle poles. The distribution of {gamma}-tubulin was similar in mouse intraspecies and rat-mouse interspecies embryos. The NuMA antibody that we used can recognize porcine but not murine NuMA protein, so it was used to trace the NuMA protein of donor cell in reconstructed embryos. In the pig-mouse interspecies reconstructed embryos, NuMA concentrated between the disarrayed chromosomes soon after activation and translocated to the transient spindle poles. NuMA then immigrated into pseudo-pronuclei. After pseudo-pronuclear envelope breakdown, NuMA was located between the chromosomes and then translocated to the spindle poles of first mitotic metaphase. {gamma}-tubulin antibody microinjection resulted in spindle disorganization and retardation of the first cell division. NuMA antibody microinjection also resulted in spindle disorganization. Our findings indicate that (1) the donor cell centrosome, defined as pericentriolar material surrounding a pair of centrioles, is degraded in the 1-cell reconstituted embryos after activation; (2) components of donor cell centrosomes contribute to the formation of the transient spindle and normal functional mitotic spindle, although the contribution of centrosomal material stored in the recipient ooplasm is not excluded; and (3) components of donor cell centrosomes involved in spindle assembly may not be species-specific.},
doi = {10.1016/j.yexcr.2005.02.009},
url = {https://www.osti.gov/biblio/20717596}, journal = {Experimental Cell Research},
issn = {0014-4827},
number = 1,
volume = 306,
place = {United States},
year = {Sun May 15 00:00:00 EDT 2005},
month = {Sun May 15 00:00:00 EDT 2005}
}