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Golgi clusters and vesicles mediate mitotic inheritance independently of the endoplasmic reticulum.

Jokitalo E, Cabrera-Poch N, Warren G, Shima DT - J. Cell Biol. (2001)

Bottom Line: Presley, T.H.Roberts, E.Cell. 99:589-601) and suggest that these results, in part, are the consequence of slow or abortive folding of GFP-Golgi chimeras in the ER.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biotechnology, Electron Microscopy Unit, University of Helsinki, 00014 Helsinki, Finland.

ABSTRACT
We have examined the fate of Golgi membranes during mitotic inheritance in animal cells using four-dimensional fluorescence microscopy, serial section reconstruction of electron micrographs, and peroxidase cytochemistry to track the fate of a Golgi enzyme fused to horseradish peroxidase. All three approaches show that partitioning of Golgi membranes is mediated by Golgi clusters that persist throughout mitosis, together with shed vesicles that are often found associated with spindle microtubules. We have been unable to find evidence that Golgi membranes fuse during the later phases of mitosis with the endoplasmic reticulum (ER) as a strategy for Golgi partitioning (Zaal, K.J., C.L. Smith, R.S. Polishchuk, N. Altan, N.B. Cole, J. Ellenberg, K. Hirschberg, J.F. Presley, T.H. Roberts, E. Siggia, et al. 1999. Cell. 99:589-601) and suggest that these results, in part, are the consequence of slow or abortive folding of GFP-Golgi chimeras in the ER. Furthermore, we show that accurate partitioning is accomplished early in mitosis, by a process of cytoplasmic redistribution of Golgi fragments and vesicles yielding a balance of Golgi membranes on either side of the metaphase plate before cell division.

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Related in: MedlinePlus

3D reconstruction of Golgi clusters in NAGFP-PtK cells at different stages of mitosis. Four mitotic cells were sectioned completely and a series of approximately evenly spaced sections (total in parentheses) chosen to assemble a complete cell: prometaphase (20 sections); metaphase (11); anaphase (10); and telophase (18). Chromosomes (lilac) and Golgi clusters (yellow) were identified morphologically and traced, and the traces were assembled into a 3D stack. Shown is a 2D projection of the reconstructed images. An animated sequence (Video 6) is available at http://www.jcb.org/cgi/content/full/200104073/DC1, allowing the images to be rotated. Note that mitotic Golgi clusters persist throughout mitosis and are organized in the perispindle region of PtK1 cells during the metaphase to telophase period. Bar, 5 μm.
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fig5: 3D reconstruction of Golgi clusters in NAGFP-PtK cells at different stages of mitosis. Four mitotic cells were sectioned completely and a series of approximately evenly spaced sections (total in parentheses) chosen to assemble a complete cell: prometaphase (20 sections); metaphase (11); anaphase (10); and telophase (18). Chromosomes (lilac) and Golgi clusters (yellow) were identified morphologically and traced, and the traces were assembled into a 3D stack. Shown is a 2D projection of the reconstructed images. An animated sequence (Video 6) is available at http://www.jcb.org/cgi/content/full/200104073/DC1, allowing the images to be rotated. Note that mitotic Golgi clusters persist throughout mitosis and are organized in the perispindle region of PtK1 cells during the metaphase to telophase period. Bar, 5 μm.

Mentions: PtK-1 cells were seeded at low density to provide small well-separated colonies. Those containing a suitable mitotic cell were selected by phase contrast microscopy, fixed, and processed for epon embedding. Serial thin sections were cut parallel to the coverslip and sections at approximately equal intervals were photographed. Chromosomes and clusters were identified and traced on overlays that were then assembled using NIH Image. Supplemental data illustrate some of the structures chosen to assemble the final images (Supplemental Fig. S1) and an animation (Video 6) is included that permits rotation of the assembled images for easier appreciation of the individual structures. A 2D image is presented in Fig. 5 .


Golgi clusters and vesicles mediate mitotic inheritance independently of the endoplasmic reticulum.

Jokitalo E, Cabrera-Poch N, Warren G, Shima DT - J. Cell Biol. (2001)

3D reconstruction of Golgi clusters in NAGFP-PtK cells at different stages of mitosis. Four mitotic cells were sectioned completely and a series of approximately evenly spaced sections (total in parentheses) chosen to assemble a complete cell: prometaphase (20 sections); metaphase (11); anaphase (10); and telophase (18). Chromosomes (lilac) and Golgi clusters (yellow) were identified morphologically and traced, and the traces were assembled into a 3D stack. Shown is a 2D projection of the reconstructed images. An animated sequence (Video 6) is available at http://www.jcb.org/cgi/content/full/200104073/DC1, allowing the images to be rotated. Note that mitotic Golgi clusters persist throughout mitosis and are organized in the perispindle region of PtK1 cells during the metaphase to telophase period. Bar, 5 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2150754&req=5

fig5: 3D reconstruction of Golgi clusters in NAGFP-PtK cells at different stages of mitosis. Four mitotic cells were sectioned completely and a series of approximately evenly spaced sections (total in parentheses) chosen to assemble a complete cell: prometaphase (20 sections); metaphase (11); anaphase (10); and telophase (18). Chromosomes (lilac) and Golgi clusters (yellow) were identified morphologically and traced, and the traces were assembled into a 3D stack. Shown is a 2D projection of the reconstructed images. An animated sequence (Video 6) is available at http://www.jcb.org/cgi/content/full/200104073/DC1, allowing the images to be rotated. Note that mitotic Golgi clusters persist throughout mitosis and are organized in the perispindle region of PtK1 cells during the metaphase to telophase period. Bar, 5 μm.
Mentions: PtK-1 cells were seeded at low density to provide small well-separated colonies. Those containing a suitable mitotic cell were selected by phase contrast microscopy, fixed, and processed for epon embedding. Serial thin sections were cut parallel to the coverslip and sections at approximately equal intervals were photographed. Chromosomes and clusters were identified and traced on overlays that were then assembled using NIH Image. Supplemental data illustrate some of the structures chosen to assemble the final images (Supplemental Fig. S1) and an animation (Video 6) is included that permits rotation of the assembled images for easier appreciation of the individual structures. A 2D image is presented in Fig. 5 .

Bottom Line: Presley, T.H.Roberts, E.Cell. 99:589-601) and suggest that these results, in part, are the consequence of slow or abortive folding of GFP-Golgi chimeras in the ER.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biotechnology, Electron Microscopy Unit, University of Helsinki, 00014 Helsinki, Finland.

ABSTRACT
We have examined the fate of Golgi membranes during mitotic inheritance in animal cells using four-dimensional fluorescence microscopy, serial section reconstruction of electron micrographs, and peroxidase cytochemistry to track the fate of a Golgi enzyme fused to horseradish peroxidase. All three approaches show that partitioning of Golgi membranes is mediated by Golgi clusters that persist throughout mitosis, together with shed vesicles that are often found associated with spindle microtubules. We have been unable to find evidence that Golgi membranes fuse during the later phases of mitosis with the endoplasmic reticulum (ER) as a strategy for Golgi partitioning (Zaal, K.J., C.L. Smith, R.S. Polishchuk, N. Altan, N.B. Cole, J. Ellenberg, K. Hirschberg, J.F. Presley, T.H. Roberts, E. Siggia, et al. 1999. Cell. 99:589-601) and suggest that these results, in part, are the consequence of slow or abortive folding of GFP-Golgi chimeras in the ER. Furthermore, we show that accurate partitioning is accomplished early in mitosis, by a process of cytoplasmic redistribution of Golgi fragments and vesicles yielding a balance of Golgi membranes on either side of the metaphase plate before cell division.

Show MeSH
Related in: MedlinePlus