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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

Labeling of ER fragments using peroxidase cytochemistry of HRP-KDEL in NRK cells. (A) An interphase cell showing that HRP-KDEL is restricted to the ER and nuclear envelope (open arrows). The Golgi apparatus is not stained (white arrows). Metaphase cells at high (B) and low (C) magnification show extensive staining of ER fragments but no staining of Golgi clusters (large arrow in B) or vesicles (small arrows in B), some of which appear to be associated with spindle microtubules (note the centrioles upper left). The low magnification view of an anaphase cell (D) also shows extensive staining of ER fragments but no staining of Golgi clusters (arrow, far right and inset). Bars, 1 μm.
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fig8: Labeling of ER fragments using peroxidase cytochemistry of HRP-KDEL in NRK cells. (A) An interphase cell showing that HRP-KDEL is restricted to the ER and nuclear envelope (open arrows). The Golgi apparatus is not stained (white arrows). Metaphase cells at high (B) and low (C) magnification show extensive staining of ER fragments but no staining of Golgi clusters (large arrow in B) or vesicles (small arrows in B), some of which appear to be associated with spindle microtubules (note the centrioles upper left). The low magnification view of an anaphase cell (D) also shows extensive staining of ER fragments but no staining of Golgi clusters (arrow, far right and inset). Bars, 1 μm.

Mentions: These conclusions were corroborated using NRK cells that had been transiently transfected with an HRP construct tagged with the ER salvage signal, KDEL (Fig. 8) (Munro and Pelham, 1987; Connolly et al., 1994). In interphase cells the staining was restricted to the ER, intermediates on the ER to Golgi pathway (ERGIC) and, to a limited extent, cis-Golgi elements, especially when the expression level was high. During mitosis, the ER underwent fragmentation and vesiculation, but at all stages of mitosis, the pattern was very different from that observed using the SialylT-HRP construct (compare Figs. 7 and 8). Golgi clusters were readily visible and unstained, as were those vesicles found in association with the mitotic spindle microtubules, emphasizing the likelihood that these vesicles were Golgi- not ER-derived and likely involved in the equilibration process leading to accurate Golgi partitioning.


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)

Labeling of ER fragments using peroxidase cytochemistry of HRP-KDEL in NRK cells. (A) An interphase cell showing that HRP-KDEL is restricted to the ER and nuclear envelope (open arrows). The Golgi apparatus is not stained (white arrows). Metaphase cells at high (B) and low (C) magnification show extensive staining of ER fragments but no staining of Golgi clusters (large arrow in B) or vesicles (small arrows in B), some of which appear to be associated with spindle microtubules (note the centrioles upper left). The low magnification view of an anaphase cell (D) also shows extensive staining of ER fragments but no staining of Golgi clusters (arrow, far right and inset). Bars, 1 μm.
© Copyright Policy
Related In: Results  -  Collection

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

fig8: Labeling of ER fragments using peroxidase cytochemistry of HRP-KDEL in NRK cells. (A) An interphase cell showing that HRP-KDEL is restricted to the ER and nuclear envelope (open arrows). The Golgi apparatus is not stained (white arrows). Metaphase cells at high (B) and low (C) magnification show extensive staining of ER fragments but no staining of Golgi clusters (large arrow in B) or vesicles (small arrows in B), some of which appear to be associated with spindle microtubules (note the centrioles upper left). The low magnification view of an anaphase cell (D) also shows extensive staining of ER fragments but no staining of Golgi clusters (arrow, far right and inset). Bars, 1 μm.
Mentions: These conclusions were corroborated using NRK cells that had been transiently transfected with an HRP construct tagged with the ER salvage signal, KDEL (Fig. 8) (Munro and Pelham, 1987; Connolly et al., 1994). In interphase cells the staining was restricted to the ER, intermediates on the ER to Golgi pathway (ERGIC) and, to a limited extent, cis-Golgi elements, especially when the expression level was high. During mitosis, the ER underwent fragmentation and vesiculation, but at all stages of mitosis, the pattern was very different from that observed using the SialylT-HRP construct (compare Figs. 7 and 8). Golgi clusters were readily visible and unstained, as were those vesicles found in association with the mitotic spindle microtubules, emphasizing the likelihood that these vesicles were Golgi- not ER-derived and likely involved in the equilibration process leading to accurate Golgi partitioning.

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