Abstract
A “precocious” cleavage furrow develops and ingresses during early prometaphase in Mesostoma ehrenbergii spermatocytes (Forer and Pickett-Heaps Eur J Cell Biol 89:607-618, 2010). In response to chromosome movements which regularly occur during prometaphase and that alter the balance of chromosomes in the two half-spindles, the precocious furrow shifts its position along the cell, moving 2–3 μm towards the half cell with fewer chromosomes (Ferraro-Gideon et al. Cell Biol Int 37:892-898, 2013). This process continues until proper segregation is achieved and the cell enters anaphase with the cleavage furrow again in the middle of the cell. At anaphase, the furrow recommences ingression. Spindle microtubules (MTs) are implicated in various furrow positioning models, and our experiments studied the responses of the precocious furrows to the absence of spindle MTs. We depolymerized spindle MTs during prometaphase using various concentrations of nocodazole (NOC) and colcemid. The expected result is that the furrow should regress and chromosomes remain in the midzone of the cell (Cassimeris et al. J Cell Sci 96:9-15, 1990). Instead, the furrows commenced ingression and all three bivalent chromosomes moved to one pole while the univalent chromosomes, that usually reside at the two poles, either remained at their poles or moved to the opposite pole along with the bivalents, as described elsewhere (Fegaras and Forer 2018). The microtubules were completely depolymerized by the drugs, as indicated by immunofluorescence staining of treated cells (Fegaras and Forer 2018), and in the absence of microtubules, the furrows often ingressed (in 33/61 cells) at a rate similar to normal anaphase ingression (~ 1 μm/min), while often simultaneously moving toward one pole. Thus, these results indicate that in the absence of anaphase and of spindle microtubules, cleavage furrows resume ingression.
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This work was supported by research grants to AF from the Natural Sciences and Engineering Research Council of Canada.
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Supplementary video 1
cell treated with 20 μM NOC between 1259:54 and 13:00:01, when the images become out of focus. The bivalents oscillated up and back to the two spindle poles initially. After NOC was added, the oscillations ceased as the bivalents stretched out. The bivalent kinetochores facing the upper pole detached at around 13:01:16 and moved into the bottom half-spindle, leaving the univalents behind at the top pole. After these movements the cleavage furrow slowly ingressed and simultaneously moved towards the upper pole, forming a cell with two unequal parts, that looks like a ‘shmoo’. (MOV 6899 kb)
Supplementary video 2
cell treated with 20 μM NOC between frames 91–95, when the image was out of focus. [The time between each frame is 2 s.] As in video 1, after NOC the chromosomes stretched out; all bivalent kinetochores detached from the upper pole, starting at about frame 115, and moved to the bottom half-spindle together with the univalent from the upper pole. Subsequent to the chromosome movements the cleavage furrow ingressed and moved towards the upper pole, ending up at the cell periphery. (MOV 5514 kb)
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Fegaras, E., Forer, A. Precocious cleavage furrows simultaneously move and ingress when kinetochore microtubules are depolymerized in Mesostoma ehrenbergii spermatocytes. Protoplasma 255, 1401–1411 (2018). https://doi.org/10.1007/s00709-018-1239-8
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DOI: https://doi.org/10.1007/s00709-018-1239-8