scholarly journals Oscillatory movements of monooriented chromosomes and their position relative to the spindle pole result from the ejection properties of the aster and half-spindle.

1986 ◽  
Vol 103 (2) ◽  
pp. 581-591 ◽  
Author(s):  
C L Rieder ◽  
E A Davison ◽  
L C Jensen ◽  
L Cassimeris ◽  
E D Salmon

During mitosis a monooriented chromosome oscillates toward and away from its associated spindle pole and may be positioned many micrometers from the pole at the time of anaphase. We tested the hypothesis of Pickett-Heaps et al. (Pickett-Heaps, J. D., D. H. Tippit, and K. R. Porter, 1982, Cell, 29:729-744) that this behavior is generated by the sister kinetochores of a chromosome interacting with, and moving in opposite direction along, the same set of polar microtubules. When the sister chromatids of a monooriented chromosome split at the onset of anaphase in newt lung cells, the proximal chromatid remains stationary or moves closer to the pole, with the kinetochore leading. During this time the distal chromatid moves a variable distance radially away from the pole, with one or both chromatid arms leading. Subsequent electron microscopy of these cells revealed that the kinetochore on the distal chromatid is free of microtubules. These results suggest that the distal kinetochore is not involved in the positioning of a monooriented chromosome relative to the spindle pole or in its oscillatory movements. To test this conclusion we used laser microsurgery to create monooriented chromosomes containing one kinetochore. Correlative light and electron microscopy revealed that chromosomes containing one kinetochore continue to undergo normal oscillations. Additional observations on normal and laser-irradiated monooriented chromosomes indicated that the chromosome does not change shape, and that the kinetochore region is not deformed, during movement away from the pole. Thus movement away from the pole during an oscillation does not appear to arise from a push generated by the single pole-facing kinetochore fiber, as postulated (Bajer, A. S., 1982, J. Cell Biol., 93:33-48). When the chromatid arms of a monooriented chromosome are cut free of the kinetochore, they are immediately ejected radially outward from the spindle pole at a constant velocity of 2 micron/min. This ejection velocity is similar to that of the outward movement of an oscillating chromosome. We conclude that the oscillations of a monooriented chromosome and its position relative to the spindle pole result from an imbalance between poleward pulling forces acting at the proximal kinetochore and an ejection force acting along the chromosome, which is generated within the aster and half-spindle.

1983 ◽  
Vol 97 (1) ◽  
pp. 6-14 ◽  
Author(s):  
P Cancalon

Degeneration was followed in the garfish olfactory nerve after removal of the mucosa containing the cell bodies. Degeneration, as measured by a decrease in the weight of consecutive 3-mm nerve segments, spreads at constant velocity from the site of injury toward the synaptic area. The proximodistal degeneration is temperature dependent and progresses from 0.3 mm/d at 10 degrees C to 13.0 mm/d at 35 degrees C. Between 14 and 35 degrees C, the velocity increases linearly with temperature. At all the temperatures investigated, these proximodistal degeneration velocities are identical to the rates of slow intraaxonal flow measured in axons detached from their cell bodies, or to the rates measured in regenerating fibers, and, except at 10 degrees C, are 3.3 times faster than the rate of slow flow in intact nerves. These results were confirmed by light and electron microscopy. We hypothesize that the collapse and subsequent degeneration of the axons is the result of a proximodistal depletion of cytoskeletal elements no longer provided by the cell body to the axon by slow intraaxonal flow. A significant number of axons disappeared rapidly from the nerve before the arrival of the slow degenerative wave. From studies by other groups, this rapid degeneration may be the result of a lack of rapidly transported, mainly membranous components.


1972 ◽  
Vol 55 (2) ◽  
pp. 368-389 ◽  
Author(s):  
James R. Aist ◽  
P. H. Williams

Mitosis in Fusarium oxysporum Schlect. was studied by light and electron microscopy. The average times required for the stages of mitosis, as determined from measurements made on living nuclei, were as follows: prophase, 70 sec; metaphase, 120 sec; anaphase, 13 sec; and telophase, 125 sec, for a total of 5.5 min. New postfixation procedures were developed specifically to preserve the fine-structure of the mitotic apparatus. Electron microscopy of mitotic nuclei revealed a fibrillo-granular, extranuclear Spindle Pole Body (SPB) at each pole of the intranuclear, microtubular spindles. Metaphase chromosomes were attached to spindle microtubules via kinetochores, which were found near the spindle poles at telophase. The still-intact, original nuclear envelope constricted around the incipient daughter nuclei during telophase.


2018 ◽  
Vol 29 (11) ◽  
pp. 1332-1345 ◽  
Author(s):  
Anna H. Klemm ◽  
Agneza Bosilj ◽  
Matko Gluncˇic´ ◽  
Nenad Pavin ◽  
Iva M. Tolic´

During metaphase, sister chromatids are connected to microtubules extending from the opposite spindle poles via kinetochores to protein complexes on the chromosome. Kinetochores congress to the equatorial plane of the spindle and oscillate around it, with kinesin-8 motors restricting these movements. Yet, the physical mechanism underlying kinetochore movements is unclear. We show that kinetochore movements in the fission yeast Schizosaccharomyces pombe are regulated by kinesin-8-promoted microtubule catastrophe, force-induced rescue, and microtubule dynamic instability. A candidate screen showed that among the selected motors only kinesin-8 motors Klp5/Klp6 are required for kinetochore centering. Kinesin-8 accumulates at the end of microtubules, where it promotes catastrophe. Laser ablation of the spindle resulted in kinetochore movement toward the intact spindle pole in wild-type and klp5Δ cells, suggesting that kinetochore movement is driven by pulling forces. Our theoretical model with Langevin description of microtubule dynamic instability shows that kinesin-8 motors are required for kinetochore centering, whereas sensitivity of rescue to force is necessary for the generation of oscillations. We found that irregular kinetochore movements occur for a broader range of parameters than regular oscillations. Thus, our work provides an explanation for how regulation of microtubule dynamic instability contributes to kinetochore congression and the accompanying movements around the spindle center.


1977 ◽  
Vol 72 (2) ◽  
pp. 368-379 ◽  
Author(s):  
S Brenner ◽  
A Branch ◽  
S Meredith ◽  
M W Berns

Light and electron microscopy were used to study somatic cell reduction division occurring spontaneously in tetraploid populations of rat kangaroo Potorous tridactylis (PtK2) cells in vitro. Light microscopy coupled with time-lapse photography documented the pattern of reduction division which includes an anaphase-like movement of double chromatid chromosomes to opposite spindle poles followed by the organization of two separate metaphase plates and synchronous anaphase division to form four poles and four daughter nuclei. The resulting daughter cells were isolated and cloned, showing their viability, and karyotyped to determine their ploidy. Ultrastructural analysis of cells undergoing reduction consistently revealed two duplexes of centrioles (one at each of two spindle poles) and two spindle poles in each cell that lacked centrioles but with microtubules terminating in a pericentriolar-like cloud of material. These results suggest that the centriole is not essential for spindle pole formation and division and implicate the could region as a necessary component of the spindle apparatus.


1982 ◽  
Vol 28 (9) ◽  
pp. 1059-1077 ◽  
Author(s):  
M.-L. Ashton ◽  
P. B. Moens

Conjugation in Schizosaccharomyces octosporus is described through the use of interference contrast microscopy, fluorescence microscopy, and electron microscopy of serial sections. At the light microscope level, mating was frequently observed to occur between cells of common ancestry. Fluorescent staining of the nuclei showed that nuclear migration occurs prior to karyogamy, and following diploidization the nucleus then migrates to the end of the cell. A brightly fluorescent spot was found at the apex of the migration nucleus. At the electron microscope level, the results showed that nuclear movement occurs in the presence of cytoplasmic microtubules that are associated with the spindle pole body, the conjugatory nuclei first fuse at or near the spindle pole bodies, and fusion of the spindle bodies occurs apparently by stacking one onto the other.


The motor innervation of cat spindles was examined in hindlimb muscles using a variety of techniques employed in light and electron microscopy. Observations were made on teased, silver preparations of 267 spindles sampled from the peroneal, flexor hallucis longus, and soleus muscles, hereafter referred to as the PER /FHL /SOL series. The y innervation . Trail endings are almost invariably present, and innervate both bag and chain muscle fibres. T rail fibres accounted for 64.6 to 74.8 % of the total fusimotor supply to samples of spindle poles in the PER/FHL /SOL series, the m ean num ber of fibres per pole varying from 2.7 to 5.0 in the different muscles, and the mean number of ramifications (areas of synaptic contact) per fibre being 3.7. By contrast, the p 2 innervation of a spindle pole generally consists of a single fibre supplying only one plate. In the above samples p 2 fibres accounted for 4.1 to 28.0% of the total fusimotor supply, and the mean number of fibres per pole varied from 0.3 to 1.2 in the different muscles. Ninety per cent of p 2 plates innervate bag fibres. The α innervation . The structure of p 1 plates as seen in both light and electron microscopy compares very closely with that of extrafusal plates. After nerve section p 1 plates degenerate at the same time as extrafusal plates, being the first of the three types of fusimotor ending to disappear. The frequency of the p x innervation is similar to that of the p 2 innervation. In the same samples of P E R /F H L /S O L spindle poles as above p x fibres accounted for 6.0 to 28.8 % of the total fusimotor supply, the mean number of fibres per pole varying from 0.25 to 2.1 in the different muscles. The majority of p 1 fibres enter a pole to terminate in one plate only. Seventy-five per cent of the plates innervate bag fibres. The three types of fusimotor ending are thus not selectively distributed to the two types of intrafusal muscle fibre. All three types of fusimotor fibre may branch within the spindle so as to innervate both bag and chain fibres. Bag fibres receive both types of plate ending as well as trail endings. Most chain fibres receive trail endings only; the rest receive either a p 1 or a p 2 plate innervation in addition, 25 % of the p 1 and 10% of the p 2 innervation being distributed to chain fibres. The significance of this nonselective innervation is interpreted as indicating that the type of contraction elicited by stimulating a fusimotor fibre depends upon the type of ending initiating it rather than upon the type of muscle fibre executing it. Reasons are given for concluding that the dynamic response is controlled via the p 1 and p 2 plates, and that the static response is controlled by the trail endings. The participation of the a fibres in mammalian fusimotor innervation, previously regarded as a vestigial feature, proved to be widespread in the muscles studied and more prevalent in fast muscles (FHL, peroneus digiti quinti) than slow (soleus). A low frequency of p 1 innervation is offset by a high frequency of p 2 (as in peroneus longus), and vice versa (as in FHL). It is unlikely that collaterals from slow a fibres innervating type B muscle fibres are wholly responsible for the high frequency of the p 1 innervation in FHL, and it is suggested that collaterals may also be derived from fast a fibres innervating type C muscle fibres. The possibility of there being some motor fibres of a conduction velocity and with an exclusively fusimotor distribution is also taken into account.


1974 ◽  
Vol 52 (11) ◽  
pp. 2323-2333 ◽  
Author(s):  
E. C. Setliff ◽  
H. C. Hoch ◽  
R. F. Patton

Nuclear division in basidia of Poria latemarginata was studied comparatively by light and electron microscopy. Premeiotic mitosis occurred in the lower half of the basidium and was oriented parallel to the longitudinal axis of the basidium. Mitosis was not observed with the light microscope and only late anaphase figures of mitosis were seen with the electron microscope. Mitosis was intranuclear with microtubules oriented between two spindle pole bodies (SPBs). The SPBs were spherical with a central core of material slightly more electron opaque than the surrounding SPB material. The nuclear envelope remained intact except at the SPBs.Divisions I and II of meiosis were chiastobasidial and occurred at the apices of basidia. The major features of meiosis observed by both light and electron microscopy were (1) karyogamy followed by the presence of one or two nucleoli in the prophase nucleus; (2) elongated chromosomes and synapsis at late zygotene – pachytene; (3) occurrence of a spindle at metaphase–anaphase composed of chromosomal and continuous microtubules associated with the SPBs; (4) absence of a metaphase plate with chromosomes arranged randomly around a zone of continuous microtubules; (5) condensation of chromosomes and asynchronous separation at anaphase; (6) kinetochores at anaphase; (7) the nuclear envelope remaining intact throughout meiosis except for discontinuities at the SPBs; (8) membrane-bound vesicles associated with chromosomes during division; and (9) separation of daughter nuclei at telophase. Stages of division II meiosis were observed less frequently and were similar to division I. The four postmeiotic nuclei then migrated back toward the central part of the basidium. Sterigmata developed at this time. Postmeiotic nuclei were surrounded by one or two envelopes of perinuclear endoplasmic reticulum before their migration into basidiospores. Electron-opaque inclusions occurred within the nuclei at this stage.


1990 ◽  
Vol 110 (2) ◽  
pp. 391-404 ◽  
Author(s):  
T S Hays ◽  
E D Salmon

To examine the dependence of poleward force at a kinetochore on the number of kinetochore microtubules (kMTs), we altered the normal balance in the number of microtubules at opposing homologous kinetochores in meiosis I grasshopper spermatocytes at metaphase with a focused laser microbeam. Observations were made with light and electron microscopy. Irradiations that partially damaged one homologous kinetochore caused the bivalent chromosome to shift to a new equilibrium position closer to the pole to which the unirradiated kinetochore was tethered; the greater the dose of irradiation, the farther the chromosome moved. The number of kMTs on the irradiated kinetochore decreased with severity of irradiation, while the number of kMTs on the unirradiated kinetochore remained constant and independent of chromosome-to-pole distance. Assuming a balance of forces on the chromosome at congression equilibrium, our results demonstrate that the net poleward force on a chromosome depends on the number of kMTs and the distance from the pole. In contrast, the velocity of chromosome movement showed little dependence on the number of kMTs. Possible mechanisms which explain the relationship between the poleward force at a kinetochore, the number of kinetochore microtubules, and the lengths of the kinetochore fibers at congression equilibrium include a "traction fiber model" in which poleward force producers are distributed along the length of the kinetochore fibers, or a "kinetochore motor-polar ejection model" in which force producers located at or near the kinetochore pull the chromosomes poleward along the kMTs and against an ejection force that is produced by the polar microtubule array and increases in strength toward the pole.


Author(s):  
Odell T. Minick ◽  
Hidejiro Yokoo ◽  
Fawzia Batti

Vacuolated cells in the liver of young rats were studied by light and electron microscopy following the administration of vitamin A (200 units per gram of body weight). Their characteristics were compared with similar cells found in untreated animals.In rats given vitamin A, cells with vacuolated cytoplasm were a prominent feature. These cells were found mostly in a perisinusoidal location, although some appeared to be in between liver cells (Fig. 1). Electron microscopy confirmed their location in Disse's space adjacent to the sinusoid and in recesses between liver cells. Some appeared to be bordering the lumen of the sinusoid, but careful observation usually revealed a tenuous endothelial process separating the vacuolated cell from the vascular space. In appropriate sections, fenestrations in the thin endothelial processes were noted (Fig. 2, arrow).


Author(s):  
John H. L. Watson ◽  
John L. Swedo ◽  
M. Vrandecic

The ambient temperature and the nature of the storage fluids may well have significant effects upon the post-implantation behavior of venus autografts. A first step in the investigation of such effects is reported here. Experimental conditions have been set which approximate actual operating room procedures. Saphenous veins from dogs have been used as models in the experiments. After removal from the dogs the veins were kept for two hours under four different experimental conditions, viz at either 4°C or 23°C in either physiological saline or whole canine arterial blood. At the end of the two hours they were prepared for light and electron microscopy. Since no obvious changes or damage could be seen in the veins by light microscopy, even with the advantage of tissue specific stains, it was essential that the control of parameters for successful grafts be set by electron microscopy.


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