Swinging a sword: how microtubules search for their targets

The cell interior is in constant movement, which is to a large extent determined by microtubules, thin and long filaments that permeate the cytoplasm. To move large objects, microtubules need to connect them to the site of their destination. For example, during cell division, microtubules connect chromosomes with the spindle poles via kinetochores, protein complexes on the chromosomes. A general question is how microtubules, while being bound to one structure, find the target that needs to be connected to this structure. Here we review the mechanisms of how microtubules search for kinetochores, with emphasis on the recently discovered microtubule feature to explore space by pivoting around the spindle pole. In addition to accel- erating the search for kinetochores, pivoting helps the microtubules to search for cortical anchors, as well as to self-organize into parallel arrays and asters to target spe- cific regions of the cell. Thus, microtubule pivoting con- stitutes a mechanism by which they locate targets in different cellular contexts

Long-term (10 years) prognostic value of a normal thallium-201 myocardial exercise scintigraphy in patients with coronary artery disease documented by angiography

In order to assess the prognostic significance of normal exercise thallium-210 myocardial scintigraphy in patients with documented coronary artery disease, we studied the incidence of cardiac death and non-fatal myocardial infarction in 69 symptomatic patients without prior Q wave myocardial infarction, who demonstrated one or more significant coronary lesions (stenosis ≤70%) on an angiogram performed within 3 months of scintigraphy (Group 1). These patients were compared to a second group of 136 patients with an abnormal exercise scintigram, defined by the presence of reversible defect(s) and angiographically proven coronary artery disease (Group 2), and to a third group of 102 patients with normal exercise scintigraphy without significant coronary lesions (stenosis ≥30%) or with normal coronary angiography (Group 3). In contrast to coronary lesions observed in Group 2, patients in Group I presented more frequently with single- vessel disease (83% vs 35%, P>0·0001) and with more distal lesions (55% vs 23%, P>0·0001). Over a mean follow-up period of 8·6 years, one fatal and eight non-fatal cases of myocardial infarction were observed in Group 1. The majority of patients in Group 1 were treated medically: only 24 (35%) underwent myocardial revascularization, usually by coronary angioplasty. There was no significant difference in the incidence of combined major cardiac events (cardiac death, non-fatal myocardial infarction) in patients with normal exercise scintigraphy, with or without documented coronary artery disease (Groups 1 and 3), while the incidence was higher in Group 2. However, while the mortality remained very low in Group 1, the incidence of non-fatal myocardial infraction was not different from that of Group 2, where most patients underwent revascularization procedures. In conclusion, patients with coronary artery disease and a normal exercise thallium-201 myocardial scintigram usually have mild coronary lesions (single-vessel disease, distal location) and good long-term prognosis, with a low incidence of cardiac deat

A stochastic model of Min oscillations in Escherichia coli and Min protein segregation during cell division

The Min system in Escherichia coli directs division to the centre of the cell through pole-to-pole oscillations of the MinCDE proteins. We present a one dimensional stochastic model of these oscillations which incorporates membrane polymerisation of MinD into linear chains. This model reproduces much of the observed phenomenology of the Min system, including pole-to-pole oscillations of the Min proteins. We then apply this model to investigate the Min system during cell division. Oscillations continue initially unaffected by the closing septum, before cutting off rapidly. The fractions of Min proteins in the daughter cells vary widely, from 50%-50% up to 85%-15% of the total from the parent cell, suggesting that there may be another mechanism for regulating these levels in vivo.Comment: 19 pages, 12 figures (25 figure files); published at http://www.iop.org/EJ/journal/physbi

Pivot-and-bond model explains microtubule bundle formation

During mitosis, microtubules form a spindle, which is responsible for proper segregation of the genetic material. A common structural element in a mitotic spindle is a parallel bundle, consisting of two or more microtubules growing from the same origin and held together by cross-linking proteins. An interesting question is what are the physical principles underlying the formation and stability of such microtubule bundles. Here we show, by introducing the pivot-and-bond model, that random angular movement of microtubules around the spindle pole and forces exerted by cross-linking proteins can explain the formation of microtubule bundles as observed in our experiments. The model predicts that stable parallel bundles can form in the presence of either passive crosslinkers or plus-end directed motors, but not minus-end directed motors. In the cases where bundles form, the time needed for their formation depends mainly on the concentration of cross-linking proteins and the angular diffusion of the microtubule. In conclusion, the angular motion drives the alignment of microtubules, which in turn allows the cross-linking proteins to connect the microtubules into a stable bundle

Bridging the gap between sister kinetochores

The main task of the mitotic spindle is to generate forces that position the chromosomes at the metaphase plate and subsequently pull them apart toward the opposite spindle poles. These forces in living cells are, unfortunately, not easily accessible by current experimental techniques.1 Nicklas RB. J Cell Biol 1983; 97:542-8; PMID:6885908; http://dx.doi.org/10.1083/jcb.97.2.542[CrossRef], [PubMed], [Web of Science ®] However, much about the forces can be inferred from the shape of the spindle because the shape is an outcome of forces. K-fibers, which are bundles of microtubules ending at the kinetochore, are typically curved, suggesting that they are under compression. This inference contradicts the fact that sister kinetochores and thus also sister k-fibers are under tension, leaving us with a paradox about the origin of the curved shape of the spindle

Pivoting of microtubules driven by minus-end-directed motors leads to spindle assembly

Background At the beginning of mitosis, the cell forms a spindle made of microtubules and associated proteins to segregate chromosomes. An important part of spindle architecture is a set of antiparallel microtubule bundles connecting the spindle poles. A key question is how microtubules extending at arbitrary angles form an antiparallel interpolar bundle. Results Here, we show in fission yeast that microtubules meet at an oblique angle and subsequently rotate into antiparallel alignment. Our live- cell imaging approach provides a direct observation of interpolar bundle formation. By combining experiments with theory, we show that microtubules from each pole search for those from the opposite pole by performing random angular movement. Upon contact, two microtubules slide sideways along each other in a directed manner towards the antiparallel configuration. We introduce the contour length of microtubules as a measure of activity of motors that drive microtubule sliding, which we used together with observation of Cut7/kinesin-5 motors and our theory to reveal the minus-end- directed motility of this motor in vivo. Conclusion Random rotational motion helps microtubules from the opposite poles to find each other and subsequent accumulation of motors allows them to generate forces that drive interpolar bundle formation

Dynein, microtubule and cargo: a ménage à trois

To exert forces, motor proteins bind with one end to cytoskeletal filaments, such as microtubules and actin, and with the other end to the cell cortex, a vesicle or another motor. A general question is how motors search for sites in the cell where both motor ends can bind to their respective binding partners. In the present review, we focus on cytoplasmic dynein, which is required for a myriad of cellular functions in interphase, mitosis and meiosis, ranging from transport of organelles and functioning of the mitotic spindle to chromosome movements in meiotic prophase. We discuss how dynein targets sites where it can exert a pulling force on the microtubule to transport cargo inside the cell

Kinesin-8 Motors Improve Nuclear Centering by Promoting Microtubule Catastrophe

In fission yeast, microtubules push against the cell edge, thereby positioning the nucleus in the cell center. Kinesin-8 motors regulate microtubule catastrophe; however, their role in nuclear positioning is not known. Here we develop a physical model that describes how kinesin-8 motors affect nuclear centering by promoting a microtubule catastrophe. Our model predicts the improved centering of the nucleus in the presence of motors, which we confirmed experimentally in living cells. The model also predicts a characteristic time for the recentering of a displaced nucleus, which is supported by our experiments where we displaced the nucleus using optical tweezers

Compression Stockings Used During Two Soccer Matches Improve Perceived Muscle Soreness and High-Intensity Performance.

Gimenes, SV, Marocolo, M, Pavin, LN, Pagoto Spigolon, LM, Neto, OB, Côrrea da Silva, BV, Duffield, R, and Ribeiro da Mota, G. Compression stockings used during two soccer matches improve perceived muscle soreness and high-intensity performance. J Strength Cond Res XX(X): 000-000, 2018-Evidence on the use of compression stockings (CS) during soccer matches is limited. Thus, we evaluated the acute effects of CS on match-based physical performance indicators and perceptual responses during 2 consecutive soccer matches with 72-hour recovery. Twenty outfield players were randomly allocated to the CS group (20-30 mm Hg) or control group (non-CS) and performed 2 matches (5 players using CS or regular socks per team/match). Match loads {rating of perceived exertion × minutes; CS ∼830 vs. control 843 (arbitrary units [AU])} and heart rate (HR) responses (both CS and control ∼86% HRpeak) did not differ (p > 0.05) between CS and control groups. Although total distance covered did not differ (p > 0.05) between groups, CS increased distances (effect size [ES] = 0.9-1.32) in higher-speed zones (>19 km·h CS ∼550 m vs. control ∼373 m) alongside an increased number of accelerations (-50.0 to -3.0 m·s) than control (CS: 33.7 ± 11.2 vs. control: 23.8 ± 7.9; p = 0.003; ES = 1.04). Perceived recovery did not differ (p > 0.05) between groups for either match but was worse in the second match for both groups. Perceived muscle soreness increased in control after match 2 (from 3.1 ± 1.9 to 6.3 ± 1.6 AU; p < 0.0010) but did not in CS (from 2.8 ± 1.4 to 4.1 ± 1.9 AU; p = 0.6275; ES = 1.24 CS vs. control after match). Accordingly, CS use during 2 soccer matches with 72-hour recovery reduces perceived muscle soreness in the second match and increases higher-speed match running performance

Oblique circle method for measuring the curvature and twist of mitotic spindle microtubule bundles

The highly ordered spatial organization of microtubule bundles in the mitotic spindle is crucial for its proper functioning. The recent discovery of twisted shapes of microtubule bundles and spindle chirality suggests that the bundles extend along curved paths in three dimensions, rather than being confined to a plane. This, in turn, implies that rotational forces, i.e., torques, exist in the spindle in addition to the widely studied linear forces. However, studies of spindle architecture and forces are impeded by a lack of a robust method for the geometric quantification of microtubule bundles in the spindle. In this work, we describe a simple method for measuring and evaluating the shapes of microtubule bundles by characterizing them in terms of their curvature and twist. By using confocal microscopy, we obtain three-dimensional images of spindles, which allows us to trace the entire microtubule bundle. For each traced bundle, we first fit a plane and then fit a circle lying in that plane. With this robust method, we extract the curvature and twist, which represent the geometric information characteristic for each bundle. As the bundle shapes reflect the forces within them, this method is valuable for the understanding of forces that act on chromosomes during mitosis