Posterior Spindle Research Articles

The space-time profiles of sleep spindles and their coordination with slow oscillations on the electrode manifold.

Sleep spindles are important for sleep quality and cognitive functions, with their coordination with slow oscillations (SOs) potentially organizing cross-region reactivation of memory traces. Here, we describe the organization of spindles on the electrode manifold and their relation to SOs. We analyzed the sleep night EEG of 34 subjects and detected spindles and SOs separately at each electrode. We compared spindle properties (frequency, duration, and amplitude) in slow wave sleep (SWS) and Stage 2 sleep (S2); and in spindles that coordinate with SOs or are uncoupled. We identified different topographical spindle types using clustering analysis that grouped together spindles co-detected across electrodes within a short delay (±300 ms). We then analyzed the properties of spindles of each type, and coordination to SOs. We found that SWS spindles are shorter than S2 spindles, and spindles at frontal electrodes have higher frequencies in S2 compared to SWS. Furthermore, S2 spindles closely following an SO (about 10% of all spindles) show faster frequency, shorter duration, and larger amplitude than uncoupled ones. Clustering identified Global, Local, Posterior, Frontal-Right and Left spindle types. At centro-parietal locations, Posterior spindles show faster frequencies compared to other types. Furthermore, the infrequent SO-spindle complexes are preferentially recruiting Global SO waves coupled with fast Posterior spindles. Our results suggest a non-uniform participation of spindles to complexes, especially evident in S2. This suggests the possibility that different mechanisms could initiate an SO-spindle complex compared to SOs and spindles separately. This has implications for understanding the role of SOs-spindle complexes in memory reactivation.

The coordination of spindle‐positioning forces during the asymmetric division of the Caenorhabditis elegans zygote

In Caenorhabditis elegans zygote, astral microtubules generate forces essential to position the mitotic spindle, by pushing against and pulling from the cortex. Measuring microtubule dynamics there, we revealed the presence of two populations, corresponding to pulling and pushing events. It offers a unique opportunity to study, under physiological conditions, the variations of both spindle-positioning forces along space and time. We propose a threefold control of pulling force, by polarity, spindle position and mitotic progression. We showed that the sole anteroposterior asymmetry in dynein on-rate, encoding pulling force imbalance, is sufficient to cause posterior spindle displacement. The positional regulation, reflecting the number of microtubule contacts in the posterior-most region, reinforces this imbalance only in late anaphase. Furthermore, we exhibited the first direct proof that dynein processivity increases along mitosis. It reflects the temporal control of pulling forces, which strengthens at anaphase onset following mitotic progression and independently from chromatid separation. In contrast, the pushing force remains constant and symmetric and contributes to maintaining the spindle at the cell centre during metaphase.

A Novel Visualization Method for Sleep Spindles Based on Source Localization of High Density EEG.

Equivalent dipole source localization is a well-established approach to localizing the electrical activity in electroencephalogram (EEG). So far, source localization has been used primarily in localizing the epileptic source in human epileptic patients. Currently, source localization techniques have been applied to account for localizing epileptic source among the epileptic patients. Here, we present the first application of source localization in the field of sleep spindle in mouse brain. The spatial distribution of cortical potential was obtained by high density EEG and then the anterior and posterior sleep spindles were classified based on the K-mean clustering algorithm. To solve the forward problem, a realistic geometry brain model was produced based on boundary element method (BEM) using mouse MRI. Then, we applied four different source estimation algorithms (minimum norm, eLORETA, sLORETA, and LORETA) to estimate the spatial location of equivalent dipole source of sleep spindles. The estimated sources of anterior and posterior spindles were plotted in a cine-mode that revealed different topographic patterns of spindle propagation. The characterization of sleep spindles may be better be distinguished by our novel visualization method.

Characterization of topographically specific sleep spindles in mice.

Sleep spindles in humans have been classified as slow anterior and fast posterior spindles; recent findings indicate that their profiles differ according to pharmacology, pathology, and function. However, little is known about the generation mechanisms within the thalamocortical system for different types of spindles. In this study, we aim to investigate the electrophysiological behaviors of the topographically distinctive spindles within the thalamocortical system by applying high-density EEG and simultaneous thalamic LFP recordings in mice. 32-channel extracranial EEG and 2-channel thalamic LFP were recorded simultaneously in freely behaving mice to acquire spindles during spontaneous sleep. Hybrid F1 male mice of C57BL/6J and 129S4/svJae. Spindle events in each channel were detected by spindle detection algorithm, and then a cluster analysis was applied to classify the topographically distinctive spindles. All sleep spindles were successfully classified into 3 groups: anterior, posterior, and global spindles. Each spindle type showed distinct thalamocortical activity patterns regarding the extent of similarity, phase synchrony, and time lags between cortical and thalamic areas during spindle oscillation. We also found that sleep slow waves were likely to associate with all types of sleep spindles, but also that the ongoing cortical decruitment/ recruitment dynamics before the onset of spindles and their relationship with spindle generation were also variable, depending on the spindle types. Topographically specific sleep spindles show distinctive thalamocortical network behaviors.

Density and Frequency Caudo-Rostral Gradients of Sleep Spindles Recorded in the Human Cortex

This study aims at providing a quantitative description of intrinsic spindle frequency and density (number of spindles/min) in cortical areas using deep intracerebral recordings in humans. Thirteen patients suffering from pharmaco-resistant focal epilepsy and investigated through deep intracortical EEG in frontal, parietal, temporal, occipital, insular, and limbic cortices including the hippocampus were included. Spindle waves were detected from the ongoing EEG during slow wave sleep (SWS) by performing a time-frequency analysis on filtered signals (band-pass filter: 10-16 Hz). Then, spindle intrinsic frequency was determined using a fast Fourier transform, and spindle density (number of spindles per minute) was computed. Firstly, we showed that sleep spindles were recorded in all explored cortical areas, except temporal neocortex. In particular, we observed the presence of spindles during SWS in areas such as the insular cortex, medial parietal cortex, occipital cortex, and cingulate gyrus. Secondly, we demonstrated that both spindle frequency and density smoothly change along the caudo-rostral axis, from fast frequent posterior spindles to slower and less frequent anterior spindles. Thirdly, we identified the presence of spindle frequency oscillations in the hippocampus and the entorhinal cortex. Spindling activity is widespread among cortical areas, which argues for the fundamental role of spindles in cortical functions. Mechanisms of caudo-rostral gradient modulation in spindle frequency and density may result from a complex interplay of intrinsic properties and extrinsic modulation of thalamocortical and corticothalamic neurons.

Visualization of dynein-dependent microtubule gliding at the cell cortex: implications for spindle positioning

Dynein motors move along the microtubule (MT) lattice in a processive "walking" manner. In the one-cell Caenorhabditis elegans embryo, dynein is required for spindle-pulling forces during mitosis. Posteriorly directed spindle-pulling forces are higher than anteriorly directed forces, and this imbalance results in posterior spindle displacement during anaphase and an asymmetric division. To address how dynein could be asymmetrically activated to achieve posterior spindle displacement, we developed an assay to measure dynein's activity on individual MTs at the embryo cortex. Our study reveals that cortical dynein motors maintain a basal level of activity that propels MTs along the cortex, even under experimental conditions that drastically reduce anaphase spindle forces. This suggests that dynein-based MT gliding is not sufficient for anaphase spindle-pulling force. Instead, we find that this form of dynein activity is most prominent during spindle centering in early prophase. We propose a model whereby different dynein-MT interactions are used for specific spindle-positioning tasks in the one-cell embryo.

LET-99 inhibits lateral posterior pulling forces during asymmetric spindle elongation in C. elegans embryos

Cortical pulling on astral microtubules positions the mitotic spindle in response to PAR polarity cues and G protein signaling in many systems. In Caenorhabditis elegans single-cell embryos, posterior spindle displacement depends on Galpha and its regulators GPR-1/2 and LIN-5. GPR-1/2 and LIN-5 are necessary for cortical pulling forces and become enriched at the posterior cortex, which suggests that higher forces act on the posterior spindle pole compared with the anterior pole. However, the precise distribution of cortical forces and how they are regulated remains to be determined. Using spindle severing, single centrosome assays, and centrosome fragmentation, we show that both the anterior and posterior cortices generate more pulling force than the lateral-posterior region. Lateral inhibition depends on LET-99, which inhibits GPR-1/2 localization to produce a bipolar GPR-1/2 pattern. Thus, rather than two domains of cortical force, there are three. We propose that the attenuation of lateral forces prevents counterproductive pulling, resulting in a higher net force toward the posterior that contributes to spindle elongation and displacement.

A Casein Kinase 1 and PAR Proteins Regulate Asymmetry of a PIP2 Synthesis Enzyme for Asymmetric Spindle Positioning

SummarySpindle positioning is an essential feature of asymmetric cell division. The conserved PAR proteins together with heterotrimeric G proteins control spindle positioning in animal cells, but how these are linked is not known. In C. elegans, PAR protein activity leads to asymmetric spindle placement through cortical asymmetry of Gα regulators GPR-1/2. Here, we establish that the casein kinase 1 gamma CSNK-1 and a PIP2 synthesis enzyme (PPK-1) transduce PAR polarity to asymmetric Gα regulation. PPK-1 is posteriorly enriched in the one-celled embryo through PAR and CSNK-1 activities. Loss of CSNK-1 causes uniformly high PPK-1 levels, high symmetric cortical levels of GPR-1/2 and LIN-5, and increased spindle pulling forces. In contrast, knockdown of ppk-1 leads to low GPR-1/2 levels and decreased spindle forces. Furthermore, loss of CSNK-1 leads to increased levels of PIP2. We propose that asymmetric generation of PIP2 by PPK-1 directs the posterior enrichment of GPR-1/2 and LIN-5, leading to posterior spindle displacement.

Dynamic localization of LIN-5 and GPR-1/2 to cortical force generation domains during spindle positioning

G protein signaling pathways regulate mitotic spindle positioning during cell division in many systems. In Caenorhabditis elegans embryos, Gα subunits act with the positive regulators GPR-1/2 and LIN-5 to generate cortical pulling forces for posterior spindle displacement during the first asymmetric division. GPR-1/2 are asymmetrically localized at the posterior cortex by PAR polarity cues at this time. Here we show that LIN-5 colocalizes with GPR-1/2 in one-cell embryos during spindle displacement. Significantly, we also find that LIN-5 and GPR-1/2 are localized to the opposite, anterior cortex in a polarity-dependent manner during the nuclear centration and rotation movements that orient the forming spindle onto the polarity axis. The depletion of LIN-5 or GPR-1/2 results in decreased centration and rotation rates, indicating a role in force generation at this stage. The localization of LIN-5 and GPR-1/2 is largely interdependent and requires Gα. Further, LIN-5 immunoprecipitates with Gα in vivo, and this association is GPR-1/2 dependent. These results suggest that a complex of Gα/GPR-1/2/LIN-5 is asymmetrically localized in response to polarity cues, and this may be the active signaling complex that transmits asymmetries to the force generation machinery during both nuclear rotation and spindle displacement.

PAR-3 and PAR-1 Inhibit LET-99 Localization to Generate a Cortical Band Important for Spindle Positioning inCaenorhabditis elegansEmbryos

The conserved PAR proteins are localized in asymmetric cortical domains and are required for the polarized localization of cell fate determinants in many organisms. In Caenorhabditis elegans embryos, LET-99 and G protein signaling act downstream of the PARs to regulate spindle positioning and ensure asymmetric division. PAR-3 and PAR-2 localize LET-99 to a posterior cortical band through an unknown mechanism. Here we report that LET-99 asymmetry depends on cortically localized PAR-1 and PAR-4 but not on cytoplasmic polarity effectors. In par-1 and par-4 embryos, LET-99 accumulates at the entire posterior cortex, but remains at low levels at the anterior cortex occupied by PAR-3. Further, PAR-3 and PAR-1 have graded cortical distributions with the highest levels at the anterior and posterior poles, respectively, and the lowest levels of these proteins correlate with high LET-99 accumulation. These results suggest that PAR-3 and PAR-1 inhibit the localization of LET-99 to generate a band pattern. In addition, PAR-1 kinase activity is required for the inhibition of LET-99 localization, and PAR-1 associates with LET-99. Finally, examination of par-1 embryos suggests that the banded pattern of LET-99 is critical for normal posterior spindle displacement and to prevent spindle misorientation caused by cell shape constraints.

Tugging and pulling in asymmetric cell divisions

![Graphic][1] Posterior spindle displacement is delayed until metaphase by an anterior tether.In asymmetric divisions, the cell alters the normal process that centers the spindle in the mother cell, but how it regulates the timing of spindle movement is unclear. Now, on [page 245][2

Unfair Partitioning

During early development in the nematode worm, Caenorhabditis elegans , zygotes divide unequally because the first mitotic spindle is displaced off-center by the pull of stronger forces on its posterior end. Grill et al. investigated whether this asymmetry is caused by differences in strength, number, or spatial positioning of the force generators (presumably cortical motors) that pull on the microtubules. They determined the distribution of force generators in a living C. elegans embryo while the spindle was becoming displaced to one side of the embryo by destroying the centrosome and analyzing the movement of resulting aster fragments toward the cortex. They found that more force-generating elements are available for the posterior spindle pole. S. W. Grill, J. Howard, E. Schäffer, E. H. K. Stelzer, A. A. Hyman, The distribution of active force generators controls mitotic spindle position. Science 301 , 518-521 (2003). [Abstract] [Full Text]

Mechanisms of cell division: lessons from a nematode

The mechanisms orchestrating spatial cell division control remain poorly understood. In animal cells, the position of the mitotic spindle dictates cleavage furrow placement, and thus plays a key role in governing spatial relationships between resulting daughter cells. The one-cell stage Caenorhabditis elegans embryo is an attractive model system to investigate the mechanisms underlying spindle positioning in metazoans. In this review, the experimental advantages of this model system for an in vivo dissection of cell division processes are first discussed. Next, three lines of experiments that were conducted to dissect the mechanisms governing spindle positioning in one-cell stage C. elegans embryos are summarized. First, localized laser micro-irradiations were utilized to identify the forces acting on spindle poles during anaphase. This work revealed that there is a precise imbalance of pulling forces acting on the two spindle poles, with the forces acting on the posterior spindle pole being in slight excess, thus explaining the asymmetric spindle position achieved by the end of anaphase. Second, an RNAi-based functional genomic screen was carried out to identify novel components required for generating these pulling forces. This uncovered that gpr-1/gpr-2, which encode GoLoco-containing proteins, as well as the previously identified Ga subunits goa-1/gpa-16, are required for generation of pulling forces on the spindle poles. Third, the zyg-8 locus was identified by mutational analysis to play a distinct role during anaphase spindle positioning. zyg-8 was found to encode a protein related to human Doublecortin, which is affected in patients with neuronal migration disorders. Moreover, ZYG-8 is a microtubule-associated protein that stabilizes microtubules against depolymerization. Together, these experimental approaches contribute to a better understanding of the mechanisms orchestrating spatial cell division control in metazoan organisms.

Spindle positioning during the asymmetric first cell division of Caenorhabditis elegans embryos.

Cell division during development in many cases generates daughter cells that differ not only in fate, but also in size. We investigate the mechanisms that ensure proper spindle positioning during such asymmetric divisions using the one-cell stage Caenorhabditis elegans embryo as a model system. We utilized a UV laser microbeam as an in vivo microtubule-severing device to probe the forces driving spindle positioning. Our results indicate that extra-spindle pulling forces acting on the spindle poles dictate spindle position along the anterior-posterior embryonic axis. Importantly, forces acting on the posterior spindle pole appear more extensive than those acting on the anterior one, thus explaining the overall posterior spindle displacement that leads to the asymmetric division of the wild-type one-cell stage embryo. In separate work, we analysed a locus called zyg-8, which plays a key role in ensuring proper spindle positioning. Our data show that zyg-8 is required to promote microtubule growth and/or stability during anaphase. We identified the molecular nature of the zyg-8 locus in the course of a large-scale RNAi-based functional genomics screen. ZYG-8 harbours two notable protein domains: a Ca2+/calmodulin-dependent kinase domain, and a domain related to doublecortin, a human microtubule-associated protein involved in neuronal migration.

Posterior beta spindles evoked by photic stimulation

Posterior beta spindles evoked by photic stimulation

Study of cortical spindles during sleep in the rat

TERRIER, G. AND CL. GOTTESMANN. Study of cortical spindles during sleep in the rat. BRAIN RES. BULL. 3(6) 701–706, 1978.—A systematic analysis of the dorsally accessible cortical areas has been undertaken so as to study the space-time distribution, the intrinsic characteristics and the physiological appearance modalities of cortical spindles in the rat. After falling asleep, the animal presents anterior spindles of which the number, duration and amplitude increase as sleep with slow waves deepens. These spindles are fully developed during intermediate state which precedes and follows paradoxical sleep, where they are associated with a theta rhythm in the hippocampus and the visual cortex. These spindles, at their maximum in anterior regions, lose intensity as one approaches the cerebellum. Uniquely during slow wave phase of sleep, the rat presents posterior spindles which are predominant in the occipital region, are less marked in parietal and cerebellar regions and disappear in the frontal cortex. Less frequent than anterior spindles, they are significantly distinguished by their lower amplitude, higher frequency and shorter duration.