Bursting Properties Research Articles

Identification of molecular determinants of gene-specific bursting patterns by high-throughput imaging screens.

Stochastic transcriptional bursting is a universal property of active genes. While different genes exhibit distinct bursting patterns, the molecular mechanisms for gene-specific stochastic bursting are largely unknown. We have developed and applied a high-throughput-imaging based screening strategy to identify cellular factors and molecular mechanisms that determine the bursting behavior of human genes. Focusing on epigenetic regulators, we find that protein acetylation is a strong acute modulator of burst frequency, burst size and heterogeneity of bursting. Acetylation globally affects the Off-time of genes but has gene-specific effects on the On-time. Yet, these effects are not strongly linked to promoter acetylation, which do not correlate with bursting properties, and forced promoter acetylation has variable effects on bursting. Instead, we demonstrate acetylation of the Integrator complex as a key determinant of gene bursting. Specifically, we find that elevated Integrator acetylation decreases bursting frequency. Taken together our results suggest a prominent role of non-histone proteins in determining gene bursting properties, and they identify histone-independent acetylation of a transcription cofactor as an allosteric modulator of bursting via a far-downstream bursting checkpoint.

Bifurcations and bursting in the Epileptor.

The Epileptor is a phenomenological model for seizure activity that is used in a personalized large-scale brain modeling framework, the Virtual Epileptic Patient, with the aim of improving surgery outcomes for drug-resistant epileptic patients. Transitions between interictal and ictal states are modeled as bifurcations, enabling the definition of seizure classes in terms of onset/offset bifurcations. This establishes a taxonomy of seizures grounded in their essential underlying dynamics and the Epileptor replicates the activity of the most common class, as observed in patients with focal epilepsy, which is characterized by square-wave bursting properties. The Epileptor also encodes an additional mechanism to account for interictal spikes and spike and wave discharges. Here we use insights from a more generic model for square-wave bursting, based on the Unfolding Theory approach, to guide the bifurcation analysis of the Epileptor and gain a deeper understanding of the model and the role of its parameters. We show how the Epileptor's parameters can be modified to produce activities for other seizures classes of the taxonomy, as observed in patients, so that the large-scale brain models could be further personalized. Some of these classes have already been described in the literature in the Epileptor, others, predicted by the generic model, are new. Finally, we unveil how the interaction with the additional mechanism for spike and wave discharges alters the bifurcation structure of the main burster.

Age-dependent dysregulation of locus coeruleus firing in a transgenic rat model of Alzheimer's disease

Hyperphosphorylated tau in the locus coeruleus (LC) is ubiquitous in prodromal Alzheimer's disease (AD), and LC neurons degenerate as AD progresses. Hyperphosphorylated tau alters firing rates in other brain regions, but its effects on LC neurons are unknown. We assessed single unit LC activity in anesthetized wild-type (WT) and TgF344-AD rats at 6 months, which represents a prodromal stage when LC neurons are the only cells containing hyperphosphorylated tau in TgF344-AD animals, and at 15 months when amyloid-β (Aβ) and tau pathology are both abundant in the forebrain. At baseline, LC neurons from TgF344-AD rats were hypoactive at both ages compared to WT littermates but showed elevated spontaneous bursting properties. Differences in footshock-evoked LC firing depended on age, with 6-month TgF344-AD rats demonstrating aspects of hyperactivity, and 15-month transgenic rats showing hypoactivity. Early LC hyperactivity is consistent with appearance of prodromal neuropsychiatric symptoms and is followed by LC hypoactivity which contributes to cognitive impairment. These results support further investigation into disease stage-dependent noradrenergic interventions for AD.

Alzheimer’s Disease risk gene SORL1 affects synaptic function in hiPSC‐derived cortical neurons

AbstractBackgroundThe cellular endo‐lysosomal network (ELN) is implicated in Alzheimer’s Disease pathology and Sortilin‐related receptor 1 (SORL1) is important for normal function of the neuronal ELN. Loss of SORL1 in human induced pluripotent stem cell (hiPSC) derived neurons replicates early AD cytopathology, such as enlarged early endosomes. Work in our laboratory has shown that SORL1 deficiency results in decreased cell‐surface expression of glutamate receptor subunit GLUR1 and neurotrophic receptor TRKB. RNAseq of SORL1 deficient neurons revealed alterations in synaptic‐associated gene ontology transcripts. Together, these data implicate a potential functional consequence for SORL1 deletion. Here we investigate the impact of SORL1 on hiPSC‐derived neuronal synaptic function.MethodNeuronal cultures were differentiated using dual SMAD inhibition of previously generated wild‐type and SORL1 deficient hiPSCs. Following differentiation, neuronal cultures were either purified using cell surface markers CD44, CD184, and CD271 or passaged as mixed cultures for immunocytochemical (ICC) or electrophysiological experiments, respectively. For ICC, neurons were then fixed with paraformaldehyde and stained with experiment‐appropriate antibodies before imaging on a confocal scanning microscope. Mixed cultures were plated onto Axion Biosystems multiple electrode array (MEA) culture plates for electrophysiological analysis. MEA cultures were recorded twice a week for up to ten weeks post‐plating. Recordings and analysis were performed using Axion Biosystems Maestro Pro system.ResultUsing ICC, we find alterations in alterations in synaptic protein and glutamate receptor co‐localization in SORL1 deficient neuronal cultures. When cultured on MEA plates, wild‐type and SORL1 deficient neuronal cultures are active and robust. Neuronal cultures develop glutamate signaling dependent patterns of activity, or network bursting. Neural firing rates are increased in SORL1 deficient neurons. Network bursting properties in SORL1 deficient are altered in comparison to wild‐type cultures, consistent with increased neural excitability.ConclusionLoss of SORL1 alters hiPSC‐derived neuronal activity and synapse‐dependent activity. Further investigation is warranted into the alterations in neuronal physiology and will help us understand the role AD‐related ELN disruption plays in synaptic pathologies.

Estrogens rapidly shape synaptic and intrinsic properties to regulate the temporal precision of songbird auditory neurons.

Sensory neurons parse millisecond-variant sound streams like birdsong and speech with exquisite precision. The auditory pallial cortex of vocal learners like humans and songbirds contains an unconventional neuromodulatory system: neuronal expression of the estrogen synthesis enzyme aromatase. Local forebrain neuroestrogens fluctuate when songbirds hear a song, and subsequently modulate bursting, gain, and temporal coding properties of auditory neurons. However, the way neuroestrogens shape intrinsic and synaptic properties of sensory neurons remains unknown. Here, using a combination of whole-cell patch clamp electrophysiology and calcium imaging, we investigate estrogenic neuromodulation of auditory neurons in a region resembling mammalian auditory association cortex. We found that estradiol rapidly enhances the temporal precision of neuronal firing via a membrane-bound G-protein coupled receptor and that estradiol rapidly suppresses inhibitory synaptic currents while sparing excitation. Notably, the rapid suppression of intrinsic excitability by estradiol was predicted by membrane input resistance and was observed in both males and females. These findings were corroborated by analysis of in vivo electrophysiology recordings, in which local estrogen synthesis blockade caused acute disruption of the temporal correlation of song-evoked firing patterns. Therefore, on a modulatory timescale, neuroestrogens alter intrinsic cellular properties and inhibitory neurotransmitter release to regulate the temporal precision of higher-order sensory neurons.

Asymmetric and transient properties of reciprocal activity of antagonists during the paw-shake response in the cat.

Mutually inhibitory populations of neurons, half-center oscillators (HCOs), are commonly involved in the dynamics of the central pattern generators (CPGs) driving various rhythmic movements. Previously, we developed a multifunctional, multistable symmetric HCO model which produced slow locomotor-like and fast paw-shake-like activity patterns. Here, we describe asymmetric features of paw-shake responses in a symmetric HCO model and test these predictions experimentally. We considered bursting properties of the two model half-centers during transient paw-shake-like responses to short perturbations during locomotor-like activity. We found that when a current pulse was applied during the spiking phase of one half-center, let’s call it #1, the consecutive burst durations (BDs) of that half-center increased throughout the paw-shake response, while BDs of the other half-center, let’s call it #2, only changed slightly. In contrast, the consecutive interburst intervals (IBIs) of half-center #1 changed little, while IBIs of half-center #2 increased. We demonstrated that this asymmetry between the half-centers depends on the phase of the locomotor-like rhythm at which the perturbation was applied. We suggest that the fast transient response reflects functional asymmetries of slow processes that underly the locomotor-like pattern; e.g., asymmetric levels of inactivation across the two half-centers for a slowly inactivating inward current. We compared model results with those of in-vivo paw-shake responses evoked in locomoting cats and found similar asymmetries. Electromyographic (EMG) BDs of anterior hindlimb muscles with flexor-related activity increased in consecutive paw-shake cycles, while BD of posterior muscles with extensor-related activity did not change, and vice versa for IBIs of anterior flexors and posterior extensors. We conclude that EMG activity patterns during paw-shaking are consistent with the proposed mechanism producing transient paw-shake-like bursting patterns found in our multistable HCO model. We suggest that the described asymmetry of paw-shaking responses could implicate a multifunctional CPG controlling both locomotion and paw-shaking.

The effect of green biobased binder on structural, mechanical, liquid absorption and wetting properties of coated papers

Synthetic styrene-butadiene (SB) and styrene-acrylic (SA) latex binders used in paper coating formulations are common and based on unsustainable petroleum sources. Today's papermaking industry turns towards sustainable substitutes that do not compromise quality, and reduce carbon emission, toxic substance release and waste disposal concerns related to fossil fuel sources. In this study, colloidal starch-based latex nanoparticles that do not require cooking were used for pigment coating and coated on the paper surfaces. The effects of these new biobased binders on the structural and mechanical strength properties, liquid absorption, wetting and surface topography of the paper were investigated and compared with the properties of coated paper that SB latex was used as a coating binder. The results demonstrated that the biobased latex decreases the grammage of paper while maintaining an identical thickness relative to the SB latex samples. The porosity, permeability and roughness of biobased latex are found higher than the SB latex. The biobased latex successfully replaced SB latex at 1:1 ratio and enabled an equivalent of bursting, tear, tensile strength and structural properties. Synopsis : A plant-based starch nanoparticle latex is engineered to replace petroleum based synthetic latexes without compromising structural and mechanical paper properties.

Intrinsic and synaptic mechanisms controlling the expiratory activity of excitatory lateral parafacial neurones of rats.

Active expiration is essential for increasing pulmonary ventilation during high chemical drive (hypercapnia). The lateral parafacial (pFL ) region, which contains expiratory neurones, drives abdominal muscles during active expiration in response to hypercapnia. However, the electrophysiological properties and synaptic mechanisms determining the activity of pFL expiratory neurones, as well as the specific conditions for their emergence, are not fully understood. Using whole cell electrophysiology and single cell quantitative RT-PCR techniques, we describe the intrinsic electrophysiological properties, the phenotype and the respiratory-related synaptic inputs to the pFL expiratory neurones, as well as the mechanisms for the expression of their expiratory activity under conditions of hypercapnia-induced active expiration, using in situ preparations of juvenile rats. We also evaluated whether these neurones possess intrinsic CO2 /[H+ ] sensitivity and burst generating properties. GABAergic and glycinergic inhibition during inspiration and expiration suppressed the activity of glutamatergic pFL expiratory neurones in normocapnia. In hypercapnia, these neurones escape glycinergic inhibition and generate burst discharges at the end of expiration. Evidence for the contribution of post-inhibitory rebound, CaV 3.2 isoform of T-type Ca2+ channels and intracellular [Ca2+ ] is presented. Neither intrinsic bursting properties, mediated by persistent Na+ current, nor CO2 /[H+ ] sensitivity or expression of CO2 /[H+ ] sensitive ion channels/receptors (TASK or GPR4) were observed. On the other hand, hyperpolarisation-activated cyclic nucleotide-gated and twik-related K+ leak channels were recorded. Post-synaptic disinhibition and the intrinsic electrophysiological properties of glutamatergic neurones play important roles in the generation of the expiratory oscillations in the pFL region during hypercapnia in rats. KEY POINTS: Hypercapnia induces active expiration in rats and the recruitment of a specific population of expiratory neurones in the lateral parafacial (pFL ) region. Post-synaptic GABAergic and glycinergic inhibition both suppress the activity of glutamatergic pFL neurones during inspiratory and expiratory phases in normocapnia. Hypercapnia reduces glycinergic inhibition during expiration leading to burst generation by pFL neurones; evidence for a contribution of post-inhibitory rebound, voltage-gated Ca2+ channels and intracellular [Ca2+ ] is presented. pFL glutamatergic expiratory neurones are neither intrinsic burster neurones, nor CO2 /[H+ ] sensors, and do not express CO2 /[H+ ] sensitive ion channels or receptors. Post-synaptic disinhibition and the intrinsic electrophysiological properties of glutamatergic neurones both play important roles in the generation of the expiratory oscillations in the pFL region during hypercapnia in rats.

Burst-induced coronal cooling in GS 1826–24

Type I X-ray bursts in GS 1826–24, and in several other systems, may induce cooling of the hot inner accretion flow that surrounds the bursting neutron star. Given that GS 1826–24 remained persistently in the hard state over the period 2003–2008 and presented regular bursting properties, we stacked the spectra of the X-ray bursts detected by INTEGRAL (JEM-X and ISGRI) and XMM-Newton (RGS) during that period to study the effect of the burst photons on the properties of the Comptonizing medium. The extended energy range provided by these instruments allows the simultaneous observation of the burst and persistent emission spectra. We detect an overall change in the shape of the persistent emission spectrum in response to the burst photon shower. For the first time, we observe simultaneously a drop in the hard X-ray emission, together with a soft X-ray excess with respect to the burst blackbody emission. The hard X-ray drop can be explained by burst-induced coronal cooling, while the bulk of the soft X-ray excess can be described by fitting the burst emission with an atmosphere model, instead of a simple blackbody model. Traditionally, the persistent emission was assumed to be invariant during X-ray bursts, and more recently to change only in normalization but not in spectral shape; the observed change in the persistent emission level during X-ray bursts may thus trigger the revision of existing neutron star mass-radius constraints, as the derived values rely on the assumption that the persistent emission does not change during X-ray bursts. The traditional burst fitting technique leads to up to a 10% overestimation of the bolometric burst flux in GS 1826–24, which significantly hampers the comparisons of the KEPLER and MESA model against this “textbook burster”.

The effect of fabric structure on the bursting characteristics of warp-knitted surgical mesh

Surgical mesh is a porous fabric that is mainly produced from polymeric monofilaments using the warp-knitting method. Surgical mesh is used for healing hernia. Among the mechanical properties of the mesh, tensile and bursting strength have special significance. In this research, warp-knitted surgical meshes with five structures have been produced using polypropylene monofilament, and the effect of the fabric structure on the bursting characteristics of them has been investigated. All the meshes exhibited supraphysiologic bursting strength. quasi-Sandfly mesh demonstrated the highest, while quasi-Marqussite mesh exhibited the least bursting characteristics. The increase in the bursting characteristics of the meshes is attributed to the increase in the areal density and decrease in the porosity of them. Moreover, conformity between the bursting and tensile properties of them in bias direction has been observed. Overall, quasi-Marqussite mesh was proposed as the most desirable mesh in terms of porosity, weight, and bursting characteristics.

Conductive fabrics made of polypropylene/multi-walled carbon nanotube coated polyester yarns: Mechanical properties and electromagnetic interference shielding effectiveness

Polypropylene (PP)/multi-walled carbon nanotube (MWCNT) coated polyethylene terephthalate (PET) yarns are made into conductive woven/knitted fabrics. The fabric structure, tensile properties, static bursting properties, electrical resistivity, and electromagnetic interference shielding effectiveness (EMI SE) of the conductive fabrics are evaluated in terms of the content of MWCNT. The test results indicate that the conductive fabrics have optimal tensile strength and optimal bursting strength when composed of 2 wt% MWCNT. Moreover, the conductive fabrics have a stabilized structure as well as the lowest electrical resistivity and an optimal EMI SE when composed of 8 wt% MWCNT. Similarly, 3-layered conductive fabrics also have the maximum EMI SE. The conductive fabrics produced in this study have permanent electrical properties, a light weight, and flexibility. Moreover, their EMI SE reaches the protection level required by standard electronic devices.

The central pattern generator underlying swimming in Dendronotus iris: a simple half-center network oscillator with a twist.

The nudibranch mollusc, Dendronotus iris, swims by rhythmically flexing its body from left to right. We identified a bilaterally represented interneuron, Si3, that provides strong excitatory drive to the previously identified Si2, forming a half-center oscillator, which functions as the central pattern generator (CPG) underlying swimming. As with Si2, Si3 inhibited its contralateral counterpart and exhibited rhythmic bursts in left-right alternation during the swim motor pattern. Si3 burst almost synchronously with the contralateral Si2 and was coactive with the efferent impulse activity in the contralateral body wall nerve. Perturbation of bursting in either Si3 or Si2 by current injection halted or phase-shifted the swim motor pattern, suggesting that they are both critical CPG members. Neither Si2 nor Si3 exhibited endogenous bursting properties when activated alone; activation of all four neurons was necessary to initiate and maintain the swim motor pattern. Si3 made a strong excitatory synapse onto the contralateral Si2 to which it is also electrically coupled. When Si3 was firing tonically but not exhibiting bursting, artificial enhancement of the Si3-to-Si2 synapse using dynamic clamp caused all four neurons to burst. In contrast, negation of the Si3-to-Si2 synapse by dynamic clamp blocked ongoing swim motor patterns. Together, these results suggest that the Dendronotus swim CPG is organized as a "twisted" half-center oscillator in which each "half" is composed of two excitatory-coupled neurons from both sides of the brain, each of which inhibits its contralateral counterpart. Consisting of only four neurons, this is perhaps the simplest known network oscillator for locomotion.

Development of pacemaker properties and rhythmogenic mechanisms in the mouse embryonic respiratory network.

Breathing is a vital rhythmic behavior generated by hindbrain neuronal circuitry, including the preBötzinger complex network (preBötC) that controls inspiration. The emergence of preBötC network activity during prenatal development has been described, but little is known regarding inspiratory neurons expressing pacemaker properties at embryonic stages. Here, we combined calcium imaging and electrophysiological recordings in mouse embryo brainstem slices together with computational modeling to reveal the existence of heterogeneous pacemaker oscillatory properties relying on distinct combinations of burst-generating INaP and ICAN conductances. The respective proportion of the different inspiratory pacemaker subtypes changes during prenatal development. Concomitantly, network rhythmogenesis switches from a purely INaP/ICAN-dependent mechanism at E16.5 to a combined pacemaker/network-driven process at E18.5. Our results provide the first description of pacemaker bursting properties in embryonic preBötC neurons and indicate that network rhythmogenesis undergoes important changes during prenatal development through alterations in both circuit properties and the biophysical characteristics of pacemaker neurons.

Network burst activity in hippocampal neuronal cultures: the role of synaptic and intrinsic currents.

The goal of this work was to define the contributions of intrinsic and synaptic mechanisms toward spontaneous network-wide bursting activity, observed in dissociated rat hippocampal cell cultures. This network behavior is typically characterized by short-duration bursts, separated by order of magnitude longer interburst intervals. We hypothesize that while short-timescale synaptic processes modulate spectro-temporal intraburst properties and network-wide burst propagation, much longer timescales of intrinsic membrane properties such as persistent sodium (Nap) currents govern burst onset during interburst intervals. To test this, we used synaptic receptor antagonists picrotoxin, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), and 3-(2-carboxypiperazine-4-yl)propyl-1-phosphonate (CPP) to selectively block GABAA, AMPA, and NMDA receptors and riluzole to selectively block Nap channels. We systematically compared intracellular activity (recorded with patch clamp) and network activity (recorded with multielectrode arrays) in eight different synaptic connectivity conditions: GABAA + NMDA + AMPA, NMDA + AMPA, GABAA + AMPA, GABAA + NMDA, AMPA, NMDA, GABAA, and all receptors blocked. Furthermore, we used mixed-effects modeling to quantify the aforementioned independent and interactive synaptic receptor contributions toward spectro-temporal burst properties including intraburst spike rate, burst activity index, burst duration, power in the local field potential, network connectivity, and transmission delays. We found that blocking intrinsic Nap currents completely abolished bursting activity, demonstrating their critical role in burst onset within the network. On the other hand, blocking different combinations of synaptic receptors revealed that spectro-temporal burst properties are uniquely associated with synaptic functionality and that excitatory connectivity is necessary for the presence of network-wide bursting. In addition to confirming the critical contribution of direct excitatory effects, mixed-effects modeling also revealed distinct combined (nonlinear) contributions of excitatory and inhibitory synaptic activity to network bursting properties.

Modeling thalamic dynamics with a network of integrate and fire neurons

Spindle oscillations are rhythmic oscillations at 8-14 Hz originated in the thalamus during slow-wave sleep. The generation of this rhythm is due to the rebound bursting properties of thalamo-cortical (TC) relay cells, which are mutually connected with the thalamic reticular neurons (RE). Several experiments have shown that during sleep TC neurons can produce bursts of action potentials due to disinhibition from RE neurons. These bursts in turn trigger an inhibitory rebound in TC neurons thus giving rise to an endogenous oscillatory activity [1,2]. Various sophisticated HH-based models have described in detail the electrophysiological properties of these neurons and replicate most of the experimental observations made in-vivo and in-vitro [2]. However these types of models are computationally very expensive, thus raising the question of whether this dynamical behavior can be reproduced in simpler models. To address this question, we used an adaptive exponential integrate and fire model (aEIF) [3] and studied the conditions under which spindle oscillations appear in large neuronal networks. The aEIF model can reproduce the rebound bursting properties of thalamic cells and has been reported to replicate the self-sustained oscillatory behavior in a minimal 4-neuron circuit [4]. In this minimal motif, RE cells exhibit periodic sequences of bursts at 10 Hz (spindles) and recruit TC neurons, which oscillate in anti-phase at half the frequency (cycle skipping) tightly correlated with IPSP bursts. However, it is unclear whether these spindle oscillations are preserved when the neurons are interconnected in a larger network, while keeping their intrinsic properties. In order to examine this issue, we have systematically extended the size of the network of TC and RE neurons, and checked whether the features of the above-described oscillatory activity hold out. We found the reciprocal interaction between TC and RE to be essential for sustaining this activity, coherently with experimental results [5]. Furthermore, some of the RE neurons exhibit periodic bursting at the spindle frequency, provided they are not connected to TC neurons directly, but only indirectly through local inhibitory connections with RE neighbors. Interestingly, this behavior seems to require that the RE layer has a certain degree of clustering. Under these conditions, a few TC neurons in the network respond with a burst every two cycles (cycle skipping). The fact that most TC do not exhibit cycle skipping could associated with a large strength of GABA IPSPs impinging on them, according to results of HH kinetic models [6]. In summary, our aEIF network model provides a computationally efficient description of the dynamical features of the thalamus while preserving the properties of the individual neurons.

Observation of rock fragment ejection in post-failure response

Rockburst is a dynamic energy phenomenon accompanying the failure of hard rock in the form of brittle fractures [1–3]. Surplus energy is released when the accumulated elastic strain energy of rock is greater than the consumptive energy required for fracturing during its quick post-failure. This freed energy often takes radiant forms, such as acoustic energy, electromagnetic energy, and thermal energy [2,4,5], as well as physical forms, such as fragments being thrown in the laboratory, or an ejective rockburst in the field [6–11]. Of these types of releases, the fragments with kinetic energy always result in dangerous engineering accidents due to their abrupt impacts at high speeds [12–17]. To reduce bursting disasters in underground engineering, a great deal of effort has been put forth to explain the rockburst mechanism, predict ejection behavior, and design corresponding protective support techniques, etc. [18–24]. Conventional wisdom indicated that knowledge of the ejection velocity of the burst rock fragments is the key to further predicting rockburst risks and designing a corresponding protective support system [25–28]. For example, Kaiser and Cai underlined that rock ejection with a velocity of up to 3 m/s required a specially integrated yielding support system [28]. Simulated rockburst experiments in an underground tunnel in South Africa were successfully conducted to measure the ejection velocity of rock fragments directly. The results indicated that the in situ rock fragments were ejected from the tunnel wall with velocities in the range of 0.6 m/s to 2.5 m/s, thereby providing useful clues for rockburst's prevention technique in deep mining [29,30]. As expected, high experimental costs, rigorous site requirements (i.e., hard rock and high geostress) and complicated preparatory work for such field experiments have limited the application of in-situ rockburst measurements. Laboratory observations of rock ejections serve as a convenient and reasonable alternative method [31–33]. Petukhov noted that violent fracture of rock specimen in a compression machine could represent a laboratory simulation for dynamic rock failure during a burst in the field [32,34]. Visual observations of fragment ejection velocity in a laboratory are important not only to understand further the outbreak behavior of fragments in post-failure of hard rock but also to assess the bursting properties of rock in actual engineering. High-speed filming, which records instantaneous behavior by taking successive photographs over a short time interval, has been used in dynamically mechanical experiments as an accurate tool for observing impact failure [33,35,36]. Unfortunately, the highspeed camera has few applications in the documentation of ejection performance rock specimens under quasi-steady compression conditions. As a result, these filming methods warrant further research. In this article, we present a method to observe the ejection velocity of rock fragments under laboratory uniaxial compression tests with the help of a high-speed camera. Also shown in this article is an algorithmic program developed to calculate the initial speed and throwing angle of ejected rock fragments. This work

Components of action potential repolarization in cerebellar parallel fibres.

Repolarization of the presynaptic action potential is essential for transmitter release, excitability and energy expenditure. Little is known about repolarization in thin, unmyelinated axons forming en passant synapses, which represent the most common type of axons in the mammalian brain's grey matter.We used rat cerebellar parallel fibres, an example of typical grey matter axons, to investigate the effects of K(+) channel blockers on repolarization. We show that repolarization is composed of a fast tetraethylammonium (TEA)-sensitive component, determining the width and amplitude of the spike, and a slow margatoxin (MgTX)-sensitive depolarized after-potential (DAP). These two components could be recorded at the granule cell soma as antidromic action potentials and from the axons with a newly developed miniaturized grease-gap method. A considerable proportion of fast repolarization remained in the presence of TEA, MgTX, or both. This residual was abolished by the addition of quinine. The importance of proper control of fast repolarization was demonstrated by somatic recordings of antidromic action potentials. In these experiments, the relatively broad K(+) channel blocker 4-aminopyridine reduced the fast repolarization, resulting in bursts of action potentials forming on top of the DAP. We conclude that repolarization of the action potential in parallel fibres is supported by at least three groups of K(+) channels. Differences in their temporal profiles allow relatively independent control of the spike and the DAP, whereas overlap of their temporal profiles provides robust control of axonal bursting properties.

Respiratory neuron characterization reveals intrinsic bursting properties in isolated adult turtle brainstems (Trachemys scripta)

It is not known whether respiratory neurons with intrinsic bursting properties exist within ectothermic vertebrate respiratory control systems. Thus, isolated adult turtle brainstems spontaneously producing respiratory motor output were used to identify and classify respiratory neurons based on their firing pattern relative to hypoglossal (XII) nerve activity. Most respiratory neurons (183/212) had peak activity during the expiratory phase, while inspiratory, post-inspiratory, and novel pre-expiratory neurons were less common. During synaptic blockade conditions, ∼10% of respiratory neurons fired bursts of action potentials, with post-inspiratory cells (6/9) having the highest percentage of intrinsic burst properties. Most intrinsically bursting respiratory neurons were clustered at the level of the vagus (X) nerve root. Synaptic inhibition blockade caused seizure-like activity throughout the turtle brainstem, which shows that the turtle respiratory control system is not transformed into a network driven by intrinsically bursting respiratory neurons. We hypothesize that intrinsically bursting respiratory neurons are evolutionarily conserved and represent a potential rhythmogenic mechanism contributing to respiration in adult turtles.

Prostaglandin E2 differentially modulates the central control of eupnoea, sighs and gasping in mice.

Prostaglandin E2 (PGE2) augments distinct inspiratory motor patterns, generated within the preBötzinger complex (preBötC), in a dose-dependent way. The frequency of sighs and gasping are stimulated at low concentrations, while the frequency of eupnoea increases only at high concentrations. We used in vivo microinjections into the preBötC and in vitro isolated brainstem slice preparations to investigate the dose-dependent effects of PGE2 on the preBötC activity. Synaptic measurements in whole cell voltage clamp recordings of inspiratory neurons revealed no changes in inhibitory or excitatory synaptic transmission in response to PGE2 exposure. In current clamp recordings obtained from inspiratory neurons of the preBötC, we found an increase in the frequency and amplitude of bursting activity in neurons with intrinsic bursting properties after exposure to PGE2. Riluzole, a blocker of the persistent sodium current, abolished the effect of PGE2 on sigh activity, while flufenamic acid, a blocker of the calcium-activated non-selective cation conductance, abolished the effect on eupnoeic activity caused by PGE2. Prostaglandins are important regulators of autonomic functions in the mammalian organism. Here we demonstrate in vivo that prostaglandin E2 (PGE2) can differentially increase the frequency of eupnoea (normal breathing) and sighs (augmented breaths) when injected into the preBötzinger complex (preBötC), a medullary area that is critical for breathing. Low concentrations of PGE2 (100-300 nm) increased the sigh frequency, while higher concentrations (1-2 μm) were required to increase the eupnoeic frequency. The concentration-dependent effects were similarly observed in the isolated preBötC. This in vitro preparation also revealed that riluzole, a blocker of the persistent sodium current (INap), abolished the modulatory effect on sighs, while flufenamic acid, an antagonist for the calcium-activated non-selective cation conductance (ICAN ) abolished the effect of PGE2 on fictive eupnoea at higher concentrations. At the cellular level PGE2 significantly increased the amplitude and frequency of intrinsic bursting in inspiratory neurons. By contrast PGE2 affected neither excitatory nor inhibitory synaptic transmission. We conclude that PGE2 differentially modulates sigh, gasping and eupnoeic activity by differentially increasing INap and ICAN currents in preBötC neurons.

NuSTAR DISCOVERY OF A 3.76 s TRANSIENT MAGNETAR NEAR SAGITTARIUS A*

We report the discovery of 3.76-s pulsations from a new burst source near Sgr A* observed by the NuSTAR Observatory. The strong signal from SGR J1745-29 presents a complex pulse profile modulated with pulsed fraction 27+/-3 % in the 3-10 keV band. Two observations spaced 9 days apart yield a spin-down rate of Pdot = (6.5+/-1.4)x10^{-12}. This implies a magnetic field B = 1.6x10^14 G, spin-down power Edot = 5x10^33 erg/s, and characteristic age P/2Pdot = 9x10^3 yr, for the rotating dipole model. However, the current Pdot may be erratic, especially during outburst. The flux and modulation remained steady during the observations and the 3-79 keV spectrum is well fitted by a combined blackbody plus power-law model with temperature kT_BB = 0.96+/-0.02 keV and photon index = 1.5+/-0.4, respectively. The neutral hydrogen column density (nH ~ 1.4x10^23 cm^{-2}) measured by NuSTAR and Swift suggests that SGR J1745-29 is located at or near the Galactic Center. The lack of an X-ray counterpart in the published Chandra survey catalog sets a quiescent 2-8 keV luminosity limit of Lx ~< 10^32 erg/s. The bursting, timing, and spectral properties indicate a transient magnetar undergoing an outburst with 2-79 keV luminosity up to 3.5x10^35 erg/s for a distance of 8 kpc. SGR J1745-29 joins a growing subclass of transient magnetars, indicating that many magnetars in quiescence remain undetected in the X-ray band or have been detected as high-B radio pulsars. The peculiar location of SGR J1745-29 has important implications for the formation and dynamics of neutron stars in the Galactic Center region.