Mechanism Of Potentiation Research Articles

Piperine enhances contractile force in slow- and fast-twitch muscle.

Piperine has been shown to bind to myosin and shift the distribution of conformational states of myosin molecules from the super-relaxed state to the disordered relaxed state. However, little is known about the implications for muscle force production and potential underlying mechanisms. Muscle contractility experiments were performed using isolated muscles and single fibres from rats and mice. The dose-response effect of piperine on muscle force was assessed at several stimulation frequencies. The potentiation of muscle force was also tested in muscles fatigued by eccentric contractions. Potential mechanisms of force potentiation were assessed by measuring Ca2+ levels during stimulation in enzymatically dissociated muscle fibres, while myofibrillar Ca2+ sensitivity was assessed in chemically skinned muscle fibres. Piperine caused a dose-dependent increase in low-frequency force with no effect on high-frequency force in both slow- and fast-twitch muscle, with similar relative increases in twitch force, rate of force development and relaxation rate. The potentiating effect of piperine on low-frequency force was reversible, and piperine partially recovered low-frequency force in fatigued muscle. Piperine had no effect on myoplasmic free [Ca2+] levels in mouse muscle fibres, whereas piperine substantially augmented the force response to submaximal levels of [Ca2+] in rat MyHCII fibres and MyHCI fibres along with a minor increase in maximum Ca2+-activated force. Piperine enhances low-frequency force production in both fast- and slow-twitch muscle. The effects are reversible and can counteract muscle fatigue. The primary underlying mechanism appears to be an increase in Ca2+ sensitivity. KEY POINTS: Piperine is a plant alkaloid derived from black pepper. It is known to bind to skeletal muscle myosin and enhance resting ATP turnover but its effects on contractility are not well known. We showed for the first time a piperine-induced force potentiation that was pronounced during low-frequency electrical stimulation of isolated muscles. The effect of piperine was observed in both slow and fast muscle types, was reversible, and could counteract the force decrements observed after fatiguing muscle contractions. Piperine treatment caused an increase in myofibrillar Ca2+ sensitivity in chemically skinned muscle fibres, while we observed no effect on intracellular Ca2+ concentrations during electrical stimulation in enzymatically dissociated muscle fibres.

Effects of Cholinergic Receptor Activation and Magnetic Fields on Motor Behavior in Ischemic Gerbils

Introduction: Ischemic stroke stands as a leading global cause of death and disability, prompting the need for animal model experiments in stroke research and the protection of motor function. Recently, magnetic fields have gained significant interest in various biological contexts, showing promise in preserving neurons and reversing behavioral and morphological changes in stroke models. This study explores the potential synergy between static magnetic field and nAChR agonist administration in safeguarding motor behavior in ischemic gerbils. Objective: To determine whether the combined use of a static magnetic field and an agonist for nicotinic acetylcholine receptors (nAChR) can preserve motor behavior in ischemic gerbils.Methods: In this experimental study, 72 Mongolian gerbils were randomly allocated into nine groups (n=8): S, SISM, SINSM, ISM, INP, ISP, INSM, INNP, INSP, distributed according to surgical procedure and treatment. The animals were trained and evaluated on the Rotarod (RR) to assess motor performance.Results: The main finding was the preservation of motor behavior in the Sham Ischemia and Nicotine and Sham Magnetic Stimulation (SINSM) and Ischemia and Nicotine and South Pole Magnetic Field (INSP) groups, as evidenced by the results of the RR test.Conclusions: The findings are consistent with previous literature and provide insight into the mechanism of potentiation, as results showed that adding a nAChR agonist to the magnetic field preserved motor performance in the RR test of ischemic animals.

Structural mechanisms of α7 nicotinic receptor allosteric modulation and activation

The α7 nicotinic acetylcholine receptor is a pentameric ligand-gated ion channel that plays an important role in cholinergic signaling throughout the nervous system. Its unique physiological characteristics and implications in neurological disorders and inflammation make it a promising but challenging therapeutic target. Positive allosteric modulators overcome limitations of traditional α7 agonists, but their potentiation mechanisms remain unclear. Here, we present high-resolution structures of α7-modulator complexes, revealing partially overlapping binding sites but varying conformational states. Structure-guided functional and computational tests suggest that differences in modulator activity arise from the stable rotation of a channel gating residue out of the pore. We extend the study using a time-resolved cryoelectron microscopy (cryo-EM) approach to reveal asymmetric state transitions for this homomeric channel and also find that a modulator with allosteric agonist activity exploits a distinct channel-gating mechanism. These results define mechanisms of α7 allosteric modulation and activation with implications across the pentameric receptor superfamily.

Adenosine A2A receptors control generalization of contextual fear in rats

Fear learning is essential to survival, but traumatic events may lead to abnormal fear consolidation and overgeneralization, triggering fear responses in safe environments, as occurs in post-traumatic stress disorder (PTSD). Adenosine A2A receptors (A2AR) control emotional memory and fear conditioning, but it is not known if they affect the consolidation and generalization of fear, which was now investigated. We now report that A2AR blockade through systemic administration of the A2AR antagonist SCH58261 immediately after contextual fear conditioning (within the consolidation window), accelerated fear generalization. Conversely, A2AR activation with CGS21680 decreased fear generalization. Ex vivo electrophysiological recordings of field excitatory post-synaptic potentials (fEPSPs) in CA3-CA1 synapses and of population spikes in the lateral amygdala (LA), showed that the effect of SCH58261 is associated with a reversion of fear conditioning-induced decrease of long-term potentiation (LTP) in the dorsal hippocampus (DH) and with increased amplitude of LA LTP in conditioned animals. These data suggest that A2AR are engaged during contextual fear consolidation, controlling long-term potentiation mechanisms in both DH and LA during fear consolidation, impacting on fear generalization; this supports targeting A2AR during fear consolidation to control aberrant fear processing in PTSD and other fear-related disorders.

Predicting molecular mechanism of silymarin-potentiated diclofenac toxicity: Insight from in silico molecular docking

Silymarin was shown to enhance diclofenac toxicity by inducing the loss of mitochondrial membrane permeability (MMP) in Caco-2 cells, independent of endoplasmic reticulum stress. This study employed in silico molecular docking to further investigate the potential interaction between silymarin and specific mitochondrial proteins involved in the loss of mitochondria integrity, aiming to elucidate the underlying mechanism of potentiation. The target proteins for our docking analysis included mitochondrial complex I and III, voltage-dependent anion-selective channel (VDAC), and cyclophilin D (CypD). Our results indicated that diclofenac could bind to both mitochondrial complex I and III. In contrast, silymarin exhibited a strong interaction with mitochondrial complex I with the binding energy (ΔG) −7.74 kcal/mol and the inhibition constant (Ki) 2.12 µM, while not showing significant interaction with mitochondrial complex III. Additionally, silymarin had the potential to induce the opening of mitochondrial permeability transition pore by binding with VDAC in the outer mitochondrial membrane with ΔG −6.08 kcal/mol and Ki 34.94 µM. However, silymarin did not exhibit significant interaction with CypD in the inner mitochondrial membrane. Therefore, mitochondrial complex I and VDAC could be the potentiation targets of silymarin, resulting in the disruption of mitochondria integrity and enhancing the toxicity of diclofenac.

Presynaptic Kainate Receptors onto Somatostatin Interneurons Are Recruited by Activity throughout Development and Contribute to Cortical Sensory Adaptation.

Somatostatin (SST) interneurons produce delayed inhibition because of the short-term facilitation of their excitatory inputs created by the expression of metabotropic glutamate receptor 7 (mGluR7) and presynaptic GluK2-containing kainate receptors (GluK2-KARs). Using mice of both sexes, we find that as synaptic facilitation at layer (L)2/3 SST cell inputs increases during the first few postnatal weeks, so does GluK2-KAR expression. Removal of sensory input by whisker trimming does not affect mGluR7 but prevents the emergence of presynaptic GluK2-KARs, which can be restored by allowing whisker regrowth or by acute calmodulin activation. Conversely, late trimming or acute inhibition of Ca2+/calmodulin-dependent protein kinase II is sufficient to reduce GluK2-KAR activity. This developmental and activity-dependent regulation also produces a specific reduction of L4 GluK2-KARs that advances in parallel with the maturation of sensory processing in L2/3. Finally, we find that removal of both GluK2-KARs and mGluR7 from the synapse eliminates short-term facilitation and reduces sensory adaptation to repetitive stimuli, first in L4 of somatosensory cortex, then later in development in L2/3. The dynamic regulation of presynaptic GluK2-KARs potentially allows for flexible scaling of late inhibition and sensory adaptation.SIGNIFICANCE STATEMENT Excitatory synapses onto somatostatin (SST) interneurons express presynaptic, calcium-permeable kainate receptors containing the GluK2 subunit (GluK2-KARs), activated by high-frequency activity. In this study we find that their presence on L2/3 SST synapses in the barrel cortex is not based on a hardwired genetic program but instead is regulated by sensory activity, in contrast to that of mGluR7. Thus, in addition to standard synaptic potentiation and depression mechanisms, excitatory synapses onto SST neurons undergo an activity-dependent presynaptic modulation that uses GluK2-KARs. Further, we present evidence that loss of the frequency-dependent synaptic components (both GluK2-KARs and mGluR7 via Elfn1 deletion) contributes to a decrease in the sensory adaptation commonly seen on repetitive stimulus presentation.

A computational model of current control mechanism for long-term potentiation (LTP) in human episodic memory based on gene-gene interaction.

The establishment of long-term potentiation (LTP) is a prime process for the formation of episodic memory. During the establishment of LTP, activations of various components are required in the signaling cascade of the LTP pathway. Past efforts to determine the activation of components relied extensively on the cellular or molecular level. In this paper, we have proposed a computational model based on gene-level cascading and interaction in LTP signaling for the establishment and control of current signals for achieving the desired level of activation in the formation of episodic memory. This paper also introduces a model for a generalized signaling pathway in episodic memory. A back-propagation feedback mechanism is used for updating the interaction levels in the signaling cascade starting from the last stage and ending at the start stage of the signaling cascade. Simulation of the proposed model has been performed for the LTP signaling pathway in the context of human episodic memory. We found through simulation that the qualifying genes correction factors of all stages are updated to their maximum limit. The article explains the signaling pathway for episodic memory and proves its effectiveness through simulation results.

Increased corticospinal inhibition following brief maximal and submaximal contractions in humans.

A potentiating conditioning contraction (CC) has been shown to increase silent period duration, an index of corticospinal inhibition; however, it is unknown if the CC must induce potentiation for corticospinal inhibition to increase. Ten healthy, young adults (four females) completed this study to assess potentiation and silent period (SP) duration before and after four types of CCs: voluntary and electrically evoked maximal CCs to optimize potentiation, and voluntary and electrically evoked submaximal CCs (∼40% of maximal voluntary force) that induced minimal potentiation. Stimulation was applied to the ulnar nerve to evoke twitches for the assessment of potentiation and to evoke tetanic CCs of the first dorsal interosseous muscle. The SP was elicited by applying transcranial magnetic stimulation to the motor cortex during brief contractions at 25% of maximal voluntary force. Changes to twitch force and SP duration were not different for voluntary and tetanic contractions, so data were pooled. Twitch force increased by 81.2 ± 35.7% (P < 0.001) and 3.2 ± 6.5% (P = 0.039) following maximal and submaximal CCs, respectively. The SP was prolonged following maximal (12.6 ± 6.3%; P < 0.001) and submaximal (4.8 ± 4.9%; P < 0.001) CCs. Correlations between post-CC twitch force and SP duration were not significant for maximal or submaximal conditions (r = -0.068; r = 0.067; P ≥ 0.780, respectively). Duration of the SP increased not only following maximal-intensity CCs but also after submaximal-intensity CCs that induced virtually no potentiation (∼3%). Thus, we suggest that corticospinal inhibition is not directly related to mechanisms of muscle potentiation per se, but, rather, the level of muscle contraction likely mediates feedback from large diameter afferents that affect the SP.NEW & NOTEWORTHY The transcranial magnetic stimulation-induced silent period reflects a transient state of corticospinal inhibition that is influenced by recent history of muscle activation, which may include an effect of potentiation. We demonstrate that silent period duration increases following both voluntary and electrically evoked maximal and submaximal conditioning contractions, even though the latter intensity produced virtually no muscle potentiation. Feedback from group Ia and Ib muscle afferents is proposed as the cause of the increased corticospinal inhibition.

Postactivation Potentiation and the Asynchronous Action of Muscular and Neural Responses.

This study examined the underlying mechanisms of postactivation potentiation and the time course of muscular- and neural-related variables. Fourteen trained males executed 4 sets of six 6-second maximum isometric conditioning plantar flexions, with 15seconds and 2minutes of interval between the contractions and sets, respectively. Peak twitch torque (TT), rate of torque development, time to peak torque, half relaxation time, and the neural-related variables of H-reflex and electromyogram, normalized to the maximum M-wave (H/M and RMS/M, respectively), were evaluated, as well as the level of the voluntary activation, assessed by the twitch interpolation technique. All neural-related variables were analyzed for the trial within each set when TT was maximal and for the trial within each set when the neural-related variable itself was maximal. Compared with the baseline measures, TT and rate of torque development significantly increased in all sets (P < .001), whereas time to peak torque and half relaxation time significantly decreased in sets 1 to 4 and 2 to 4, respectively (P < .001). However, H/M and the RMS/M did not change for the repetition of each set for which the TT was maximal (P > .05). Interestingly, the within-set maximum H/M ratio of the lateral gastrocnemius muscle revealed a significant increase in all sets (P < .05), compared with the baseline measures. One set of 4 contractions with 6-second duration is sufficient to cause postactivation potentiation for most participants, whereas peak TT augmentation does not coincide with changes in the examined neural-related variables. Further experiments should consider the time lag on their maximal values and their inherent between-participants variability.

Mechanistic insights into cAMP-mediated presynaptic potentiation at hippocampal mossy fiber synapses.

Presynaptic plasticity is an activity-dependent change in the neurotransmitter release and plays a key role in dynamic modulation of synaptic strength. Particularly, presynaptic potentiation mediated by cyclic adenosine monophosphate (cAMP) is widely seen across the animals and thought to contribute to learning and memory. Hippocampal mossy fiber-CA3 pyramidal cell synapses have been used as a model because of robust presynaptic potentiation in short- and long-term forms. Moreover, direct presynaptic recordings from large mossy fiber terminals allow one to dissect the potentiation mechanisms. Recently, super-resolution microscopy and flash-and-freeze electron microscopy have revealed the localizations of release site molecules and synaptic vesicles during the potentiation at a nanoscale, identifying the molecular mechanisms of the potentiation. Incorporating these growing knowledges, we try to present plausible mechanisms underlying the cAMP-mediated presynaptic potentiation.

A step toward restoring hand functions in patients with multiple sclerosis-a study protocol.

Multiple sclerosis (MS) is a chronic autoimmune disease characterized by inflammation, demyelination of axons, and oligodendrocyte loss in the central nervous system. This leads to neurological dysfunction, including hand impairment, which is prevalent among patients with MS. However, hand impairment is the least targeted area for neurorehabilitation studies. Therefore, this study proposes a novel approach to improve hand functions compared to current strategies. Studies have shown that learning new skills in the motor cortex (M1) can trigger the production of oligodendrocytes and myelin, which is a critical mechanism for neuroplasticity. Transcranial direct current stimulation (tDCS) has been used to enhance motor learning and function in human subjects. However, tDCS induces non-specific effects, and concurrent behavioral training has been found to optimize its benefits. Recent research indicates that applying tDCS during motor learning can have priming effects on the long-term potentiation mechanism and prolong the effects of motor training in health and disease. Therefore, this study aims to assess whether applying repeated tDCS during the learning of a new motor skill in M1 can be more effective in improving hand functions in patients with MS than current neurorehabilitation strategies. If this approach proves successful in improving hand functions in patients with MS, it could be adopted as a new approach to restore hand functions. Additionally, if the application of tDCS demonstrates an accumulative effect in improving hand functions in patients with MS, it could provide an adjunct intervention during rehabilitation for these patients. This study will contribute to the growing body of literature on the use of tDCS in neurorehabilitation and could have a significant impact on the quality of life of patients with MS.

Efficient degradation of benzalkonium chloride by FeMn-CA300 catalyst activated persulfate process: Surface hydroxyl potentiation mechanism and degradation pathway

The consumption of disinfectants increased dramatically with the outbreak of the COVID-19 epidemic. Benzalkonium chloride (DDBAC), a cationic surfactant disinfectant for import and export cargoes, is used for effective degradation method. For DDBAC effective degradation, polyhedral Fe–Mn bimetallic catalyst of Prussian blue analogue (FeMn-CA300) was novelty developed for rapid peroxymonosulfate (PMS) activation. Results showed that the Fe/Mn redox and surface hydroxyl groups in the catalyst played an important role in the DDBAC-enhanced degradation. The removal effectiveness of 10 mg L−1 DDBAC was up to 99.4% in 80 min under the initial pH = 7, catalyst dosage of 0.4 g L−1, and PMS concentration of 15 mmol L−1. In addition, FeMn-CA300 had a wide pH applicability range. The results indicated that hydroxyls, sulfate radicals, and singlet oxygen could effectively improve the degradation efficiency, where sulfate radicals played a crucial role. Finally, the corresponding degradation path of DDBAC was further provided according to GC-MS results. The results of this study provide new insights into the degradation of DDBAC, thereby highlighting the great potential of FeMnca300/PMS to control refractory organic compounds in the aqueous phase.

Long-term potentiation mechanism of biological postsynaptic activity in neuro-inspired halide perovskite memristors

Perovskite memristors have emerged as leading contenders in brain-inspired neuromorphic electronics. Although these devices have been shown to accurately reproduce synaptic dynamics, they pose challenges for in-depth understanding of the underlying nonlinear phenomena. Potentiation effects on the electrical conductance of memristive devices have attracted increasing attention from the emerging neuromorphic community, demanding adequate interpretation. Here, we propose a detailed interpretation of the temporal dynamics of potentiation based on nonlinear electrical circuits that can be validated by impedance spectroscopy. The fundamental observation is that the current in a capacitor decreases with time; conversely, for an inductor, it increases with time. There is no electromagnetic effect in a halide perovskite memristor, but ionic-electronic coupling creates a chemical inductor effect that lies behind the potentiation property. Therefore, we show that beyond negative transients, the accumulation of mobile ions and the eventual penetration into the charge-transport layers constitute a bioelectrical memory feature that is the key to long-term synaptic enhancement. A quantitative dynamical electrical model formed by nonlinear differential equations explains the memory-based ionic effects to inductive phenomena associated with the slow and delayed currents, invisible during the ‘off mode’ of the presynaptic spike-based stimuli. Our work opens a new pathway for the rational development of material mimesis of neural communications across synapses, particularly the learning and memory functions in the human brain, through a Hodgkin–Huxley-style biophysical model.

Passivation Improvement of Poly‐SiOx Based TOPCon Contacts by Thermal Annealing in a Water Vapor Atmosphere: Mechanism Exploration and Application Research

AbstractPost‐treatment techniques of tunnel oxide passivated contact (TOPCon) structure are universally implemented via executing an additional hydrogenation process to optimize the passivation performance. However, the underlying physical mechanism and which method is most applicable are still being investigated. Herein, the effectiveness of thermal annealing in water vapor and N2 atmosphere is studied, which is both environmentally friendly and easy to operate. It is demonstrated that compared to other common hydrogenation techniques, the wet N2 outperforms in improving the passivation performance, which can be attributed to the neutralization of internal defects in poly‐Si and the optimization of structural densities, and interestingly, this gain effect is amplified when this contact is doped with oxygen impurity. A power conversion efficiency of 22.62% is achieved using this technology which verifies its reliability and applicability. A loss analysis based on numerical simulations, outlining ways to achieve higher conversion efficiency and highlighting the great potential of this technology is also provided. Extensive experiments and first‐principles calculations based on density‐functional theory are conducted to clarify the underlying dynamics, including the surface adsorption process and the potentiation mechanisms, revealing that passivation and neutralization of hydrogen atoms couple with the compactness optimization of the structure.

Short-term plasticity of the motor cortex compensates for bradykinesia in Parkinson’s disease

Patients with Parkinson's disease (PD) show impaired short-term potentiation (STP) mechanisms in the primary motor cortex (M1). However, the role played by this neurophysiological abnormality in bradykinesia pathophysiology is unknown. In this study, we used a multimodal neuromodulation approach to test whether defective STP contributes to bradykinesia. We evaluated STP by measuring motor-evoked potential facilitation during 5 Hz-repetitive transcranial magnetic stimulation (rTMS) and assessed repetitive finger tapping movements through kinematic techniques. Also, we used transcranial alternating current stimulation (tACS) to drive M1 oscillations and experimentally modulate bradykinesia. STP was assessed during tACS delivered at beta (β) and gamma (γ) frequency, and during sham-tACS. Data were compared to those recorded in a group of healthy subjects. In PD, we found that STP was impaired during sham- and γ-tACS, while it was restored during β-tACS. Importantly, the degree of STP impairment was associated with the severity of movement slowness and amplitude reduction. Moreover, β-tACS-related improvements in STP were linked to changes in movement slowness and intracortical GABA-A-ergic inhibition during stimulation, as assessed by short-interval intracortical inhibition (SICI). Patients with prominent STP amelioration had greater SICI reduction (cortical disinhibition) and less slowness worsening during β-tACS. Dopaminergic medications did not modify β-tACS effects. These data demonstrate that abnormal STP processes are involved in bradykinesia pathophysiology and return to normal levels when β oscillations increase. STP changes are likely mediated by modifications in GABA-A-ergic intracortical circuits and may represent a compensatory mechanism against β-induced bradykinesia in PD.

Synthetic Multivalent Disulfide-Constrained Peptide Agonists Potentiate Wnt1/β-Catenin Signaling via LRP6 Coreceptor Clustering.

Wnt ligands are critical for tissue homeostasis and form a complex with LRP6 and frizzled coreceptors to initiate Wnt/β-catenin signaling. Yet, how different Wnts achieve various levels of signaling activation through distinct domains on LRP6 remains elusive. Developing tool ligands that target individual LRP6 domains could help elucidate the mechanism of Wnt signaling regulation and uncover pharmacological approaches for pathway modulation. We employed directed evolution of a disulfide constrained peptide (DCP) to identify molecules that bind to the third β-propeller domain of LRP6. The DCPs antagonize Wnt3a while sparing Wnt1 signaling. Using PEG linkers with different geometries, we converted the Wnt3a antagonist DCPs to multivalent molecules that potentiated Wnt1 signaling by clustering the LRP6 coreceptor. The mechanism of potentiation is unique as it occurred only in the presence of extracellular secreted Wnt1 ligand. While all DCPs recognized a similar binding interface on LRP6, they displayed different spatial orientations that influenced their cellular activities. Moreover, structural analyses revealed that the DCPs exhibited new folds that were distinct from the parent DCP framework they were evolved from. The multivalent ligand design principles highlighted in this study provide a path for developing peptide agonists that modulate different branches of cellular Wnt signaling.

Effects of Hydrostatic-Pressure on Muscle Contraction: A Look Back on Some Experimental Findings.

Findings from experiments that used hydrostatic pressure changes to analyse the process of skeletal muscle contraction are re-examined. The force in resting muscle is insensitive to an increase in hydrostatic pressure from 0.1 MPa (atmospheric) to 10 MPa, as also found for force in rubber-like elastic filaments. The force in rigour muscle rises with increased pressure, as shown experimentally for normal elastic fibres (e.g., glass, collagen, keratin, etc.). In submaximal active contractions, high pressure leads to tension potentiation. The force in maximally activated muscle decreases with increased pressure: the extent of this force decrease in maximal active muscle is sensitive to the concentration of products of ATP hydrolysis (Pi-inorganic phosphate and ADP-adenosine diphosphate) in the medium. When the increased hydrostatic pressure is rapidly decreased, the force recovered to the atmospheric level in all cases. Thus, the resting muscle force remained the same: the force in the rigour muscle decreased in one phase and that in active muscle increased in two phases. The rate of rise of active force on rapid pressure release increased with the concentration of Pi in the medium, indicating that it is coupled to the Pi release step in the ATPase-driven crossbridge cycle in muscle. Pressure experiments on intact muscle illustrate possible underlying mechanisms of tension potentiation and causes of muscle fatigue.

Mechanistic insights into lipid regulation of the acid-sensing ion channels.

Acid-sensing ion channels are primary sensors of acidosis in the central and peripheral nervous systems. In this respect, ASICs have been implicated in a number of pathologic conditions where tissue acidosis occurs, including pain sensing during inflammation. Interestingly, although ASICs are activated by protons, the prolonged acidosis present in these conditions should render the channels inactive. One potential solution to this paradox is that, in addition to elevated concentration of protons, there are elevated endogenous inflammatory molecules that further potentiate the channel. Among the molecules elevated in inflammatory conditions are single acyl chain lipids like the polyunsaturated fatty acids (PUFAs). We have previously demonstrated a number of these lipids can stabilize the open state of ASICs, leading to a prolonged non-desensitizing activation, which in some cases can occur even at neutral pH. We found that this effect was dependent on a negatively charged head interaction with the outer transmembrane domain of the channel. However, a detailed understanding of the residues involved in this interaction and the mechanism by which lipids potentiate ASICs remain unknown. Here, we explore these open questions using patch-clamp electrophysiology. We find that lipid potentiation of the channel occurs in a state-dependent manner. We highlight isoform-dependent effects of lipid action on ASICs and use site-directed mutagenesis to narrow down key residues important in lipid binding. We examine lipid effects on the single channel parameters to determine how potentiation manifests at the whole-cell level. Finally, we test several potential mechanisms by which lipids potentiate, including the potential alteration of ionic concentrations at activation sites. Together, we build a model of the binding and mechanism of potentiation for an endogenous regulator of ASICs and provide a basis for future therapeutic development.

The CNS-penetrating taxane drug TPI 287 potentiates antiglioma activity of the AURKA inhibitor alisertib in vivo.

Glioblastoma (GBM) has a very poor prognosis despite current treatment. We previously found cytotoxic synergy between the AURKA inhibitor alisertib and the CNS-penetrating taxane TPI 287 against GBM tumor cells in vitro. We used an orthotopic human GBM xenograft mouse model to test if TPI 287 potentiates alisertib in vivo. Western blotting, immunohistochemistry, siRNA knockdown, annexin V binding, and 3-dimensional Matrigel invasion assays were used to investigate potential mechanisms of alisertib and TPI 287 treatment interactions. Alisertib + TPI 287 combination therapy significantly prolonged animal survival compared to vehicle (p = 0.011), but only marginally compared to alisertib alone. Alisertib, TPI 287, and combined alisertib + TPI 287 reduced animal tumor volume compared to vehicle-treated controls. This was statistically significant for the combination therapy at 4 weeks (p < 0.0001). Alisertib + TPI 287 treatment decreased anti-apoptotic Bcl-2 protein levels in vivo and in vitro. Expression of the pro-apoptotic protein Bak was significantly increased by combination treatment (p < 0.0001). Pro-apoptotic Bim and Bak knockdown by siRNA decreased apoptosis by alisertib + TPI 287 in GB9, GB30, and U87 cells (p = 0.0005 to 0.0381). Although alisertib and TPI 287 significantly reduced GBM cell invasion (p < 0.0001), their combination was no more effective than TPI 287 alone. Results suggest that apoptosis is the dominant mechanism of potentiation of GBM growth inhibition by alisertib + TPI 287, in part through effects on Bcl-2 family proteins, providing a rationale for further laboratory testing of an AURKA inhibitor plus TPI 287 as a potential therapy against GBM.

Mechanisms That Underlie Expression of Estradiol-Induced Excitatory Synaptic Potentiation in the Hippocampus Differ between Males and Females.

17β-estradiol (E2) is synthesized in the hippocampus of both sexes and acutely potentiates excitatory synapses in each sex. Previously, we found that the mechanisms for initiation of E2-induced synaptic potentiation differ between males and females, including in the molecular signaling involved. Here, we used electrical stimulation and two-photon glutamate uncaging in hippocampal slices from adult male and female rats to investigate whether the downstream consequences of distinct molecular signaling remain different between the sexes or converge to the same mechanism(s) of expression of potentiation. This showed that synaptic activity is necessary for expression of E2-induced potentiation in females but not males, which paralleled a sex-specific requirement in females for calcium-permeable AMPARs (cpAMPARs) to stabilize potentiation. Nonstationary fluctuation analysis of two-photon evoked unitary synaptic currents showed that the postsynaptic component of E2-induced potentiation occurs either through an increase in AMPAR conductance or in nonconductive properties of AMPARs (number of channels × open probability) and never both at the same synapse. In females, most synapses (76%) were potentiated via increased AMPAR conductance, whereas in males, more synapses (60%) were potentiated via an increase in nonconductive AMPAR properties. Inhibition of cpAMPARs eliminated E2-induced synaptic potentiation in females, whereas some synapses in males were unaffected by cpAMPAR inhibition; these synapses in males potentiated exclusively via increased AMPAR nonconductive properties. This sex bias in expression mechanisms of E2-induced synaptic potentiation underscores the concept of latent sex differences in mechanisms of synaptic plasticity in which the same outcome in each sex is achieved through distinct underlying mechanisms.SIGNIFICANCE STATEMENT Estrogens are synthesized in the brains of both sexes and potentiate excitatory synapses to the same degree in each sex. Despite this apparent similarity, the molecular signaling that initiates estrogen-induced synaptic potentiation differs between the sexes. Here we show that these differences extend to the mechanisms of expression of synaptic potentiation and result in distinct patterns of postsynaptic neurotransmitter receptor modulation in each sex. Such latent sex differences, in which the same outcome is achieved through distinct underlying mechanisms in males versus females, indicate that molecular mechanisms targeted for drug development may differ between the sexes even in the absence of an overt sex difference in behavior or disease.