bluuebunny/crossref_metadata_2025 huggingface dataset:
Plastic Changes in Nociceptive Transmission of the Rat Spinal Cord With Advancing Age
Authors: Iwata Koichi , Fukuoka Tetsuo , Kondo Eji , Tsuboi Yoshiyuki , Tashiro Akimasa , Noguchi Koichi , Masuda Yuji , Morimoto Toshifumi , Kanda Kenro
Published: May 2017

To understand characteristics of the pain system in the elderly, we investigated the electrophysiological properties of nociceptive neurons in the lumbar spinal dorsal horn of aged (29–34-mo old) and adult (7–13-mo old) rats. The responses of nociceptive neurons to noxious thermal stimulation, as well as the spontaneous firing rate, were significantly higher in the aged as compared with adult rats. Furthermore, the size of the high-threshold receptive field area of wide dynamic range neurons was larger ( P < 0.01) and that of the low-threshold area was smaller ( P< 0.05) in aged rats than in adult rats. The increased nociceptive neuronal activity in the aged rats correlated with the finding that the paw withdrawal latency was significantly shorter in the aged rats than those of the adult rats following heat stimulation of the hind paw ( P < 0.05). Reversible local anesthetic block of descending pathways resulted in a dramatic increase in neuronal activity in adult rats but had little effect in aged rats. There was also a significant loss of serotoninergic and noradrenergic fibers in the spinal dorsal horn of the aged rats. These results demonstrate an age-related plasticity in spinal nociceptive processing that is related to impairment of descending modulatory pathways.
Processing of Nociceptive Mechanical and Thermal Information in Central Amygdala Neurons With Knee-Joint Input
Authors: Neugebauer Volker , Li Weidong
Published: May 2017

Pain has a strong emotional dimension, and the amygdala plays a key role in emotionality. The processing of nociceptive mechanical and thermal information was studied in individual neurons of the central nucleus of the amygdala, the target of the spino-parabrachio-amygdaloid pain pathway and a major output nucleus of the amygdala. This study is the first to characterize nociceptive amygdala neurons with input from deep tissue, particularly the knee joint. In 46 anesthetized rats, extracellular single-unit recordings were made from 119 central amygdala neurons that were activated orthodromically by electrical stimulation in the lateral pontine parabrachial area and were tested for receptive fields in the knee joints. Responses to brief mechanical stimulation of joints, muscles, and skin and to cutaneous thermal stimuli were recorded. Receptive-field sizes and thresholds were mapped and stimulus-response functions constructed. Neurons in the central nucleus of the amygdala with excitatory input from the knee joint ( n = 62) typically had large symmetrical receptive fields in both hindlimbs or in all four extremities and responded exclusively or preferentially to noxious mechanical stimulation of deep tissue ( n = 58). Noxious mechanical stimulation of the skin excited 30 of these neurons; noxious heat activated 21 neurons. Stimulus-response data were best fitted by a sigmoid nonlinear regression model rather than by a monotonically increasing linear function. Another 15 neurons were inhibited by noxious mechanical stimulation of the knee joint and other deep tissue. Fifteen neurons had no receptive field in the knee but responded to noxious stimulation of other body areas; 27 nonresponsive neurons were not activated by natural somesthetic stimulation. Our data suggest that excitation is the predominant effect of brief painful stimulation of somatic tissue on the population of central amygdala neurons with knee joint input. Their large symmetrical receptive fields and sigmoid rather than monotonically increasing linear stimulus-response functions suggest a role of nociceptive central amygdala neurons in other than sensory-discriminative aspects of pain.
mAChRs in the Grasshopper Brain Mediate Excitation by Activation of the AC/PKA and the PLC Second-Messenger Pathways
Authors: Wenzel B. , Elsner N. , Heinrich R.
Published: May 2017

The species-specific sound production of acoustically communicating grasshoppers can be stimulated by pressure injection of both nicotinic and muscarinic agonists into the central body complex and a small neuropil situated posterior and dorsal to it. To determine the role of muscarinic acetylcholine receptors (mAChRs) in the control of acoustic communication behavior and to identify the second-messenger pathways affected by mAChR-activation, muscarinic agonists and membrane-permeable drugs known to interfere with specific mechanisms of intracellular signaling pathways were pressure injected to identical sites in male grasshopper brains. Repeated injections of small volumes of muscarine elicited stridulation of increasing duration associated with decreased latencies. This suggested an accumulation of excitation over time that is consistent with the suggested role of mAChRs in controling courtship behavior: to provide increasing arousal leading to higher intensity of stridulation and finally initiating a mating attempt. At sites in the brain where muscarine stimulation was effective, stridulation could be evoked by forskolin, an activator of adenylate cyclase (AC); 8-Br-cAMP-activating protein kinase A (PKA); and 3-isobuty-1-methylxanthine, leading to the accumulation of endogenously generated cAMP through inhibition of phosphodiesterases. This suggested that mAChRs mediate excitation by stimulating the AC/cAMP/PKA pathway. In addition, muscarine-stimulated stridulation was inhibited by 2′-5′-dideoxyadenonsine and SQ 22536, two inhibitors of AC; H-89 and Rp-cAMPS, two inhibitors of PKA; and by U-73122 and neomycin, two agents that inhibit phospholipase C (PLC) by independent mechanisms. Because the inhibition of AC, PKA, or PLC by various individually applied substances entirely suppressed muscarine-evoked stridulation in a number of experiments, activation of both pathways, AC/cAMP/PKA and PLC/IP3/diacylglycerine, appeared to be necessary to mediate the excitatory effects of mAChRs. With these studies on an intact “behaving” grasshopper preparation, we present physiological relevance for mAChR-evoked excitation mediated by sequential activation of the AC- and PLC-initiated signaling pathways that has been reported in earlier in vitro studies.
Modifications of Vestibular Responses of Individual Reticulospinal Neurons in Lamprey Caused by Unilateral Labyrinthectomy
Authors: Deliagina T. G. , Pavlova E. L.
Published: May 2017

A postural control system in the lamprey is driven by vestibular input and maintains the dorsal-side-up orientation of the animal during swimming. After a unilateral labyrinthectomy (UL), the lamprey continuously rolls toward the damaged side. Normally, a recovery of postural equilibrium (“vestibular compensation”) takes about 1 mo. However, illumination of the eye contralateral to UL results in an immediate and reversible restoration of equilibrium. Here we used eye illumination as a tool to examine a functional recovery of the postural network. Important elements of this network are the reticulospinal (RS) neurons, which are driven by vestibular input and transmit commands for postural corrections to the spinal cord. In this study, we characterized modifications of the vestibular responses in individual RS neurons caused by UL and the effect exerted on these responses by eye illumination. The activity of RS neurons was recorded from their axons in the spinal cord by chronically implanted electrodes, and spikes in individual axons were extracted from the population activity signals. The same neurons were recorded both before and after UL. Vestibular stimulation (rotation in the roll plane through 360°) and eye illumination were performed in quiescent animals. It was found that the vestibular responses on the UL-side changed only slightly, whereas the responses on the opposite side disappeared almost completely. This asymmetry in the bilateral activity of RS neurons is the most likely cause for the loss of equilibrium in UL animals. Illumination of the eye contralateral to UL resulted, first, in a restoration of vestibular responses in the neurons inactivated by UL and in an appearance of vestibular responses in some other neurons that did not respond to vestibular input before UL. These responses had directional sensitivity and zones of spatial sensitivity similar to those observed before UL. However, their magnitude was smaller than before UL. Second, the eye illumination caused a reduction of the magnitude of vestibular responses on the UL side. These two factors tend to restore symmetry in bilateral activity of RS neurons, which is the most likely cause for the recovery of equilibrium in the swimming UL lamprey. Results of this study are discussed in relation to the model of the roll control system proposed in our previous studies as well as in relation to the vestibular compensation.
NMDA-Induced Calcium Loads Recycle Across the Mitochondrial Inner Membrane of Hippocampal Neurons in Culture
Authors: Wang Guang Jian , Thayer Stanley A.
Published: May 2017

Mitochondria sequester N-methyl-d-aspartate (NMDA)-induced Ca2+ loads and regulate the shape of intracellular Ca2+ concentration ([Ca2+]i) responses in neurons. When isolated mitochondria are exposed to high [Ca2+],Ca2+ enters the matrix via the uniporter and returns to the cytosol by Na+/Ca2+ exchange. Released Ca2+ may re-enter the mitochondrion recycling across the inner membrane dissipating respiratory energy. Ca2+ recycling, the continuous uptake and release of Ca2+ by mitochondria, has not been described in intact neurons. Here we used single-cell microfluorimetry to measure [Ca2+]i and mitochondrially targeted aequorin to measure matrix Ca2+ concentration ([Ca2+]mt) to determine whether Ca2+ recycles across the mitochondrial inner membrane in intact neurons following treatment with NMDA. We used ruthenium red and CGP 37157 to block uptake via the uniporter and release via Na+/Ca2+exchange, respectively. As predicted by the Ca2+recycling hypothesis, blocking the uniporter immediately following challenge with 200 μM NMDA produced a rapid and transient increase in cytosolic Ca2+ without a corresponding increase in matrix Ca2+. Blocking mitochondrial Ca2+ release produced the opposite effect, depressing cytosolic Ca2+ levels and prolonging the time for matrix Ca2+ levels to recover. The Ca2+ recycling hypothesis uniquely predicts these reciprocal changes in the Ca2+ levels between the two compartments. Ca2+ recycling was not detected following treatment with 20 μM NMDA. Thus Ca2+recycling across the inner membrane was more pronounced following treatment with a high relative to a low concentration of NMDA, consistent with a role in Ca2+-dependent neurotoxicity.
Neural Correlates of Tasting Concentrated Quinine and Sugar Solutions
Authors: Zald David H. , Hagen Mathew C. , Pardo José V.
Published: May 2017

Behavioral, ethological, and electrophysiological evidence suggests that the highly unpleasant, bitter taste of a concentrated quinine hydrochloride (QHCL) should activate the human amygdala. In the present study, healthy subjects tasted 0.02 M QHCL or water while regional cerebral blood flow (rCBF) was assayed with H2 15O PET. Subjects were also studied while tasting a pleasant sucrose solution and resting with eyes closed (ECR). Tasting QHCL significantly increased rCBF within the left amygdala relative to control conditions of tasting water and ECR. Sucrose and water caused small to moderate rCBF increases in the amygdala relative to ECR, but sucrose did not significantly increase activity within either amygdalae relative to water. In the frontal lobe, QHCL and sucrose both activated the right posterior orbitofrontal cortex (OFC) relative to water, but portions of the anterior OFC and inferior frontal pole showed valence specific responses to QHCL. These data indicate that the left amygdala responds robustly to QHCL and more moderately to nonaversive sapid stimuli, both pleasant and unpleasant gustatory stimuli activate the right posterior OFC, and the left inferior frontal pole/anterior OFC demonstrates valence-specific responses to aversive gustatory stimuli.
Novel Excitatory Actions of Galanin on Rat Cholinergic Basal Forebrain Neurons: Implications for Its Role in Alzheimer's Disease
Authors: Jhamandas Jack H. , Harris Kim H. , MacTavish David , Jassar Balvinder S.
Published: May 2017

Galanin, a 29-amino-acid neuropeptide, is generally viewed as an inhibitory neuromodulator in a variety of central systems. Galanin expression is upregulated in the cholinergic basal forebrain nuclei in Alzheimer's disease (AD) and is postulated to play an important role in memory and cognitive function. In this study, application of galanin to acutely dissociated rat neurons from the basal forebrain nucleus diagonal band of Broca (DBB), caused a decrease in whole cell voltage-activated currents in a majority of cells. Galanin reduces a suite of potassium currents, including calcium-activated potassium ( I C), the delayed rectifier ( I K), and transient outward potassium ( I A) conductances, but not calcium or sodium currents. Under current-clamp conditions, application of galanin evoked an increase in excitability and a loss of accommodation in cholinergic DBB neurons. Using single-cell RT-PCR technique, we determined that galanin actions were specific to cholinergic, but not GABAergic DBB neurons The notion that galanin plays a deleterious role in AD is based, in part, on galanin hyperinnervation of cholinergic cells in the basal forebrain of AD patients, its ability to depress acetylcholine release and its inhibitory actions at other CNS sites. However, our results suggest that by virtue of its excitatory actions on cholinergic neurons, galanin may in fact play a compensatory role by augmenting the release of acetylcholine from remaining cholinergic basal forebrain neurons. This action might serve to delay the progression of AD pathology linked to a reduction in central cholinergic tone.
Neural Noise Can Explain Expansive, Power-Law Nonlinearities in Neural Response Functions
Authors: Miller Kenneth D. , Troyer Todd W.
Published: May 2017

Many phenomenological models of the responses of simple cells in primary visual cortex have concluded that a cell's firing rate should be given by its input raised to a power greater than one. This is known as an expansive power-law nonlinearity. However, intracellular recordings have shown that a different nonlinearity, a linear-threshold function, appears to give a good prediction of firing rate from a cell's low-pass-filtered voltage response. Using a model based on a linear-threshold function, Anderson et al. showed that voltage noise was critical to converting voltage responses with contrast-invariant orientation tuning into spiking responses with contrast-invariant tuning. We present two separate results clarifying the connection between noise-smoothed linear-threshold functions and power-law nonlinearities. First, we prove analytically that a power-law nonlinearity is the only input-output function that converts contrast-invariant input tuning into contrast-invariant spike tuning. Second, we examine simulations of a simple model that assumes instantaneous spike rate is given by a linear-threshold function of voltage and voltage responses include significant noise. We show that the resulting average spike rate is well described by an expansive power law of the average voltage (averaged over multiple trials), provided that average voltage remains less than about 1.5 SDs of the noise above threshold. Finally, we use this model to show that the noise levels recorded by Anderson et al. are consistent with the degree to which the orientation tuning of spiking responses is more sharply tuned relative to the orientation tuning of voltage responses. Thus neuronal noise can robustly generate power-law input-output functions of the form frequently postulated for simple cells.
Properties of Primary Sensory (Lemniscal) Synapses in the Ventrobasal Thalamus and the Relay of High-Frequency Sensory Inputs
Authors: Castro-Alamancos Manuel A.
Published: May 2017

The main role of the thalamus is to relay sensory inputs to the neocortex. In the primary somatosensory thalamus (ventrobasal thalamus), sensory inputs deliver tactile information through the medial lemniscus tract. The transmission of sensory information through this pathway is affected by behavioral state. For instance, the relay of high-frequency somatosensory inputs through the thalamus is suppressed during anesthesia or quiescent states but allowed during behaviorally activated states. This change may be due to the effects of modulators on the efficacy of lemniscal synapses. Here I show that lemniscal synapses of adult rodents studied in vitro produce large amplitude-highly secure unitary excitatory postsynaptic potentials (EPSPs), which depress in response to repetitive stimulation at frequencies >2 Hz. Acetylcholine and norepinephrine, which are important thalamic modulators, have no effect on the efficacy of lemniscal EPSPs but reduce evoked inhibitory postsynaptic potentials and corticothalamic EPSPs. Although acetylcholine and norepinephrine do not affect lemniscal synapses, the postsynaptic depolarization they produce on thalamocortical neurons serves to warrant the relay of lemniscal inputs at high-frequency rates by bringing the depressed lemniscal EPSPs close to firing threshold. In conclusion, acetylcholine and norepinephrine released during activated states selectively enhance sensory transmission through the lemniscal pathway by depolarizing thalamocortical neurons and simultaneously depressing the other afferent pathways.
Differential Effect of Ethanol on NMDA EPSCs in Pyramidal Cells in the Posterior Cingulate Cortex of Juvenile and Adult Rats
Authors: Li Qiang , Wilson Wilkie A. , Swartzwelder H. Scott
Published: May 2017

Ethanol (EtOH) is a potent inhibitor of N-methyl-d-aspartate (NMDA) receptor–mediated activity in a number of brain areas, and recent studies have indicated that this inhibitory effect of ethanol is more powerful in the juvenile brain compared with the adult brain. However, previous direct developmental comparisons have been limited to studies of extracellular responses in the hippocampus. To begin an assessment of the mechanisms underlying this developmental sensitivity, we assessed the inhibitory effect of EtOH on NMDA receptor–mediated synaptic transmission in neocortical slices from adult (95–135 days old) and juvenile (28–32 days old) rats using the whole cell patch-clamp recording technique. In the presence of 6,7-dinitroquinoxaline-2,3-dione (20 μM) and bicuculline methiodine (20 μM), NMDA receptor–mediated excitatory postsynaptic currents were isolated from pyramidal cells of the posterior cingulate cortex (PCC). In slices from juvenile rats 5, 10, 30, and 60 mM EtOH reduced the mean amplitude of NMDA receptor–mediated EPSCs by 11, 22, 35, and 46%, respectively. However, the same concentrations of EtOH inhibited the mean amplitude of EPSCs by only 4, 8, 15, and 31% in slices from adult rats. This developmental difference in the potency of EtOH against NMDA receptor–mediated EPSCs was also observed when the holding potential of the neurons was increased to +30 mV, although the inhibitory effect of ethanol on adult neurons was diminished at that voltage. These results provide a cellular analysis of the enhanced potency of ethanol against NMDA receptor–mediated EPSCs in neocortical cells from juvenile animals compared with adults.
Gaze-, Eye-, and Head-Movement Dynamics During Closed- and Open-Loop Gaze Pursuit
Authors: Dubrovsky Alexander S. , Cullen Kathleen E.
Published: May 2017

Horizontal step-ramp stimuli were used to examine gaze-, eye-, and head-movement dynamics during head-unrestrained pursuit in two rhesus monkeys. In a first series of experiments, we characterized and compared head-restrained (HR) and -unrestrained (HU) pursuit responses to unpredictable, nonperiodic, constant velocity (20–80°/s) stimuli. When the head was free to move, both monkeys used a combination of eye and head motion to initially fixate and then pursue the target. The pursuit responses (i.e., gaze responses) were highly stereotyped and nearly identical among the HR and HU conditions for a given step-ramp stimulus. In the HU condition, initial eye and initial head acceleration tended to increase as a function of target velocity but did not vary systematically with initial target eccentricity. In a second series of experiments, step-ramp stimuli (40°/s) were presented, and, ∼125 ms after pursuit onset, a constant retinal velocity error (RVE) was imposed for a duration of 300 ms. In each monkey, HR and HU gaze velocity was similarly affected by stabilizing the target with respect to the monkey's fovea (i.e., RVE = 0°/s) and by moving the target with constant retinal velocity errors (i.e., RVE = ±10°/s). In the HU condition, changes in both eye and head velocity trajectories contributed to the observed gaze velocity responses to imposed RVEs. We conclude that eye and head movements are not independently controlled during HU pursuit but rather are controlled, at least in part, by a shared upstream controller within the pursuit pathways.
D2 Receptors Inhibit the Secretory Process Downstream From Calcium Influx in Dopaminergic Neurons: Implication of K+ Channels
Authors: Congar Patrice , Bergevin Annie , Trudeau Louis-Eric
Published: May 2017

Dopaminergic (DAergic) neurons possess D2-like somatodendritic and terminal autoreceptors that modulate cellular excitability and dopamine (DA) release. The cellular and molecular processes underlying the rapid presynaptic inhibition of DA release by D2 receptors remain unclear. Using a culture system in which isolated DAergic neurons establish self-innervating synapses (“autapses”) that release both DA and glutamate, we studied the mechanism by which presynaptic D2 receptors inhibit glutamate-mediated excitatory postsynaptic currents (EPSCs). Action-potential evoked EPSCs were reversibly inhibited by quinpirole, a selective D2 receptor agonist. This inhibition was slightly reduced by the inward rectifier K+ channel blocker barium, largely prevented by the voltage-dependent K+channel blocker 4-aminopyridine, and completely blocked by their combined application. The lack of a residual inhibition of EPSCs under these conditions argues against the implication of a direct inhibition of presynaptic Ca2+ channels. To evaluate the possibility of a direct inhibition of the secretory process, spontaneous miniature EPSCs were evoked by the Ca2+ ionophore ionomycin. Ionomycin-evoked release was insensitive to cadmium and dramatically reduced by quinpirole, providing evidence for a direct inhibition of quantal release at a step downstream to Ca2+ influx through voltage-dependent Ca2+ channels. Surprisingly, this effect of quinpirole on ionomycin-evoked release was blocked by 4-aminopyridine. These results suggest that D2 receptor activation decreases neurotransmitter release from DAergic neurons through a presynaptic mechanism in which K+ channels directly inhibit the secretory process.
Physiological Properties of Late Inspiratory Neurons and Their Possible Involvement in Inspiratory Off-Switching in Cats
Authors: Haji Akira , Okazaki Mari , Yamazaki Hiromi , Takeda Ryuji
Published: May 2017

To assess the functional significance of late inspiratory (late-I) neurons in inspiratory off-switching (IOS), membrane potential and discharge properties were examined in vagotomized, decerebrate cats. During spontaneous IOS, late-I neurons displayed large membrane depolarization and associated discharge of action potentials that started in late inspiration, peaked at the end of inspiration, and ended during postinspiration. Depolarization was decreased by iontophoresis of dizocilpine and eliminated by tetrodotoxin. Stimulation of the vagus nerve or the nucleus parabrachialis medialis (NPBM) also evoked depolarization of late-I neurons and IOS. Waves of spontaneous chloride-dependent inhibitory postsynaptic potentials (IPSPs) preceded membrane depolarization during early inspiration and followed during postinspiration and stage 2 expiration of the respiratory cycle. Iontophoresed bicuculline depressed the IPSPs. Intravenous dizocilpine caused a greatly prolonged inspiratory discharge of the phrenic nerve (apneusis) and suppressed late-inspiratory depolarization as well as early-inspiratory IPSPs, resulting in a small constant depolarization throughout the apneusis. NPBM or vagal stimulation after dizocilpine produced small, stimulus-locked excitatory postsynaptic potentials (EPSPs) in late-I neurons. Neurobiotin-labeled late-I neurons revealed immunoreactivity for glutamic acid decarboxylase as well as N-methyl-d-aspartate (NMDA) receptors. These results suggest that late-I neurons are GABAergic inhibitory neurons, while the effects of bicuculline and dizocilpine indicate that they receive periodic waves of GABAergic IPSPs and glutamatergic EPSPs. The data lead to the conclusion that late-I neurons play an important inhibitory role in IOS. NMDA receptors are assumed to augment and/or synchronize late-inspiratory depolarization and discharge of late-I neurons, leading to GABA release and consequently off-switching of bulbar inspiratory neurons and phrenic motoneurons.
Temporal Modulation Transfer Functions in Cat Primary Auditory Cortex: Separating Stimulus Effects From Neural Mechanisms
Authors: Eggermont Jos J.
Published: May 2017

We present here a comparison between the local field potentials (LFP) and multiunit (MU) responses, comprising 401 single units, in primary auditory cortex (AI) of 31 cats to periodic click trains, gamma-tone and time-reversed gamma-tone trains, AM noise, AM tones, and frequency-modulated (FM) tones. In a large number of cases, the response to all six stimuli was obtained for the same neurons. We investigate whether cortical neurons are likely to respond to all types of repetitive transients and modulated stimuli and whether a dependence on modulating waveform, or tone or noise carrier, exists. In 97% of the recordings, a temporal modulation transfer function (tMTF) for MU activity was obtained for gamma-tone trains, in 92% for periodic click trains, in 83% for time-reversed gamma-tone trains, in 82% for AM noise, in 71% for FM tones, and only in 53% for AM tones. In 31% of the cases, the units responded to all six stimuli in an envelope-following way. These particular units had significantly larger onset responses to each stimulus compared with all other units. The overall response distribution shows the preference of AI units for stimuli with short rise times such as clicks and gamma tones. It also shows a clear asymmetry in the ability to respond to AM noise and AM tones and points to a strong effect of the frequency content of the carrier on the subcortical processing of AM stimuli. Yet all temporal response properties were independent of characteristic frequency and frequency-tuning curve bandwidth. We show that the observed differences in the tMTFs for different stimuli are to a large extent produced by the different degree of phase locking of the neuronal firings to the envelope of the first stimulus in the train or first modulation period. A normalization procedure, based on these synchronization differences, unified the tMTFs for all stimuli except clicks and allowed the identification of a largely stimulus-invariant, low-pass temporal filter function that most likely reflects the properties of synaptic depression and facilitation. For nonclick stimuli, the low-pass filter has a cutoff frequency of ∼10 Hz and a slope of ∼6 dB/octave. For nonclick stimuli, there was a systematic difference between the vector strength for LFPs and MU activity that can likely be attributed to postactivation suppression mechanisms.
Fast Network Oscillations in the Rat Dentate Gyrus In Vitro
Authors: Towers Stephen K. , LeBeau Fiona E. N. , Gloveli Tengis , Traub Roger D. , Whittington Miles A. , Buhl Eberhard H.
Published: May 2017

The dentate gyrus is a prominent source of gamma frequency activity in the hippocampal formation in vivo. Here we show that transient epochs of gamma frequency network activity (67 ± 12 Hz) can be generated in the dentate gyrus of rat hippocampal slices, following brief pressure ejections of a high-molarity potassium solution onto the molecular layer. Oscillatory activity remains synchronized over distances >300 μm and is accompanied by a modest rise in [K+]o. Gamma frequency oscillations were abolished by a GABAA receptor antagonist demonstrating their dependence on rhythmic inhibition. However, in many cases, higher frequency oscillations (>80 Hz) remained in the absence of synaptic transmission, thus demonstrating that nonsynaptic factors may underlie fast oscillatory activity.
Distributed and Concentration-Invariant Spatial Representations of Odorants by Receptor Neuron Input to the Turtle Olfactory Bulb
Authors: Wachowiak Matt , Cohen Lawrence B. , Zochowski Michal R.
Published: May 2017

We sought to characterize how odorants are represented at the level of afferent input to the olfactory bulb of the box turtle, a terrestrial reptile that, like mammals, detects airborne odorants. Using methods developed first in zebrafish, we selectively labeled olfactory receptor neurons with Calcium Green-1 dextran and imaged odorant-evoked input to glomeruli in vivo. Odorant representations were imaged at a glomerular level of resolution over a portion of the dorsal olfactory bulb and at a regional level of resolution over the entire dorsal surface. We report two new findings. First, even at low concentrations, odorants typically elicited input to a large fraction of all imaged glomeruli. Second, while the amplitude of the odorant-evoked input to glomeruli was concentration dependent, the relative pattern of input to the bulb changed only slightly over a concentration range of up to three log units. These results suggest the hypothesis that odorant representations in the turtle involve differential levels of input to many glomeruli, and that detecting relative patterns of distributed glomerular activation may be an important strategy for encoding odor quality independent of intensity.
Y5 Receptors Mediate Neuropeptide Y Actions at Excitatory Synapses in Area CA3 of the Mouse Hippocampus
Authors: Guo Hui , Castro Peter A. , Palmiter Richard D. , Baraban Scott C.
Published: May 2017

Neuropeptide Y (NPY) is a potent modulator of excitatory synaptic transmission and limbic seizures. NPY is abundantly expressed in the dentate gyrus and is thought to modulate hippocampal excitability via activation of presynaptic Y2 receptors (Y2R). Here we demonstrate that NPY, and commonly used Y2R-preferring (NPY13–36) and Y5 receptor (Y5R)–preferring ([d-Trp32]NPY and hPP) peptide agonists, evoke similar levels of inhibition at excitatory CA3 synapses in hippocampal slices from wild-type control mice (WT). In contrast, NPYergic inhibition of excitatory CA3 synaptic transmission is absent in mice lacking the Y5R subtype (Y5R KO). In both analyses of evoked population spike activity and spontaneous excitatory postsynaptic synaptic currents (EPSCs), NPY agonists induced powerful inhibitory effects in all hippocampal slices from WT mice, whereas these peptides had no effect in slices from Y5R KO mice. In slices from WT mice, NPY (and NPY receptor–preferring agonists) reduced the frequency of spontaneous EPSCs but had no effect on sEPSC amplitude, rise time, or decay time. Furthermore, NPYergic modulation of spontaneous EPSCs in WT mice was mimicked by bath application of a novel Y5R-selective peptide agonist ([cpp]hPP) but not the selective Y2R agonist ([ahx5–24]NPY). In situ hybridization was used to confirm the presence of NPY, Y2, and Y5 mRNA in the hippocampus of WT mice and the absence of Y5R in knockout mice. These results suggest that the Y5 receptor subtype, previously believed to mediate food intake, plays a critical role in modulation of hippocampal excitatory transmission at the hilar-to-CA3 synapse in the mouse.
Activation of Spinobulbar Lamina I Neurons by Static Muscle Contraction
Authors: Wilson L. B. , Andrew D. , Craig A. D.
Published: May 2017

Spinal lamina I neurons are selectively activated by small-diameter somatic afferents, and they project to brain stem sites that are critical for homeostatic control. Because small-diameter afferent activity evoked by contraction of skeletal muscle reflexly elicits exercise-related cardiorespiratory activation, we tested whether spinobulbar lamina I cells respond to muscle contraction. Spinobulbar lamina I neurons were identified in chloralose-anesthetized cats by antidromic activation from the ipsilateral caudal ventrolateral medulla. Static contractions of the ipsilateral triceps surae muscle were evoked by tibial nerve stimulation using parameters that avoid afferent activation, and arterial blood pressure responses were recorded. Recordings were maintained from 13 of 17 L7 lamina I spinobulbar neurons during static muscle contraction, and 5 of these neurons were excited. Three were selectively activated only by muscle afferents and did not have a cutaneous receptive field. Spinobulbar lamina I neurons activated by muscle contraction provide an ascending link for the reflex cardiorespiratory adjustments that accompany muscular work. This study provides an important first step in elucidating an ascending afferent pathway for somato-autonomic reflexes.
Human Gaze Stabilization During Active Head Translations
Authors: Medendorp W. P. , Van Gisbergen J.A.M. , Gielen C.C.A.M.
Published: May 2017

This study investigated how binocular gaze is controlled to compensate for self-generated translational movements of the head where geometric requirements dictate that the ideal gaze signal needs to be modulated by the inverse of target distance. Binocular gaze (eye plus head) was measured for visual and remembered targets at various distances in six human subjects during active head translations at frequencies of 0.25, 0.5, 1.0, and 1.5 Hz. We found that, during head translations, gaze changes were achieved by a combination of eye and head rotations. Accordingly, stabilization performance was characterized by the gaze response parameters sensitivity and phase, where sensitivity is defined as the ratio of gaze velocity and translational eye velocity and where phase refers to the phase delay of gaze velocity relative to translational eye velocity. In the analysis, we used a binocular coordinate system yielding a version and a vergence component. We examined how frequency and target distance, estimated from the vergence angle, affected sensitivity and phase of the version component of the gaze signal and compared the results to the requirements for ideal performance. The relation between gaze sensitivity and the inverse of distance was characterized by a linear regression analysis. The ratio of the slope of the linear regression and the slope required for ideal stabilization provided a measure for the degree of “distance compensation.” The results show that distance compensation was better for a visual target than for remembered targets in darkness, and behaved according to low-pass characteristics in both target conditions. It declined from 1.00 to 0.84 for visual targets and from 0.87 to 0.57 for remembered targets in the frequency range 0.25–1.5 Hz. The intercept obtained from the regression yielded the gaze response at zero vergence and specified a “default sensitivity” of gaze compensation. Default sensitivity increased with frequency from 0.02 at 0.25 Hz to 0.10°/cm at 1.5 Hz for visual targets and from 0.04 to 0.16°/cm in darkness. The phase delays of the gaze response increased on average from 2 to 7° in the frequency range 0.25–1.5 Hz. In comparison with earlier passive studies, active translation compensation in the dark is superior at all frequencies where comparison was possible. We conclude that a nonvestibular signal with low-pass characteristics contributes to gaze during active head translations.
Spinal Projections of the Cat Parvicellular Red Nucleus
Authors: Pong Milton , Horn Kris M. , Gibson Alan R.
Published: May 2017

Traditionally, the red nucleus of the cat is divided into two parts: a large-celled, magnocellular, division (RNm) and a small-celled, parvicellular, division (RNp). The RNm projects to the spinal cord and receives input from the cerebellar interpositus nucleus. The RNp projects to the inferior olive and receives input from the cerebellar dentate nucleus. In this report, we reexamine the connections of the red nucleus using the bidirectional tracer wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Our findings demonstrate that the cat RNp has a large caudal and lateral region that projects to contralateral spinal cord and not to the inferior olive. The spinally projecting region of RNp receives input from the dentate nucleus and a lateral segment of anterior interpositus. Cervical projections from the red nucleus show a topography with the rostral portion of RNp favoring upper segments and the caudal portion of RNm favoring lower segments. The results show that dentate output can influence spinal activity without passing through the cerebral cortex. For the control of movements such as reaching and grasping, we suggest that RNp and dentate focus on the control of proximal limb musculature, whereas RNm and the anterior interpositus focus on the control of distal limb musculature. We also suggest that other species are likely to have a small-celled area of red nucleus projecting to the spinal cord.
Inhibition of Mitochondrial Matrix Chaperones and Antiapoptotic Bcl-2 Family Proteins Empower Antitumor Therapeutic Responses
Authors: Karpel-Massler Georg , Ishida Chiaki Tsuge , Bianchetti Elena , Shu Chang , Perez-Lorenzo Rolando , Horst Basil , Banu Matei , Roth Kevin A. , Bruce Jeffrey N. , Canoll Peter , Altieri Dario C. , Siegelin Markus D.
Published: May 2017

Rational therapeutic approaches based on synthetic lethality may improve cancer management. On the basis of a high-throughput drug screen, we provide preclinical proof of concept that targeting the mitochondrial Hsp90 chaperone network (mtHsp90) and inhibition of Bcl-2, Bcl-xL, and Mcl-1 is sufficient to elicit synthetic lethality in tumors recalcitrant to therapy. Our analyses focused on BH3 mimetics that are broad acting (ABT263 and obatoclax) or selective (ABT199, WEHI-539, and A1210477), along with the established mitochondrial matrix chaperone inhibitor gamitrinib-TPP. Drug combinations were tested in various therapy-resistant tumors in vitro and in vivo in murine model systems of melanoma, triple-negative breast cancer, and patient-derived orthotopic xenografts (PDX) of human glioblastoma. We found that combining BH3 mimetics and gamitrinib-TPP blunted cellular proliferation in a synergistic manner by massive activation of intrinsic apoptosis. In like manner, suppressing either Bcl-2, Bcl-xL, or Mcl-1 recapitulated the effects of BH3 mimetics and enhanced the effects of gamitrinib-TPP. Mechanistic investigations revealed that gamitrinib-TPP activated a PERK-dependent integrated stress response, which activated the proapoptotic BH3 protein Noxa and its downstream targets Usp9X and Mcl-1. Notably, in the PDX glioblastoma and BRAFi-resistant melanoma models, this drug combination safely and significantly extended host survival. Our results show how combining mitochondrial chaperone and Bcl-2 family inhibitors can synergize to safely degrade the growth of tumors recalcitrant to other treatments. Cancer Res; 77(13); 3513–26. ©2017 AACR.
Preclinical Evaluation of MEDI0641, a Pyrrolobenzodiazepine-Conjugated Antibody–Drug Conjugate Targeting 5T4
Authors: Harper Jay , Lloyd Christopher , Dimasi Nazzareno , Toader Dorin , Marwood Rose , Lewis Leeanne , Bannister David , Jovanovic Jelena , Fleming Ryan , D'Hooge Francois , Mao Shenlan , Marrero Allison M. , Korade Martin , Strout Patrick , Xu Linda , Chen Cui , Wetzel Leslie , Breen Shannon , van Vlerken-Ysla Lilian , Jalla Sanjoo , Rebelatto Marlon , Zhong Haihong , Hurt Elaine M. , Hinrichs Mary Jane , Huang Keven , Howard Philip W. , Tice David A. , Hollingsworth Robert E. , Herbst Ronald , Kamal Adeela
Published: May 2017

Antibody–drug conjugates (ADC) are used to selectively deliver cytotoxic agents to tumors and have the potential for increased clinical benefit to cancer patients. 5T4 is an oncofetal antigen overexpressed on the cell surface in many carcinomas on both bulk tumor cells as well as cancer stem cells (CSC), has very limited normal tissue expression, and can internalize when bound by an antibody. An anti-5T4 antibody was identified and optimized for efficient binding and internalization in a target-specific manner, and engineered cysteines were incorporated into the molecule for site-specific conjugation. ADCs targeting 5T4 were constructed by site-specifically conjugating the antibody with payloads that possess different mechanisms of action, either a DNA cross-linking pyrrolobenzodiazepine (PBD) dimer or a microtubule-destabilizing tubulysin, so that each ADC had a drug:antibody ratio of 2. The resulting ADCs demonstrated significant target-dependent activity in vitro and in vivo; however, the ADC conjugated with a PBD payload (5T4-PBD) elicited more durable antitumor responses in vivo than the tubulysin conjugate in xenograft models. Likewise, the 5T4-PBD more potently inhibited the growth of 5T4-positive CSCs in vivo, which likely contributed to its superior antitumor activity. Given that the 5T4-PBD possessed both potent antitumor activity as well as anti-CSC activity, and thus could potentially target bulk tumor cells and CSCs in target-positive indications, it was further evaluated in non-GLP rat toxicology studies that demonstrated excellent in vivo stability with an acceptable safety profile. Taken together, these preclinical data support further development of 5T4-PBD, also known as MEDI0641, against 5T4+ cancer indications. Mol Cancer Ther; 16(8); 1576–87. ©2017 AACR.
Identification of Cancer-Targeted Tropomyosin Inhibitors and Their Synergy with Microtubule Drugs
Authors: Currier Mark A. , Stehn Justine R. , Swain Ashleigh , Chen Duo , Hook Jeff , Eiffe Eleanor , Heaton Andrew , Brown David , Nartker Brooke A. , Eaves David W. , Kloss Nina , Treutlein Herbert , Zeng Jun , Alieva Irina B. , Dugina Vera B. , Hardeman Edna C. , Gunning Peter W. , Cripe Timothy P.
Published: May 2017

Actin filaments, with their associated tropomyosin polymers, and microtubules are dynamic cytoskeletal systems regulating numerous cell functions. While antimicrotubule drugs are well-established, antiactin drugs have been more elusive. We previously targeted actin in cancer cells by inhibiting the function of a tropomyosin isoform enriched in cancer cells, Tpm3.1, using a first-in-class compound, TR100. Here, we screened over 200 other antitropomyosin analogues for anticancer and on-target activity using a series of in vitro cell-based and biochemical assays. ATM-3507 was selected as the new lead based on its ability to disable Tpm3.1-containing filaments, its cytotoxicity potency, and more favorable drug-like characteristics. We tested ATM-3507 and TR100 alone and in combination with antimicrotubule agents against neuroblastoma models in vitro and in vivo. Both ATM-3507 and TR100 showed a high degree of synergy in vitro with vinca alkaloid and taxane antimicrotubule agents. In vivo, combination-treated animals bearing human neuroblastoma xenografts treated with antitropomyosin combined with vincristine showed minimal weight loss, a significant and profound regression of tumor growth and improved survival compared with control and either drug alone. Antitropomyosin combined with vincristine resulted in G2–M phase arrest, disruption of mitotic spindle formation, and cellular apoptosis. Our data suggest that small molecules targeting the actin cytoskeleton via tropomyosin sensitize cancer cells to antimicrotubule agents and are tolerated together in vivo. This combination warrants further study. Mol Cancer Ther; 16(8); 1555–65. ©2017 AACR.
Sensitization of EGFR Wild-Type Non–Small Cell Lung Cancer Cells to EGFR-Tyrosine Kinase Inhibitor Erlotinib
Authors: Raimbourg Judith , Joalland Marie-Pierre , Cabart Mathilde , de Plater Ludmilla , Bouquet Fanny , Savina Ariel , Decaudin Didier , Bennouna Jaafar , Vallette François M. , Lalier Lisenn
Published: May 2017

The benefit of EGFR–TKI in non–small cell lung cancer has been demonstrated in mutant EGFR tumors as first-line treatment but the benefit in wild-type EGFR tumors is marginal as well as restricted to maintenance therapy in pretreated patients. This work aimed at questioning the effects of cisplatin initial treatment on the EGFR pathway in non–small cell lung cancer and the functional consequences in vitro and in in vivo animal models of patient-derived xenografts (PDX). We establish here that cisplatin pretreatment specifically sensitizes wild-type EGFR-expressing cells to erlotinib, contrary to what happens in mutant EGFR cells and with a blocking EGFR antibody, both in vitro and in vivo. The sensitization entails the activation of the kinase Src upstream of EGFR, thereafter transactivating EGFR through a ligand-independent activation. We propose a combination of markers that enable to discriminate between the tumors sensitized to erlotinib or not in PDX models, which should be worth testing in patients. These markers might be useful for the selection of patients who would benefit from erlotinib as a maintenance therapy. Mol Cancer Ther; 16(8); 1634–44. ©2017 AACR.
Soluble PD-L1 as a Biomarker in Malignant Melanoma Treated with Checkpoint Blockade
Authors: Zhou Jun , Mahoney Kathleen M. , Giobbie-Hurder Anita , Zhao Fengmin , Lee Sandra , Liao Xiaoyun , Rodig Scott , Li Jingjing , Wu Xinqi , Butterfield Lisa H. , Piesche Matthias , Manos Michael P. , Eastman Lauren M. , Dranoff Glenn , Freeman Gordon J. , Hodi F. Stephen
Published: May 2017

Blockade of the pathway including programmed death-ligand 1 (PD-L1) and its receptor programmed cell death protein 1 (PD-1) has produced clinical benefits in patients with a variety of cancers. Elevated levels of soluble PD-L1 (sPD-L1) have been associated with worse prognosis in renal cell carcinoma and multiple myeloma. However, the regulatory roles and function of sPD-L1 particularly in connection with immune checkpoint blockade treatment are not fully understood. We identified four splice variants of PD-L1 in melanoma cells, and all of them are secreted. Secretion of sPD-L1 resulted from alternate splicing activities, cytokine induction, cell stress, cell injury, and cell death in melanoma cells. Pretreatment levels of sPD-L1 were elevated in stage IV melanoma patient sera compared with healthy donors. High pretreatment levels of sPD-L1 were associated with increased likelihood of progressive disease in patients treated by CTLA-4 or PD-1 blockade. Although changes in circulating sPD-L1 early after treatment could not distinguish responders from those with progressive disease, after five months of treatment by CTLA-4 or PD-1 blockade patients who had increased circulating sPD-L1 had greater likelihood of developing a partial response. Induction of sPD-L1 was associated with increased circulating cytokines after CTLA-4 blockade but not following PD-1 blockade. Circulating sPD-L1 is a prognostic biomarker that may predict outcomes for subgroups of patients receiving checkpoint inhibitors. Cancer Immunol Res; 5(6); 480–92. ©2017 AACR.