bluuebunny/crossref_metadata_2025 huggingface dataset:
Temporal dynamics of pulmonary response to intravenous histamine in dogs: effects of dose and lung volume
Authors: Bates J. H. , Lauzon A. M. , Dechman G. S. , Maksym G. N. , Schuessler T. F.
Published: December 2017

We measured tracheal pressure (Ptr) and tracheal flow (V) in open-chest anesthetized paralyzed dogs. The lungs were maintained at a fixed volume (initial positive end-expiratory pressure 0.5 kPa) for 80 s while small-amplitude oscillations in V at 1 and 6 Hz were applied simultaneously at the tracheal opening. A bolus of histamine was given intravenously at the start of the oscillation period. The time course of lung elastic recoil pressure (Pel) was obtained by passing a running average over Ptr to smooth out its oscillations. The oscillations themselves were separated into their 1- and 6-Hz components, as were those in V. By fitting models to the 1- and 6-Hz components of Ptr and V by recursive least squares, we obtained time courses of lung resistance at 6 Hz (RL6), dynamic lung elastance at 1 Hz (EL1), and the difference between dynamic lung resistance at 1 and 6 Hz (RL1-RL6). In four dogs we studied the effects of histamine doses of 0.05, 1.0, and 20 mg. We found that Pel increased quickly and plateaued, RL6 continued to increase throughout the oscillation period, and EL1 exhibited features of both Pel and RL6. Furthermore, the ratio of RL1-RL6 to EL1 was qualitatively similar in time course to Pel. We explain these varied time courses in terms of the development of regional ventilation inhomogeneity throughout the lung as the reaction to histamine develops. In four dogs we also studied the effects of reducing the initial positive end-expiratory pressure by 0.25 kPa and found that the changes in RL6, EL1, and RL1-RL6 were greatly magnified, presumably because of the reduced forces of parenchymal interdependence.
Hering-Breuer reflex and respiratory system compliance in the first year of life: a longitudinal study
Authors: Rabbette P. S. , Fletcher M. E. , Dezateux C. A. , Soriano-Brucher H. , Stocks J.
Published: December 2017

The airway occlusion technique for measuring passive respiratory mechanics in infants relies on an ability to evoke the Hering-Breuer lung inflation reflex (HBR). However, there is conflicting evidence regarding the persistence of the HBR beyond the early newborn period. This study was designed to assess maturational changes in HBR activity and passive total respiratory system compliance (Crs) in healthy infants during the 1st yr of life. The strength of the HBR was assessed from the relative change in expiratory time (TE) after brief end-inspiratory airway occlusions compared with resting TE during spontaneous breathing. Crs was measured using the multiple-occlusion technique. Paired measurements of HBR activity and Crs were obtained during sedated sleep in 30 infants at 4–8 wk and at 1 yr of age. Significant HBR activity during tidal breathing persisted throughout the 1st yr of life, with TE increasing during occlusion by at least 26% in every infant. However, the relative strength of the reflex response decreased from a mean of 88.3% (range, 34–160%) at approximately 6 wk to 50.3% (range, 26–125%) by 1 yr of age (P < 0.001). All infants showed an increase in Crs with age, with mean Crs increasing from 60.1 +/- 8.9 (SD) to 149.0 +/- 20.6 ml/kPa between 6 wk and 1 yr. However, there was no apparent relationship between the magnitude of decline in HBR response and the age-related changes in Crs.(ABSTRACT TRUNCATED AT 250 WORDS)
Inhibition of aldosterone release by hypoxia in vitro: interaction with carbon monoxide
Authors: Raff H. , Jankowski B.
Published: December 2017

We have demonstrated that the aldosteronogenic pathway of the zona glomerulosa is unusually sensitive to modest changes in PO2 (Michaelis constant for O2 approximately 95 Torr). The current study evaluated the interaction of CO (the classic ligand for P-450 enzymes) and the decreases in O2 on aldosteronogenesis in vitro. Bovine adrenocortical zona glomerulosa cells were incubated for 2 h and stimulated with either adenosine 3′,5′-cyclic monophosphate (cAMP) or angiotensin II. Ten and 20% CO led to significant decreases in cAMP- and angiotensin II-stimulated aldosteronogenesis. The combination of 20% CO and moderate decreases in PO2 (from approximately 140 to approximately 100 Torr) led to an interactive decrease in aldosterone production. The conversion of corticosterone to aldosterone catalyzed by aldosterone synthase, which is the site of O2 sensitivity, was not significantly inhibited by CO. We conclude that the aldosterone pathway is not exceptionally sensitive to CO compared with other steroidogenic pathways. This observation suggests that the unique O2-sensitive properties of the aldosterone pathway located primarily within aldosterone synthase may not reside in its CO binding site (i.e., heme).
Retrotrapezoid nucleus glutamate injections: long-term stimulation of phrenic activity
Authors: Nattie E. E. , Li A.
Published: December 2017

In chloralose-urethan anesthetized, paralyzed, vagotomized, glomectomized, and servo-ventilated cats we examined the effects of 10 nl of glutamate (10 mM, 100 mM, and 1 M) injected unilaterally over 60 s into the region of the retrotrapezoid nucleus (RTN). Seven 10 mM glutamate injections produced no consistent effects on the amplitude of the integrated phrenic nerve signal, respiratory cycle duration, or blood pressure. Ten 100 mM injections consistently increased integrated phrenic amplitude significantly from a baseline average of 31 +/- 2% (SE) of maximum to a peak response average of 50 +/- 3% of maximum. This effect was long lasting (45.6 +/- 8.6 min). Blood pressure responses were variable. Seven 1 M glutamate injections consistently decreased integrated phrenic amplitude significantly from a baseline average for all injections of 29 +/- 3% of maximum to a peak average of 20 +/- 5% of maximum. Respiratory cycle duration and blood pressure responses were variable. Prior injection into the RTN of 10 nl of 100 mM kynurenic acid attenuated the subsequent response of the integrated phrenic amplitude to injection of 10 nl of glutamate at the same site. Comparison of glutamate (10 nl, 100 mM) injected over 60 s vs. 30 ms showed that the prolonged increase in phrenic activity was related to the longer-duration (60-s) injections and that RTN single units were stimulated for up to 5 min on average after the 60-s injection with one unit showing an increase in firing rate over 40 min. After the 30-ms injection, integrated phrenic amplitude and RTN unit mean firing rate were increased for the first two breaths and at 5 min after the injection. We conclude that glutamate injected into the RTN increases local single-unit firing rate and the amplitude of the integrated phrenic activity. Injections made over 60 s result in prolonged phrenic stimulation and, in some cases, in RTN single-unit firing rate.
Physiological Evidence for Ionotropic and Metabotropic Glutamate Receptors in Rat Taste Cells
Authors: Lin Weihong , Kinnamon Sue C.
Published: December 2017

Monosodium glutamate (MSG) elicits a unique taste in humans called umami. Recent molecular studies suggest that glutamate receptors similar to those in brain are present in taste cells, but their precise role in taste transduction remains to be elucidated. We used giga-seal whole cell recording to examine the effects of MSG and glutamate receptor agonists on membrane properties of taste cells from rat fungiform papillae. MSG (1 mM) induced three subsets of responses in cells voltage-clamped at −80 mV: a decrease in holding current (subset I), an increase in holding current (subset II), and a biphasic response consisting of an increase, followed by a decrease in holding current (subset III). Most subset II glutamate responses were mimicked by the ionotropic glutamate receptor (iGluR) agonist N-methyl-d-aspartate (NMDA). The current was potentiated by glycine and was suppressed by the NMDA receptor antagonist d(−)-2-amino-5-phosphonopentanoic acid (AP5). The group III metabotropic glutamate receptor (mGluR) agonistl-2-amino-4-phosphonobutyric acid (l-AP4) usually mimicked the subset I glutamate response. This hyperpolarizing response was suppressed by the mGluR antagonist (RS)-α-cyclopropyl-4-phosphonophenylglycine (CPPG) and by 8-bromo-cAMP, suggesting a role for cAMP in the transduction pathway. In a small subset of taste cells, l-AP4 elicited an increase in holding current, resulting in taste cell depolarization under current clamp. Taken together, our results suggest that NMDA-like receptors and at least two types of group III mGluRs are present in taste receptor cells, and these may be coactivated by MSG. Further studies are required to determine which receptors are located on the apical membrane and how they contribute to the umami taste.
Two Types of Voltage-Gated K+Currents in Dissociated Heart Ventricular Muscle Cells of the SnailLymnaea stagnalis
Authors: Yeoman M. S. , Benjamin P. R.
Published: December 2017

We have used a combination of current-clamp and voltage-clamp techniques to characterize the electrophysiological properties of enzymatically dissociated Lymnaea heart ventricle cells. Dissociated ventricular muscle cells had average resting membrane potentials of −55 ± 5 mV. When hyperpolarized to potentials between −70 and −63 mV, ventricle cells were capable of firing repetitive action potentials (8.5 ± 1.2 spikes/min) that failed to overshoot 0 mV. The action potentials were either simple spikes or more complex spike/plateau events. The latter were always accompanied by strong contractions of the muscle cell. The waveform of the action potentials were shown to be dependent on the presence of extracellular Ca2+and K+ions. With the use of the single-electrode voltage-clamp technique, two types of voltage-gated K+currents were identified that could be separated by differences in their voltage sensitivity and time-dependent kinetics. The first current activated between −50 and −40 mV. It was relatively fast to activate (time-to-peak; 13.7 ± 0.7 ms at +40 mV) and inactivated by 53.3 ± 4.9% during a maintained 200-ms depolarization. It was fully available for activation below −80 mV and was completely inactivated by holding potentials more positive than −40 mV. It was completely blocked by 5 mM 4-aminopyridine (4-AP) and by concentrations of tetraethylammonium chloride (TEA) >10 mM. These properties characterize this current as a member of the A-type family of voltage-dependent K+currents. The second voltage-gated K+current activated at more depolarized potentials (−30 to −20 mV). It activated slower than the A-type current (time-to-peak; 74.1 ± 3.9 ms at +40 mV) and showed little inactivation (6.2 ± 2.1%) during a maintained 200-ms depolarization. The current was fully available for activation below −80 mV with a proportion of the current still available for activation at potentials as positive as 0 mV. The current was completely blocked by 1–3 mM TEA. These properties characterize this current as a member of the delayed rectifier family of voltage-dependent K+currents. The slow activation rates and relatively depolarized activation thresholds of the two K+currents are suggestive that their main role is to contribute to the repolarization phase of the action potential.
Peripheral Coding of Tonic Mechanical Cutaneous Pain: Comparison of Nociceptor Activity in Rat and Human Psychophysics
Authors: Andrew David , Greenspan Joel D.
Published: December 2017

These experiments investigated temporal summation mechanisms of tonic cutaneous mechanical pain. Human volunteers provided psychophysical estimates of pain intensity, which were compared with discharge patterns of rat cutaneous nociceptors tested with identical stimulus protocols. Human subjects made either intermittent or continuous ratings of pain intensity during stimulation of the skin between the thumb and first finger. Stimulus intensities of 25, 50, and 100 g were applied with a probe of contact area of 0.1 mm2 for 2 min. Pain perception significantly increased during stimulation (temporal summation) for the 50- and 100-g stimulus intensities. Sequential conduction block of the myelinated fibers supplying the stimulated skin was used to investigate the role of A-fiber mechanoreceptors and nociceptors in this temporal summation. Conduction block of the Aβ fibers resulted in an increase in mechanically evoked pain estimates and an increase in temporal summation, consistent with loss of Aβ-mediated inhibition. When only conduction in the unmyelinated fibers remained, pain estimates were reduced to the preblock levels, but temporal summation was still present. Electrophysiological recordings were made from filaments of the sciatic nerve supplying receptors in the plantar skin of barbiturate-anesthetized rats. Forty units fulfilled the identification criteria for nociceptors: 20 A-fiber and 20 C-fiber nociceptors. Each unit was characterized by recording its responses to graded mechanical and heat stimuli. Nociceptors were also tested with stimuli identical to those applied to the human subjects. The responses of all units to sustained mechanical stimuli were adaptive—that is, they exhibited a gradual decline in response with time. However, the time course of adaptation varied among units. All the C-fiber nociceptors and one-half of the A-fiber nociceptors had rapidly adapting responses. The remainder of the A-fibers displayed slowly adapting responses. One-third of all units also showed short-duration increases in firing rate during stimulation. The latency after stimulus onset of this rate acceleration was inversely related to stimulus intensity. Despite the apparent disparity between perceptual temporal summation and nociceptor adaptation, central and peripheral mechanisms are proposed that can reconcile the relationship between nociceptor activity and pain perception.
Apparent Dissociation Between Saccadic Eye Movements and the Firing Patterns of Premotor Neurons and Motoneurons
Authors: Ling Leo , Fuchs Albert F. , Phillips James O. , Freedman Edward G.
Published: December 2017

Saccadic eye movements result from high-frequency bursts of activity in ocular motoneurons. This phasic activity originates in premotor burst neurons. When the head is restrained, the number of action potentials in the bursts of burst neurons and motoneurons increases linearly with eye movement amplitude. However, when the head is unrestrained, the number of action potentials now increase as a function of the change in the direction of the line of sight during eye movements of relatively similar amplitudes. These data suggest an apparent uncoupling of premotor neuron and motoneuron activity from the resultant eye movement.
Apamin-Sensitive Conductance Mediates the K+ Current Response During Chemical Ischemia in CA3 Pyramidal Cells
Authors: Tanabe Mitsuo , Mori Masahiro , Gähwiler Beat H. , Gerber Urs
Published: December 2017

Pyramidal cells typically respond to ischemia with initial transient hyperpolarization, which may represent a neuroprotective response. To identify the conductance underlying this hyperpolarization in CA3 pyramidal neurons of rat hippocampal organotypic slice cultures, recordings were obtained using the single-electrode voltage-clamp technique. Brief chemical ischemia (2 mM 2-deoxyglucose and 3 mM NaN3, for 4 min) induced a response mediated by an increase in K+ conductance. This current was blocked by intracellular application of the Ca2+ chelator, bis-( o-aminophenoxy)- N,N,N′,N′-tetraacetic acid (BAPTA), reduced with low external [Ca2+], and inhibited by a selective L-type Ca2+ channel inhibitor, isradipine, consistent with the activation of a Ca2+-dependent K+ conductance. Experiments with charybdotoxin (10 nM) and tetraethylammonium (TEA; 1 mM), or with the protein kinase C activator, phorbol 12,13-diacetate (PDAc; 3 μM), ruled out an involvement of a large conductance–type or an apamin-insensitive small conductance, respectively. In the presence of apamin (1 μM), however, the outward current was significantly reduced. These results demonstrate that in rat hippocampal CA3 pyramidal neurons an apamin-sensitive Ca2+-dependent K+ conductance is activated in response to brief ischemia generating a pronounced outward current.
Activity-Related Calcium Dynamics in Motoneurons of the Nucleus Hypoglossus From Mouse
Authors: Lips Mario B. , Keller Bernhard U.
Published: December 2017

A quantitative analysis of activity-related calcium dynamics was performed in motoneurons of the nucleus hypoglossus in the brain stem slice preparation from mouse by simultaneous patch-clamp and microfluorometric calcium measurements. Motoneurons were analyzed under in vitro conditions that kept them in a functionally intact state represented by rhythmic, inspiratory-related bursts of excitatory postsynaptic currents and associated action potential discharges. Bursts of electrical activity were paralleled by somatic calcium transients resulting from calcium influx through voltage-activated calcium channels, where each action potential accounted for a calcium-mediated charge influx around 2 pC into the somatic compartment. Under in vivo conditions, rhythmic-respiratory activity in young mice occurred at frequencies up to 5 Hz, demonstrating the necessity for rapid calcium elevation and recovery in respiratory-related neurons. The quantitative analysis of hypoglossal calcium homeostasis identified an average extrusion rate, but an exceptionally low endogenous calcium binding capacity as cellular parameters accounting for rapid calcium signaling. Our results suggest that dynamics of somatic calcium transients 1) define an upper limit for the maximum frequency of respiratory-related burst discharges and 2) represent a potentially dangerous determinant of intracellular calcium profiles during pathophysiological and/or excitotoxic conditions.
Visual Stimulus–Dependent Changes in Interhemispheric EEG Coherence in Ferrets
Authors: Kiper D. C. , Knyazeva M. G. , Tettoni L. , Innocenti G. M.
Published: December 2017

In recent years, the analysis of the coherence between signals recorded from the scalp [electroencephalographic (EEG) coherence] has been used to assess the functional properties of cortico-cortical connections, both in animal models and in humans. However, the experimental validation of this technique is still scarce. Therefore we applied it to the study of the callosal connections between the visual areas of the two hemispheres, because this particular set of cortico-cortical connections can be activated in a selective way by visual stimuli. Indeed, in primary and in low-order secondary visual areas, callosal axons interconnect selectively regions, which represent a narrow portion of the visual field straddling the vertical meridian and, within these regions, neurons that prefer the same stimulus orientation. Thus only isooriented stimuli located near the vertical meridian are expected to change interhemispheric coherence by activating callosal connections. Finally, if such changes are found and are indeed mediated by callosal connections, they should disappear after transection of the corpus callosum. We perfomed experiments on seven paralyzed and anesthetized ferrets, recording their cortical activity with epidural electrodes on areas 17/18, 19, and lateral suprasylvian, during different forms of visual stimulation. As expected, we found that bilateral iso-oriented stimuli near the vertical meridian, or extending across it, caused a significant increase in interhemispheric coherence in the EEG beta-gamma band. Stimuli with different orientations, stimuli located far from the vertical meridian, as well as unilateral stimuli failed to affect interhemispheric EEG coherence. The stimulus-induced increase in coherence disappeared after surgical transection of the corpus callosum. The results suggest that the activation of cortico-cortical connections can indeed be revealed as a change in EEG coherence. The latter can therefore be validly used to investigate the functionality of cortico-cortical connections.
Spontaneous and Artificial Activation of Neocortical Seizures
Authors: Amzica Florin , Steriade Mircea
Published: December 2017

The aim of this study is to disclose the mechanisms underlying the recruitment of neocortical networks during slow-wave sleep oscillations evolving into spike-wave (SW) seizures. 1) We investigated the activation of SW seizures in a seizure-prone neocortex by means of electrical stimuli applied within the frequency range of spontaneous sleep oscillations. Stimuli were grouped in bursts of 10 Hz, similar to sleep spindles, and repeated every 2 s, to reproduce their rhythmic recurrence imposed by the slow (<1 Hz) sleep oscillation. Either cortical or thalamic stimuli, which were applied while the cortex displayed sleeplike activity, gradually induced paroxysmal responses in intracellularly recorded neocortical neurons, which were virtually identical to those of spontaneous seizures and consisted of a progressive buildup of paroxysmal depolarizing shifts (PDSs). 2) The ability of cortical networks to follow stimuli was tested at various stimulation frequencies (1–3 Hz) and quantified by calculating the entropy of the ensuing oscillation. Rhythmic PDSs were optimally induced, and the lowest entropy was generated, at a stimulation frequency around 1.5 Hz. Fast runs at 10–15 Hz, which often override PDSs, thus contributing to the polyspike-wave pattern of seizures, were induced by cortical stimuli, but were disturbed by thalamic stimuli. Spontaneous seizures generally evolved toward an accelerated discharge of PDSs. It is suggested that these accelerating trends during SW seizures act as protective mechanisms by provoking the uncoupling of cortical networks and eventually arresting the seizure.
Gentamicin Blocks Both Fast and Slow Effects of Olivocochlear Activation in Anesthetized Guinea Pigs
Authors: Yoshida Naohiro , Liberman M. Charles , Brown M. Christian , Sewell William F.
Published: December 2017

The medial olivocochlear (MOC) efferent system, which innervates cochlear outer hair cells, suppresses cochlear responses. MOC-mediated suppression includes both slow and fast components, with time courses differing by three orders of magnitude. Pharmacological studies in anesthetized guinea pigs suggest that both slow and fast effects on cochlear responses require an initial acetylcholine activation of α-9 nicotinic receptors on outer hair cells and that slow effects require additional intracellular events downstream from those mediating fast effects. Gentamicin, an aminoglycoside antibiotic, has been reported to block fast effects of sound-evoked OC activation following intramuscular injection in unanesthetized guinea pigs, without changing slow effects. In the present study, we show that electrically evoked fast and slow effects in the anesthetized guinea pig are both blocked by either intramuscular or intracochlear gentamicin, with similar time courses and/or dose-response curves. We suggest that sound-evoked slow effects in unanesthetized animals are fundamentally different from electrically evoked slow effects in anesthetized animals, and that the former may arise from effects of the lateral OC system.
Cross Talk Between A1 and A2A Adenosine Receptors in the Hippocampus and Cortex of Young Adult and Old Rats
Authors: Lopes Luísa V. , Cunha Rodrigo A. , Ribeiro J. A.
Published: December 2017

Adenosine modulates synaptic transmission by acting on inhibitory A1 and facilitatory A2A receptors, the densities of which are modified in aged animals. We investigated how A2A receptor activation influences A1receptor function and whether this interaction is modified in aged rats. In hippocampal and cortical nerve terminals from young adult (6 wk), but not old rats (24 mo), the A2A receptor agonist, 2-[4-(2-carboxyethyl) phenethylamino]-5′- N-ethylcarboxamidoadenosine (CGS 21680; 30 nM) decreased the binding affinity of a selective A1 receptor agonist, cyclopentyladenosine (CPA), an effect prevented by the A2A antagonist, (4-(2-[7-amino-2-(2-furyl {1,2,4}-triazolo{2,3-a {1,3,5}triazin-5-yl-aminoethyl)phenol (ZM 241385, 20 nM). This effect of CGS 21680 required intact nerve terminals and was also observed in the absence of Ca2+. This A2A-induced “desensitization” of A1receptors was prevented by the protein kinase C inhibitor, chelerythrine (6 μM), and was not detected in the presence of the protein kinase C activator, phorbol-12,13-didecanoate (250 nM), which itself caused a reduction in binding affinity for CPA. The protein kinase A inhibitor, N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (10 μM), and the protein kinase A activator, 8-Br-cAMP (1 mM), had no effects on the A2A-induced A1 receptor desensitization. This A2A-induced A1 receptor desensitization had a functional correlation because CGS 21680 (10 nM) attenuated by 40% the inhibition caused by CPA (10 nM) on CA1 area population spike amplitude in hippocampal slices. This A2A/A1 interaction may explain the attenuation by adenosine deaminase (2 U/ml), which removes tonic A1inhibition, of the facilitatory effect of CGS 21680 on synaptic transmission. The requirement of tonic A1 receptor activation for CGS 21680 to induce facilitation of synaptic transmission was reinforced by the observation that the A1receptor antagonist, 1,3-dipropyl-8-cyclopentylxanthine (20 nM) prevented CGS 21680 (10 nM) facilitation of population spike amplitude. The present results show the ability of A2A receptors to control A1 receptor function in a manner mediated by protein kinase C, but not protein kinase A, in young adult but not in aged rats.
Limited Contributions of Serotonin to Long-Term Hyperexcitability of Aplysia Sensory Neurons
Authors: Liao Xiaogang , Brou Christine G. , Walters Edgar T.
Published: December 2017

Serotonin (5-HT) has provided a useful tool to study plasticity of nociceptive sensory neurons in Aplysia. Because noxious stimulation causes release of 5-HT and long-term hyperexcitability (LTH) of sensory neuron somata and because 5-HT treatment can induce long-term synaptic facilitation of sensory neuron synapses, a plausible hypothesis is that 5-HT also induces LTH of the sensory neuron soma. Prolonged or repeated exposure of excised ganglia to 5-HT produced immediate hyperexcitability of sensory neurons that showed little desensitization, but the hyperexcitability decayed within minutes of washing out the 5-HT. Prolonged or repeated treatment of either excised ganglia or dissociated sensory neurons with various concentrations of 5-HT failed to induce significant LTH even when long-term synaptic facilitation was induced in the same preparations. Use of a high-divalent cation solution to reduce interneuron activity during 5-HT treatment failed to enable the induction of LTH in excised ganglia. Pairing 5-HT application with nerve shock failed to enhance LTH produced by nerve shock or to reveal covert LTH produced by 5-HT. The induction of LTH by nerve stimulation was enhanced rather than inhibited by treatment with methiothepin, a 5-HT antagonist reported to block various 5-HT receptors and 5-HT-induced adenylyl cyclase activation. This suggests that endogenous 5-HT may have inhibitory effects on the induction of LTH by noxious stimulation. Methiothepin blocked immediate hyperexcitability produced by exogenous 5-HT and also inhibited the expression of LTH induced by nerve stimulation when applied during testing 1 day afterward. At higher concentrations, methiothepin reduced basal excitability of sensory neurons by mechanisms that may be independent of its antagonism of 5-HT receptors. Several observations suggest that early release of 5-HT and consequent cAMP synthesis in sensory neurons is not important for the induction of LTH by noxious stimulation, whereas later release of 5-HT from persistently activated modulatory neurons, with consequent elevation of cAMP synthesis, may contribute to the maintenance of LTH.
Mechanical and Thermal Hyperalgesia and Ectopic Neuronal Discharge After Chronic Compression of Dorsal Root Ganglia
Authors: Song Xue-Jun , Hu San-Jue , Greenquist Kenneth W. , Zhang Jun-Ming , LaMotte Robert H.
Published: December 2017

Chronic compression of the dorsal root ganglion (CCD) was produced in adult rats by implanting a stainless steel rod unilaterally into the intervertebral foramen, one rod at L4 and another at L5. Two additional groups of rats received either a sham surgery or an acute injury consisting of a transient compression of the ganglion. Withdrawal of the hindpaw was used as evidence of a nocifensive response to mechanical and thermal stimulation of the plantar surface. In addition, extracellular electrophysiological recordings of spontaneous discharges were obtained from dorsal root fibers of formerly compressed ganglia using an in vitro nerve-DRG-dorsal root preparation. The mean threshold force of punctate indentation and the mean threshold temperature of heating required to elicit a 50% incidence of foot withdrawal ipsilateral to the CCD were significantly lower than preoperative values throughout the 35 days of postoperative testing. The number of foot withdrawals ipsilateral to the CCD during a 20-min contact with a temperature-controlled floor was significantly increased over preoperative values throughout postoperative testing when the floor was 4°C (hyperalgesia) and, to a lesser extent, when it was 30°C (spontaneous pain). Stroking the foot with a cotton wisp never elicited a reflex withdrawal before surgery but did so in most rats tested ipsilateral to the CCD during the first 2 postoperative weeks. In contrast, the CCD produced no changes in responses to mechanical or thermal stimuli on the contralateral foot. The sham operation and acute injury produced no change in behavior other than slight, mechanical hyperalgesia for ∼1 day, ipsilateral to the acute injury. Ectopic spontaneous discharges generated within the chronically compressed ganglion and, occurring in the absence of blood-borne chemicals and without an intact sympathetic nervous system, were recorded from neurons with intact, conducting, myelinated or unmyelinated peripheral nerve fibers. The incidence of spontaneously active myelinated fibers was 8.61% for CCD rats versus 0.96% for previously nonsurgical rats. We hypothesize that a chronic compression of the dorsal root ganglion after certain injuries or diseases of the spine may produce, in neurons with intact axons, abnormal ectopic discharges that originate from the ganglion and potentially contribute to low back pain, sciatica, hyperalgesia, and tactile allodynia.
Enhanced Excitability of Sensory Neurons in Rats With Cutaneous Hyperalgesia Produced by Chronic Compression of the Dorsal Root Ganglion
Authors: Zhang Jun-Ming , Song Xue-Jun , LaMotte Robert H.
Published: December 2017

Pain and hyperalgesia can occur when the dorsal root ganglion (DRG) and its roots are deformed mechanically in association with injuries or diseases of the spine. To evaluate the electrophysiological changes that contribute to this sensory pathology, intracellular recordings were obtained in vitro from DRGs that had received a chronic mechanical compression [chronic compression of DRG (CCD)]. The compression was produced by inserting L-shaped rods ipsilaterally into the intervertebral foramina, one at L4and the other at L5 in rats 1–14 days before the recording. Control rats received a sham operation. Postoperatively, the threshold force applied by punctate stimulation of the plantar surface of the hind paw decreased significantly on the foot ipsilateral to the CCD (mechanical hyperalgesia) but changed little on the contralateral foot or on either foot for control rats. DRG somata were viewed through a microscope during recording and classified as small, medium, and large according to their diameters. CCD cells in each size category were more excitable than those of comparable size from control rats as judged by the significantly lowered threshold currents and action potential voltage thresholds. Spontaneous activity was recorded in 11% of all the CCD cells tested. The spontaneous activity and/or changes in both the threshold currents and action potential thresholds were observed as early as one day after injury. The association of cutaneous hyperalgesia with changes in the electrophysiological properties of DRG cells suggests a possible role for intrinsic alterations in the membrane properties of compressed DRG cells in the production and persistence of chronic pain after certain spinal injuries or pathologies of the spine.
Pattern Generation in the Buccal System of Freely BehavingLymnaea stagnalis
Authors: Jansen Rene F. , Pieneman Anton W. , Maat Andries ter
Published: December 2017

Central pattern generators (CPGs) are neuronal circuits that drive active repeated movements such as walking or swimming. Although CPGs are, by definition, active in isolated central nervous systems, sensory input is thought play an important role in adjusting the output of the CPGs to meet specific behavioral requirements of intact animals. We investigated, in freely behaving snails ( Lymnaea stagnalis), how the buccal CPG is used during two different behaviors, feeding and egg laying. Analysis of the relationship between unit activity recorded from buccal nerves and the movements of the buccal mass showed that electrical activity in laterobuccal/ventrobuccal (LB/VB) nerves was as predicted from in vitro data, but electrical activity in the posterior jugalis nerve was not. Autodensity and interval histograms showed that during feeding the CPG produces a much stronger rhythm than during egg laying. The phase relationship between electrical activity and buccal movement changed little between the two behaviors. Fitting the spike trains recorded during the two behaviors with a simple model revealed differences in the patterns of electrical activity produced by the buccal system during the two behaviors investigated. During egg laying the bursts contained less spikes, and the number of spikes per burst was significantly more variable than during feeding. The time between two bursts of in a spike train was longer during egg laying than during feeding. The data show what the qualitative and quantitative differences are between two motor patterns produced by the buccal system of freely behaving Lymnaea stagnalis.
Asymmetry of Hindlimb Muscle Activity and Cutaneous Reflexes After Tendon Transfers in Kittens
Authors: Loeb G. E.
Published: December 2017

The mechanical actions of various ankle muscles were changed by surgically crossing or transferring the tendons in kittens. After the kittens grew to adults, both hindlimbs were implanted with multiple electromyogram (EMG) recording and cutaneous nerve stimulation electrodes to compare the activity of altered and normal muscles. The tendon transfers showed a remarkable tendency to regrow toward normal or only slightly altered mechanical action. In these animals and in the sham-operation controls, the patterns of muscle activity and reflexes were symmetrical in corresponding muscles of the two legs, although they could differ substantially between animals, particularly for the cutaneous reflexes. Eleven animals had at least some persistent alterations in muscle action. Their cutaneous reflex patterns tended to be asymmetric, in some cases quite markedly. EMG activity during unperturbed locomotion and paw-shaking was more symmetrical, but there were some changes in altered muscles and their synergists. The central pattern generators for locomotion and paw-shaking and particularly for cutaneous reflexes during locomotion appear to be at least partially malleable rather than entirely hardwired. This may provide a tool for studying their development and spinal plasticity in general.
Role of α-SNAP in Promoting Efficient Neurotransmission at the Crayfish Neuromuscular Junction
Authors: He Ping , Southard R. Chase , Chen Dong , Whiteheart S. W. , Cooper R. L.
Published: December 2017

In this manuscript, we address the role of the soluble N-ethylmaleimide sensitive factor attachment protein (α-SNAP) in synaptic transmission at the neuromuscular junction of the crayfish opener muscle. Immunochemcial methods confirm the presence of α-SNAP in these preparations and show that it is concentrated in the synaptic areas. Microinjection and electrophysiological studies show that α-SNAP causes an increase in neurotransmitter release yet does not significantly affect the kinetics. More specific quantal analysis, using focal, macropatch, synaptic current recordings, shows that α-SNAP increases transmitter release by increasing the probability of exocytosis but not the number of potential release sites. These data demonstrate that the role of α-SNAP is to increase the efficiency of neurotransmission by increasing the probability that a stimulus will result in neurotransmitter release. What this suggests is that α-SNAP is critical for the formation and maintenance of a “ready release” pool of synaptic vesicles.
Differential Inhibition of Glial K+ Currents by 4-AP
Authors: Bordey Angélique , Sontheimer Harald
Published: December 2017

Spinal cord astrocytes express four biophysically and pharmacologically distinct voltage-activated potassium (K+) channel types. The K+ channel blocker 4-aminopyridine (4-AP) exhibited differential and concentration-dependent block of all of these currents. Specifically, 100 μM 4-AP selectively inhibited a slowly inactivating outward current ( K SI) that was insensitive to dendrototoxin (≤10 μM) and that activated at −50 mV. At 2 mM, 4-AP inhibited fast-inactivating, low-threshold (−70 mV) A-type currents ( K A) and sustained, TEA-sensitive noninactivating delayed-rectifier-type currents ( K DR). At an even higher concentration (8 mM), 4-AP additionally blocked inwardly rectifying, Cs+- and Ba2+-sensitive K+ currents ( K IR). Current injection into current-clamped astrocytes in culture or in acute spinal cord slices induced an overshooting voltage response reminiscent of slow neuronal action potentials. Increasing concentrations of 4-AP selectively modulated different phases in the repolarization of these glial spikes, suggesting that all four K+ currents serve different roles in stabilization and repolarization of the astrocytic membrane potential. Our data suggest that 4-AP is an useful, dose-dependent inhibitor of all four astrocytic K+ channels. We show that the slowly inactivating astrocytic K+ currents, which had not been described as separate current entities in astrocytes, contribute to the resting K+ conductance and may thus be involved in K+ homeostatic functions of astrocytes. The high sensitivity of these currents to micromolar 4-AP suggests that application of 4-AP to inhibit neuronal A-currents or to induce epileptiform discharges in brain slices also may influence astrocytic K+ buffering.
2-Deoxyglucose–Induced Long-Term Potentiation in CA1 Is Not Prevented by Intraneuronal Chelator
Authors: Zhao Yong-Tao , Krnjević Krešimir
Published: December 2017

In hippocampal slices, temporary (10–20 min) replacement of glucose with 10 mM 2-deoxyglucose is followed by marked and very sustained potentiation of EPSPs (2-DG LTP). To investigate its mechanism, we examined 2-DG's effect in CA1 neurons recorded with sharp 3 M KCl electrodes containing a strong chelator, 50 or 100 mM ethylene glycol-bis(β-aminoethyl ether)- N, N, N′, N′-tetraacetic acid (EGTA). In most cases, field EPSPs were simultaneously recorded and conventional LTP was also elicited in some cells by tetanic stimulation of stratum radiatum. 2-DG potentiated intracellular EPSP slopes by 48 ± 5.1% (SE) in nine cells recorded with plain KCl electrodes and by 52 ± 6.2% in seven cells recorded with EGTA-containing electrodes. In four of the latter cells, tetanic stimulation (twice 100 Hz for 1 s) failed to evoke LTP (2 ± 1.1%), although field EPSPs were clearly potentiated (by 28 ± 6.9%). Thus unlike tetanic LTP, 2-DG LTP is not readily prevented by postsynaptic intraneuronal injection of EGTA. These findings agree with other evidence that the rise in postsynaptic (somatic) [Ca2+]i caused by 2-DG is not the principal trigger for the subsequent 2-DG LTP and that it may be a purely presynaptic phenomenon.
Synaptic GABAA Activation Inhibits AMPA-Kainate Receptor–Mediated Bursting in the Newborn (P0–P2) Rat Hippocampus
Authors: Lamsa Karri , Palva J. Matias , Ruusuvuori Eva , Kaila Kai , Taira Tomi
Published: December 2017

The mechanisms of synaptic transmission in the rat hippocampus at birth are assumed to be fundamentally different from those found in the adult. It has been reported that in the CA3-CA1 pyramidal cells a conversion of “silent” glutamatergic synapses to conductive α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) synapses starts gradually after P2. Further, GABA via its depolarizing action seems to give rise to grossly synchronous yet slow calcium oscillations. Therefore, GABA is generally thought to have a purely excitatory rather than an inhibitory role during the first postnatal week. In the present study field potential recordings and gramicidin perforated and whole cell clamp techniques as well as K+-selective microelectrodes were used to examine the relative contributions of AMPA and GABAA receptors to network activity of CA3-CA1 pyramidal cells in the newborn rat hippocampus. As early as postnatal day( P 0–P2), highly coherent spontaneous firing of CA3 pyramidal cells was seen in vitro. Negative-going extracellular spikes confined to periodic bursts (interval 16 ± 3 s) consisting of 2.9 ± 0.1 spikes were observed in stratum pyramidale. The spikes were accompanied by AMPA-R–mediated postsynaptic currents (PSCs) in simultaneously recorded pyramidal neurons (7.6 ± 3.0 unitary currents per burst). In CA1 pyramidal cells synchronous discharging of CA3 circuitry produced a barrage of AMPA currents at >20 Hz frequencies, thus demonstrating a transfer of the fast CA3 network activity to CA1 area. Despite its depolarizing action, GABAA-R–mediated transmission appeared to exert inhibition in the CA3 pyramidal cell population. The GABAA-R antagonist bicuculline hypersynchronized the output of glutamatergic CA3 circuitry and increased the network-driven excitatory input to the pyramidal neurons, whereas the GABAA-R agonist muscimol (100 nM) did the opposite. However, the occurrence of unitary GABAA-R currents was increased after muscimol application from 0.66 ± 0.16 s−1 to 1.43 ± 0.29 s−1. It was concluded that AMPA synapses are critical in the generation of spontaneous high-frequency bursts in CA3 as well as in CA3-CA1 transmission as early as P0–P2 in rat hippocampus. Concurrently, although GABAA-R–mediated depolarization may excite hippocampal interneurons, in CA3 pyramidal neurons it can restrain excitatory inputs and limit the size of the activated neuronal population.
Summation of Effective Synaptic Currents and Firing Rate Modulation in Cat Spinal Motoneurons
Authors: Powers Randall K. , Binder Marc D.
Published: December 2017

The aim of this study was to examine how cat spinal motoneurons integrate the synaptic currents generated by the concurrent activation of large groups of presynaptic neurons. We obtained intracellular recordings from cat triceps surae motoneurons and measured the effects of repetitive activity in different sets of presynaptic neurons produced by electrical stimulation of descending fibers or peripheral nerves and by longitudinal vibration of the triceps surae muscles (to activate primary muscle spindle Ia afferent fibers). We combined synaptic activation with subthreshold injected currents to obtain estimates of effective synaptic currents at the resting potential ( I Nrest) and at the threshold for repetitive discharge ( I Nthresh). We then superimposed synaptic activation on suprathreshold injected current steps to measure the synaptically evoked change in firing rate. We studied eight different pairs of synaptic inputs. When any two synaptic inputs were activated concurrently, both the effective synaptic currents ( I Nrest) and the synaptically evoked changes in firing rate generally were equal to or slightly less than the linear sum of the effects produced by activating each input alone. However, there were several instances in which the summation was substantially less than linear. In some motoneurons, we induced a partial blockade of potassium channels by adding tetraethylammonium (TEA) or cesium to the electrolyte solution in the intracellular pipette. In these cells, persistent inward currents were evoked by depolarization that led to instances of substantially greater-than linear summation of injected and synaptic currents. Overall our results indicate that the spatial distribution of synaptic boutons on motoneurons acts to minimize electrical interactions between synaptic sites permitting near linear summation of synaptic currents. However, modulation of voltage-gated conductances on the soma and dendrites of the motoneuron can lead to marked nonlinearities in synaptic integration.
Voltage-Gated Potassium Channels Activated During Action Potentials in Layer V Neocortical Pyramidal Neurons
Authors: Kang Jian , Huguenard John R. , Prince David A.
Published: December 2017

To investigate voltage-gated potassium channels underlying action potentials (APs), we simultaneously recorded neuronal APs and single K+ channel activities, using dual patch-clamp recordings (1 whole cell and 1 cell-attached patch) in single-layer V neocortical pyramidal neurons of rat brain slices. A fast voltage-gated K+ channel with a conductance of 37 pS (Kf) opened briefly during AP repolarization. Activation of Kf channels also was triggered by patch depolarization and did not require Ca2+influx. Activation threshold was about −20 mV and inactivation was voltage dependent. Mean duration of channel activities after single APs was 6.1 ± 0.6 ms (mean ± SD) at resting membrane potential (−64 mV), 6.7 ± 0.7 ms at −54 mV, and 62 ± 15 ms at −24 mV. The activation and inactivation properties suggest that Kf channels function mainly in AP repolarization but not in regulation of firing. Kf channels were sensitive to a low concentration of tetraethylammonium (TEA, 1 mM) but not to charybdotoxin (ChTX, 100 nM). Activities of A-type channels (KA) also were observed during AP repolarization. KA channels were activated by depolarization with a threshold near −45 mV, suggesting that KA channels function in both repolarization and timing of APs. Inactivation was voltage dependent with decay time constants of 32 ± 6 ms at −64 mV (rest), 112 ± 28 ms at −54 mV, and 367 ± 34 ms at −24 mV. KA channels were localized in clusters and were characterized by steady-state inactivation, multiple subconductance states (36 and 19 pS), and inhibition by 5 mM 4-aminopyridine (4-AP) but not by 1 mM TEA. A delayed rectifier K+ channel (Kdr) with a unique conductance of 17 pS was recorded from cell-attached patches with TEA/4-AP-filled pipettes. Kdrchannels were activated by depolarization with a threshold near −25 mV and showed delayed long-lasting activation. Kdr channels were not activated by single action potentials. Large conductance Ca2+-activated K+ (BK) channels were not triggered by neuronal action potentials in normal slices and only opened as neuronal responses deteriorated (e.g., smaller or absent spikes) and in a spike-independent manner. This study provides direct evidence for different roles of various K+ channels during action potentials in layer V neocortical pyramidal neurons. Kf and KA channels contribute to AP repolarization, while KA channels also regulate repetitive firing. Kdr channels also may function in regulating repetitive firing, whereas BK channels appear to be activated only in pathological conditions.