Compound Action Potential Recovery Research Articles

Energy Metabolism in Mouse Sciatic Nerve A Fibres during Increased Energy Demand.

The ability of sciatic nerve A fibres to conduct action potentials relies on an adequate supply of energy substrate, usually glucose, to maintain necessary ion gradients. Under our ex vivo experimental conditions, the absence of exogenously applied glucose triggers Schwann cell glycogen metabolism to lactate, which is transported to axons to fuel metabolism, with loss of the compound action potential (CAP) signalling glycogen exhaustion. The CAP failure is accelerated if tissue energy demand is increased by high-frequency stimulation (HFS) or by blocking lactate uptake into axons using cinnemate (CIN). Imposing HFS caused CAP failure in nerves perfused with 10 mM glucose, but increasing glucose to 30 mM fully supported the CAP and promoted glycogen storage. A combination of glucose and lactate supported the CAP more fully than either substrate alone, indicating the nerve is capable of simultaneously metabolising each substrate. CAP loss resulting from exposure to glucose-free artificial cerebrospinal fluid (aCSF) could be fully reversed in the absence of glycogen by addition of glucose or lactate when minimally stimulated, but imposing HFS resulted in only partial CAP recovery. The delayed onset of CAP recovery coincided with the release of lactate by Schwann cells, suggesting that functional Schwann cells are a prerequisite for CAP recovery.

Microglial depletion abolishes ischemic preconditioning in white matter.

Ischemic preconditioning (IPC) is a phenomenon whereby a brief, non-injurious ischemic exposure enhances tolerance to a subsequent ischemic challenge. The mechanism of IPC has mainly been studied in rodent stroke models where gray matter (GM) constitutes about 85% of the cerebrum. In humans, white matter (WM) is 50% of cerebral volume and is a critical component of stroke damage. We developed a novel CNS WM IPC model using the mouse optic nerve (MON) and identified the involved immune signaling pathways. Here we tested the hypothesis that microglia are necessary for WM IPC. Microglia were depleted by treatment with the colony stimulating factor 1 receptor (CSF1R) inhibitor PLX5622. MONs were exposed to transient ischemia in vivo, acutely isolated 72 h later, and subjected to oxygen-glucose deprivation (OGD) to simulate a severe ischemic injury (i.e., stroke). Functional and structural axonal recovery was assessed by recording compound action potentials (CAPs) and by microscopy using quantitative stereology. Microglia depletion eliminated IPC-mediated protection. In control mice, CAP recovery was improved in preconditioned MONs compared with non-preconditioned MONs, however, in PLX5622-treated mice, we observed no difference in CAP recovery between preconditioned and non-preconditioned MONs. Microgliadepletion also abolished IPC protective effects on axonal integrity and survival of mature (APC+ ) oligodendrocytes after OGD. IPC-mediated protection was independent of retinal injury suggesting it results from mechanistic processes intrinsic to ischemia-exposed WM. We conclude that preconditioned microglia are critical for IPC in WM. The "preconditioned microglia" phenotype might protect against other CNS pathologies and is a neurotherapeutic horizon worth exploring.

Hypothermic neuroprotection during reperfusion following exposure to aglycemia in central white matter is mediated by acidification.

Hypoglycemia is a common iatrogenic consequence of type 1 diabetes therapy that can lead to central nervous system injury and even death if untreated. In the absence of clinically effective neuroprotective drugs we sought to quantify the putative neuroprotective effects of imposing hypothermia during the reperfusion phase following aglycemic exposure to central white matter. Mouse optic nerves (MONs), central white matter tracts, were superfused with oxygenated artificial cerebrospinal fluid (aCSF) containing 10 mmol/L glucose at 37°C. The supramaximal compound action potential (CAP) was evoked and axon conduction was assessed as the CAP area. Extracellular lactate was measured using an enzyme biosensor. Exposure to aglycemia, simulated by omitting glucose from the aCSF, resulted in axon injury, quantified by electrophysiological recordings, electron microscopic analysis confirming axon damage, the extent of which was determined by the duration of aglycemia exposure. Hypothermia attenuated injury. Exposing MONs to hypothermia during reperfusion resulted in improved CAP recovery compared with control recovery measured at 37°C, an effect attenuated in alkaline aCSF. Hypothermia decreases pH implying that the hypothermic neuroprotection derives from interstitial acidification. These results have important clinical implications demonstrating that hypothermic intervention during reperfusion can improve recovery in central white matter following aglycemia.

Effect of DSPE-PEG on compound action potential, injury potential and ion concentration following compression in ex vivo spinal cord

It has been shown that polyethylene glycol (PEG) can reseal membrane disruption on the spinal cord, but only high concentrations of PEG have been shown to have this effect. Therefore, the effect of PEG is somewhat limited, and it is necessary to investigate a new approach to repair spinal cord injury. This study assesses the ability of 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(poly (ethylene glycol)) 2000] (DSPE-PEG) to recover physiological function and attenuate the injury-induced influx of extracellular ions in ex vivo spinal cord injury. Isolated spinal cords were subjected to compression injury and treated with PEG or DSPE-PEG immediately after injury. The compound action potential (CAP) was recorded before and after injury to assess the functional recovery. Furthermore, injury potential, the difference in gap potentials before and after compression, and the concentration of intracellular ions were used to evaluate the effect of DSPE-PEG on reducing ion influx. Data showed that the injury potential and ion concentration of the untreated, PEG and DSPE-PEG group, without significant difference among them, are remarkably higher than those of the intact group. Moreover, the CAP recovery of the DSPE-PEG and PEG treated spinal cords was significantly greater than that of the untreated spinal cords. The level of CAP recovery in the DSPE-PEG and PEG treated groups was the same, but the concentration of DSPE-PEG used was much lower than the concentration of PEG. These results suggest that instant application of DSPE-PEG could effectively repair functional disturbance in SCI at a much lower concentration than PEG.

The Neuroprotective Ability of Polyethylene Glycol is Affected by Temperature in Ex Vivo Spinal Cord Injury Model

Immediate membrane sealing after spinal cord injury (SCI) can prevent further degradation and result in ultimate functional recovery. It has been reported that polyethylene glycol (PEG) can repair membrane damage caused by mechanical insults to the spinal cord. Furthermore, membrane fluidity and its sealing process vary at different temperatures. Here, we have assessed the possible synergistic effects of PEG and temperature on the repair of neural membranes in an SCI model. The effects of PEGs (400, 1,000 and 2,000 Da) were studied at different temperatures (25, 37 and 40 °C) by means of compound action potential (CAP) recovery and a lactate dehydrogenase (LDH) assay. Isolated spinal cords were mounted in a double sucrose gap chamber, where the amplitude and area of CAPs were recorded after implementing injury, in the presence and absence of PEG. Moreover, the LDH assay was used to assess the effects of PEG on membrane resealing. Data showed that the least CAP recovery occurred at 25 °C, followed by 37 and 40 °C, in all treated groups. Moreover, maximum CAP amplitude recovery, 65.46 ± 5.04 %, was monitored in the presence of PEG400 at 40 °C, followed by 41.49 ± 2.41 % in PEG1000 and 37.36 ± 1.62 % in PEG2000. Furthermore, raising the temperature from 37 to 40 °C significantly increased CAP recovery in the PEG2000 group. Similar recovery patterns were obtained by CAP area measurements and LDH assay. The results suggest that application of low-molecular weight PEG (PEG400) in mild hyperthermia conditions (40 °C) provides the optimum condition for membrane sealing in SCI model.

Effects of the Noradrenergic System in Rat White Matter Exposed to Oxygen–Glucose DeprivationIn Vitro

Norepinephrine (NE) is released in excess into the extracellular space during oxygen-glucose deprivation (OGD) in brain, increasing neuronal metabolism and aggravating glutamate excitoxicity. We used isolated rat optic nerve and spinal cord dorsal columns to determine whether the noradrenergic system influences axonal damage in white matter. Tissue was studied electrophysiologically by recording the compound action potential (CAP) before and after exposure to 60 min of OGD at 36 degrees C. Depleting catecholamine stores with reserpine was protective and improved CAP recovery after 1 h of reperfusion from 17% (control) to 35%. Adding NE during OGD decreased CAP recovery to 8%, and adding NE to reserpine during OGD eliminated the protective effect of the latter. Selective inhibitors of Na(+)-dependent norepinephrine transport desipramine and nisoxetine improved recovery to 58% and 44%, respectively. alpha2 adrenergic receptor agonists UK14,304 and medetomidine improved CAP recovery to 41% and 46% after 1 h of OGD. Curiously, alpha2 antagonists alone were also highly protective (e.g., atipamezole: 86% CAP recovery), at concentrations that did not affect baseline excitability. The protective effect of alpha2 receptor modulation was corroborated by imaging fluorescent Ca(2+) and Na(+) indicators within axons during OGD. Both agonists and antagonists significantly reduced axonal Ca(2+) and Na(+) accumulation in injured axons. These data suggest that the noradrenergic system plays an active role in the pathophysiology of axonal ischemia and that alpha2 receptor modulation may be useful against white matter injury.

Critical roles of decompression in functional recovery of ex vivo spinal cord white matter

The correlations between functional deficits, the magnitude of compression, and the role of sustained compression during traumatic spinal cord injury remain largely unknown. Thus, the functional outcome of this type of injury with or without surgical intervention is rather unpredictable. To elucidate how severity and duration of compression affect cord function, the authors have developed a method to study electrophysiological characteristics and axonal membrane damage in white matter from guinea pig spinal cord. Ventral white matter strips isolated from adult guinea pigs were compressed by a compression rod at a level of either 60 or 80% and held briefly, for 30 minutes, or for 60 minutes. In half the experimental groups, a decompression phase consisting of probe withdrawal and 30 minutes of recovery was also applied. For all cord samples, functional response was continuously monitored through compound action potential (CAP) recording. In addition, axonal membrane damage was assessed by a horseradish peroxidase (HRP) exclusion assay. After 30 minutes of sustained compression at levels of 60 or 80%, a spinal cord decompression procedure caused a significant CAP recovery, with specimens reaching 97.5 +/- 6.84% (p < 0.05) and 56.2 +/- 6.14% (p < 0.05) of preinjury amplitude, respectively. After 60 minutes of compression, the amount of CAP recovery following the decompression stage was only 65.5 +/- 9.33% for 60% compression (p < 0.05) and 29.8 +/- 6.31% for 80% compression (p < 0.05). Unlike the CAP response, HRP uptake did not increase during sustained compression, and the data showed that HRP staining was primarily time dependent. The degree of axonal membrane damage is not exacerbated during sustained compression. However, the electrical conductivity of the cord white matter weakens throughout the duration of compression. Therefore, decompression is a viable procedure for preservation of neurological function following compressive injury.

Physiological effects of auditory nerve myelinopathy in chinchillas

The goals were to study the physiological effects of auditory nerve myelinopathy in chinchillas and to test the hypothesis that myelin abnormalities could account for auditory neuropathy, a hearing disorder characterized by absent auditory brainstem responses (ABRs) with preserved outer hair cell function. Doxorubicin, a cytotoxic drug used as an experimental demyelinating agent, was injected into the auditory nerve bundle of 18 chinchillas; six other chinchillas were injected with vehicle alone. Cochlear microphonics, compound action potentials (CAPs), inferior colliculus evoked potentials (IC-EVPs), cubic distortion product otoacoustic emissions and ABRs were recorded before and up to 2 months after injection. Cochleograms showed no hair cell loss in any of the animals and measures of outer hair cell function were normal (cubic distortion product otoacoustic emissions) or enhanced (cochlear microphonics) after injection. ABR was present in animals with mild myelin damage (n = 10) and absent in animals with severe myelin damage that included the myelin surrounding spiral ganglion cell bodies and fibers in Rosenthal's canal (n = 8). Animals with mild damage had reduced response amplitudes at 1 day, followed by recovery of CAP and enhancement of the IC-EVP. In animals with severe damage, CAP and IC-EVP thresholds were elevated, amplitudes were reduced, and latencies were prolonged at 1 day and thereafter. CAPs deteriorated over time, whereas IC-EVPs partially recovered; latencies remained consistently prolonged despite changes in amplitudes. The results support auditory nerve myelinopathy as a possible pathomechanism of auditory neuropathy but indicate that myelinopathy must be severe before physiological measures are affected.

Complex interplay between glutamate receptors and intracellular Ca2+ stores during ischaemia in rat spinal cord white matter

Electrophysiological recordings of propagated compound action potentials (CAPs) and axonal Ca(2+) measurements using confocal microscopy were used to study the interplay between AMPA receptors and intracellullar Ca(2+) stores in rat spinal dorsal columns subjected to in vitro combined oxygen and glucose deprivation (OGD). Removal of Ca(2+) or Na(+) from the perfusate was protective after 30 but not 60 min of OGD. TTX was ineffective with either exposure, consistent with its modest effect on ischaemic depolarization. In contrast, AMPA antagonists were very protective, even after 60 min of OGD where 0Ca(2+) + EGTA perfusate was ineffective. Similarly, blocking ryanodine receptor-mediated Ca(2+) mobilization from internal stores (0Ca(2+) + nimodipine or 0Ca(2+) + ryanodine), or inositol 1,4,5-trisphosphate (IP(3))-dependent Ca(2+) release (block of group 1 metabotropic glutamate receptors with 1-aminoindan-1,5-dicarboxylic acid, inhibition of phospholipase C with U73122 or IP(3) receptor block with 2APB; each in 0Ca(2+)) were each very protective, with the combination resulting in virtually complete functional recovery after 1 h OGD (97 +/- 32% CAP recovery versus 4 +/- 6% in artificial cerebrospinal fluid). AMPA induced a rise in Ca(2+) concentration in normoxic axons, which was greatly reduced by blocking ryanodine receptors. Our data therefore suggest a novel and surprisingly complex interplay between AMPA receptors and Ca(2+) mobilization from intracellular Ca(2+) stores. We propose that AMPA receptors may not only allow Ca(2+) influx from the extracellular space, but may also significantly influence Ca(2+) release from intra-axonal Ca(2+) stores. In dorsal column axons, AMPA receptor-dependent mechanisms appear to exert a greater influence than voltage-gated Na(+) channels on functional outcome following OGD.

Increased excitability of acidified skeletal muscle: role of chloride conductance.

Generation of the action potentials (AP) necessary to activate skeletal muscle fibers requires that inward membrane currents exceed outward currents and thereby depolarize the fibers to the voltage threshold for AP generation. Excitability therefore depends on both excitatory Na+ currents and inhibitory K+ and Cl− currents. During intensive exercise, active muscle loses K+ and extracellular K+ ([K+]o) increases. Since high [K+]o leads to depolarization and ensuing inactivation of voltage-gated Na+ channels and loss of excitability in isolated muscles, exercise-induced loss of K+ is likely to reduce muscle excitability and thereby contribute to muscle fatigue in vivo. Intensive exercise, however, also leads to muscle acidification, which recently was shown to recover excitability in isolated K+-depressed muscles of the rat. Here we show that in rat soleus muscles at 11 mM K+, the almost complete recovery of compound action potentials and force with muscle acidification (CO2 changed from 5 to 24%) was associated with reduced chloride conductance (1731 ± 151 to 938 ± 64 μS/cm2, P < 0.01) but not with changes in potassium conductance (405 ± 20 to 455 ± 30 μS/cm2, P < 0.16). Furthermore, acidification reduced the rheobase current by 26% at 4 mM K+ and increased the number of excitable fibers at elevated [K+]o. At 11 mM K+ and normal pH, a recovery of excitability and force similar to the observations with muscle acidification could be induced by reducing extracellular Cl− or by blocking the major muscle Cl− channel, ClC-1, with 30 μM 9-AC. It is concluded that recovery of excitability in K+-depressed muscles induced by muscle acidification is related to reduction in the inhibitory Cl− currents, possibly through inhibition of ClC-1 channels, and acidosis thereby reduces the Na+ current needed to generate and propagate an AP. Thus short term regulation of Cl− channels is important for maintenance of excitability in working muscle.

Recovery of human compound action potential using a paired-click stimulation paradigm

The recovery process from adaptation of the compound action potential (CAP) was studied using an equilevel paired click stimulation paradigm in humans with normal hearing. The CAP amplitude to the second click of paired clicks was normalized to the amplitude to the first click. The second-click CAP amplitude recovered as a function of interclick interval (ICI) between the first and the second click of a pair. A regression line fitted to the recovered amplitude data demonstrated the logarithmic function of the ICI. Full recovery times changed from 118 to 278 ms with increasing click intensity. The regression lines for higher click intensities exhibited two different slopes in two ICI ranges: from 3 to 100 and 120 to 300 ms. We suppose that the CAP recovery for ICI <100 ms is attributable to both the relative refractoriness of auditory nerve and the short-term adaptation mechanisms, while, for ICI >100 ms chiefly to the short-term adaptation mechanisms. The recovery process of the second-click CAP slowed with increasing intensity, which is a similar result to that obtained in the animal experiments by Parham et al. The input–output (I–O) curve of the second-click CAP amplitudes exhibited a different slopes above and below 60 dB normal hearing level (nHL). We assume that the mechanisms underlying this characteristic curve pattern differ from those for the I–O curve of the CAP in response to single-click stimuli. We expect that investigating the CAP recovery in pathological ears will provide clinically useful information on cochlear synaptic function.

Contribution to Ischemic Injury of Rat Optic Nerves by Intracellular Sodium Overload

Ischemic insult to axons of retinal ganglion cells (RGCs) is believed to contribute significantly to preferential loss of RGCs in glaucoma. In this study, we characterized the role of intracellular Na(+) overload in ischemic injury of acutely isolated rat optic nerves by evaluating electrically elicited compound action potentials (CAPs) from the optic nerves. Under control conditions, robust and stable CAPs can be recorded for more than 5 h. One hour of oxygen and glucose deprivation (OGD) that simulates ischemia, virtually eliminated the CAP. Upon returning to control conditions, the CAP gradually recovered. Maximum recovery (35% of control) was obtained by 1 h after returning to normal oxygenated Ringer. When a rapidly reversible Na(+) channel blocker, that completely blocked the CAP under control conditions, was present during OGD, the recovery of the CAP was significantly enhanced to 65% of control. When the Na(+) was replaced with either choline or Li(+) in the Ringer during OGD, CAP recovery was significantly enhanced (65-70% of control). Removing Ca(++) from the Ringer (plus 5 mM EGTA) provided even better preservation of the CAP following OGD (90% of control). Our results are consistent with the hypothesis that intracellular Na(+) overload appears to play a significant role in ischemic injury of optic nerves. This Na(+) overload may depend at least partially upon Ca(++) influx from the extracellular space.

Questioning the Mechanism of Nerve Injury

We thank Lambert et al. for their interest in our article1and stimulating comments. We would like to take the opportunity to address the issues raised by their insights.Lambert et al. suggest that the mechanism of local anesthetic neurotoxicity is unrelated to membrane disruption caused by the detergent nature of highly concentrated local anesthetics, instead resulting from breakdown of the sodium conductance system (i.e. , absence of compound action potential generation) despite the presence of an intact membrane structure and normal ionic gradients and compound resting potential. The basis for their contention is that, although both compound action potential and compound resting potential in excised sciatic nerves of the bullfrog disappear permanently after bathing in sodium lauryl sulfate (typical detergent) solution,2compound resting potential recovers without compound action potential recovery after bathing in 5% lidocaine solution for 15 min, as noted in their earlier electrophysiologic study.3However, the study cited as grounds for their question about our conclusions seems to have an inherent limitation in that the period for bathing the specimen in 5% lidocaine is 15 min. Kanai et al. 4reported a similar study using a single crayfish axon and demonstrated that although resting potential recovers after bathing in 80 mm lidocaine solution for 15 min, resting potential permanently disappears after bathing for 30 min. They suggested that highly concentrated lidocaine causes membrane disruption and indicated in that time-dependent study that sufficient exposure (≥ 30 min) was needed for membrane disruption to be induced by lidocaine. Ready et al. 5also demonstrated successfully that lidocaine at a concentration of 4% or higher induces histopathologic changes in spinal nerves during a dose-dependent study of a spinal anesthesia model in rabbits. The fact that highly concentrated lidocaine causes membrane disruption is thus not in doubt.We advocated in our article that the mechanism of membrane disruption induced by highly concentrated local anesthetics would result from the detergent nature of these agents.1From the perspective of results from various investigators, including Lambert et al. , we speculate that an intermediate step may exist in the membrane disruption dynamics induced by 5% lidocaine, with resting membrane potential maintained despite permanent inhibition of action potential. The electrophysiologic phenomena described by Lambert et al. may represent an early phase in the sequence of neurotoxic dynamics induced by 5% lidocaine.* Saga Medical School, Nabeshima, Saga, Japan. kitagawa@mail.anes.saga-med.ac.jp

Metabolic Effects of Hypothermia and Its Neuroprotective Effects on the Recovery of Metabolic and Electrophysiological Function in the Ischemic Retina in Vitro

Reduction in energy usage has been investigated as the mechanism by which hypothermia provides protection during ischemia. We describe experiments using hypothermia in the rabbit retina in vitro that show a correlation between hypothermia-induced reductions in energy usage and neuroprotection. We examined energy metabolism and electrophysiological function under control/nonischemic conditions during 1 or 2 hours of "ischemia" (induced by decreasing glucose from 6 to 1 mmol/L and oxygen from 95 to 15%) and during 3 to 4 hours of "return-to-control" conditions. Glucose utilization and lactate production were measured as indices of energy metabolism, and light-evoked compound action potentials were monitored to assess functional recovery. Nonischemic retinas subjected to both mild (33 +/- 0.5 degrees C) and moderate (30 +/- 0.5 degrees C) hypothermia exhibited a decrease of 38% in the rate of glucose utilization and lactate production compared with normothermic retinas (36 +/- 0.5 degrees C) (analysis of variance, P < 0.001). In retinas that were made ischemic, mild or moderate hypothermia further reduced the rates of glucose utilization (18 and 39%, respectively) and lactate production (21 and 28%, respectively) (P < 0.001 for glucose, P < 0.01 for lactate). Retinas that had been mildly or moderately hypothermic during ischemia exhibited improved recovery of glucose utilization (65 and 57%, respectively) and lactate production (72 and 74%, respectively) compared with normothermic retinas (18% for glucose and 54% for lactate; repeated-measures analysis of variance, P < 0.001). Recovery of compound action potentials for retinas kept at 36, 33, and 30 degrees C was 15, 36, and 53%, respectively (repeated-measures analysis of variance, P < 0.001). Our studies in an avascular neuronal model of ischemia demonstrate that hypothermia protects against ischemic injury. We interpret the smaller reductions in energy generation and usage caused by ischemia when the retinas were hypothermic as evidence that hypothermia had reduced energy requirements more than energy production, and we propose that this at least in part explains its protection.

Effects of serotonin 1A agonist on acute spinal cord injury.

We evaluated the effects of serotonin (5-HT) agonists on in vitro models of spinal cord compressive injury. Evoked potentials in injured rat spinal cords (n=24) were recorded during perfusion with 5-HT agonists. To evaluate the therapeutic effects of 5-HT agonists on the recovery of compound action potentials in injured spinal cords. Rat dorsal columns were isolated, placed in a chamber, and injured by extradural compression with a clip. Conducting action potentials were activated by supramaximal constant current electrical stimuli and recorded during perfusion with 5-HT agonists and antagonists. After inducing compression injuries, mean action potential amplitudes were reduced to 33.9+/-5.4% of the pre-injury level. After 120 min of perfusion with Ringer's solution, the mean amplitudes recovered to 62.8+/-8.4% of the pre-injury level. At a concentration of 100 micro M, perfusion with tandospirone (a 5-HT1A agonist) resulted in a significantly greater recovery of mean action potential amplitudes at 2 h after the injury (86.2+/-6.9% of pre-injury value) as compared with the control Ringer's solution (62.8+/-8.4% of pre-injury value, P<0.05). In contrast, quipazine (a 5-HT2A agonist) accelerated the decrease of amplitude (54.5+/-11.7% of pre-injury value). 5-HT1A and 5-HT2A agonist did not consistently alter latencies of the action potentials. The 5-HT1A receptor agonist was effective for the recovery of spinal action potential amplitudes in a rat spinal cord injury model.

Endocochlear potentials and compound action potential recovery: functions in the C57BL/6J mouse

The C57BL/6J mouse suffers from cochlear degeneration beginning at an early age and has been used as a model of age-related hearing loss (presbyacusis). Here, the endocochlear potential (EP) and compound action potential (CAP) responses were determined in one-, four-, 12- and 24-month-old C57BL/6J mice. CAP measures included thresholds to tone pips, input/output (I/O) functions, and recovery functions to conditioning tones. EP values among the four age groups did not differ significantly ( P>0.05) in either the basal or apical turns. CAP thresholds were increased significantly by 10 to 30 dB in the four-month group compared to the one-month controls at 11.3, 16, 20, and 22.6 kHz. CAP I/O functions were shallower in the four-month group compared to controls at all frequencies. In the 12- and 24-month-old mice, CAP responses were absent, despite normal EP values in these animals. Recovery functions after conditioning tones were obtained at 8, 16, 20 and 22.6 kHz; the functions had fast and slow components at all frequencies tested in both the one- and four-month-old groups. The corresponding recovery curves were identical for both age groups, even with significant threshold shifts in the older group. The two component recovery curves provide the first physiological evidence that different spontaneous rate (SR) classes of auditory neurons exist in the C57BL/6J mouse. Moreover, the unchanged recovery functions in the older group suggest that there was no loss of activity of the low-SR fiber population with age under conditions where the EP remains stable, in contrast to the gerbil model of presbyacusis where there is a loss of low-SR fiber activity and EP does decline with age.

Ionic mechanisms of aglycemic axon injury in mammalian central white matter.

The authors investigated ionic mechanisms underlying aglycemic axon injury in adult rat optic nerve, a central white matter tract. Axon function was assessed using evoked compound action potentials (CAPs). Glucose withdrawal led to delayed CAP failure, an alkaline extracellular pH shift, and an increase in extracellular [K(+)]. Sixty minutes of glucose withdrawal led to irreversible axon injury. Aglycemic axon injury required extracellular calcium; the extent of injury progressively declined as bath [Ca(2+)] was decreased. To evaluate Ca(2+) movements during aglycemia, the authors recorded extracellular [Ca(2+)] ([Ca(2+)](o)) using Ca(2+)-sensitive microelectrodes. Under control conditions, [Ca(2+)](o) fell with a similar time course to CAP failure, indicating extracellular Ca(2+) moved to an intracellular position during aglycemia. The authors quantified the magnitude of [Ca(2+)]o decrease as the area below baseline [Ca(2+)]o during aglycemia and used this as a qualitative measure of Ca(2+) influx. The authors studied the mechanisms of Ca(2+) influx. Blockade of Na(+) influx reduced Ca(2+) influx and improved CAP recovery, suggesting Na(+)-Ca(2+) exchanger involvement. Consistent with this hypothesis, bepridil reduced axon injury. In addition, diltiazem or nifedipine decreased Ca(2+) influx and increased CAP recovery. The authors conclude aglycemic central white matter injury is caused by Ca(2+) influx into intracellular compartments through reverse Na(+)-Ca(2+) exchange and L-type Ca(2+) channels.

Electrocochleogram after transection of vestibulo-cochlear nerve in a patient with a large acoustic neurinoma

This study reports pre- and post-operative compound action potentials (CAPs) that were recorded from a 27-year-old woman with an acoustic neurinoma. During surgery it was necessary to totally sever her vestibulo-cochlear nerve to excise a large tumor. A pure tone audiogram changed to the scale-out pattern immediately after operation. However, CAP, the waveform of which was broadened, has been recorded 3 years post-operatively with a threshold elevation of 10 dB over the pre-operative threshold. This phenomenon suggests that CAP may originate from the extreme periphery of the auditory nerve within the cochlea. The broadening of the CAP was assumed to result from enhancement of the negative summating potential included in the CAP. We studied the effect of preceding stimulus on CAP using paired click stimuli pre- and post-operatively. A reduction of CAP amplitude in response to the second click of paired clicks was markedly suppressed in the inter-click interval between 3 and 80 ms post-operatively. We speculate that depletion of adaptation induced the abnormal CAP recovery described above and that the lateral efferent nerve system was involved in abnormal CAP adaptation with transection of the vestibulo-cochlear nerve in this case.

Disruption of cochlear potentials by chemical asphyxiants: Cyanide and carbon monoxide

While ischemia, hypoxic hypoxia, and carbon monoxide (CO) have received extensive study designed to characterize mechanisms by which they disrupt cochlear function, little data are available concerning cyanide's potential to disrupt auditory function. In this study, disruption of the compound action potential (CAP) and endocochlear potential (EP) by cyanide and CO was compared in rats treated with potassium cyanide (KCN) (7 mg/kg ip), saline, CO (35 ml/kg ip), and air. Acute KCN administration significantly suppressed CAP and EP transiently. The effect was seen initially on EP with CAP impairment occurring a few minutes later. Acute CO injection also suppressed the CAP significantly, but the effect was far smaller, occurred later in time, and lasted longer than the effect of KCN. The effect of CO on EP was equivocal. There was a good correspondence between blood cyanide levels and impairment of cochlear function; carboxyhemoglobin (HbCO) levels were elevated during the period when cochlear function was impaired, but recovery of cochlear function preceded the return of normal oxyhemoglobin. Both KCN and CO had somewhat preferential effects on high-frequency tones. Repeated cyanide administration caused a persistent CAP threshold elevation despite the rapid recovery of EP and CAP observed following acute KCN administration. The data suggest that acute KCN administration has a prominent disruptive effect at the stria vascularis presumably by disrupting the electron transport chain in this metabolically active structure. The principal target for acute CO ototoxicity in the cochlea is probably not the stria vascularis.

Recovery from adaptation in human compound action potential following forward masking

Recovery from adaptation was studied as changes in compound action potential (CAP) in response to clicks after broadband noise masking in volunteers with normal hearing. CAP was transtympanically elicited from the promontory using a needle electrode. Preceding masking noise and subsequent click stimuli were delivered separately from 2 loud speakers 80 cm from the tested ear. We evaluated the effect of masking noise duration and intensity on CAP recovery from adaptation as a function of delta t, defined as the interval between masking noise offset and click stimulus onset. Results were as follows: At delta t shorter than 200 ms or less, recovery from latency prolongation and amplitude decrease slowed with increasing masking noise duration and intensity. At delta t longer than 200 ms, no significant difference was seen in CAP recovery based on changes in masking noise duration or intensity. Recovery from adaptation, which depended on click intensity, took about 700 ms at a 40 dBnHL click and about 300 ms at a 60 dBnHL click. The influence of the middle ear muscle reflex on CAP recovery patterns is neglected when considering the intensity both masking noise and click stimulus. In CAP recovery, accumulated effects of the relative auditory-nerve refractory period may adversely affect recovery at very short delta t, while recovery from short-term adaptation, assumed involved in cochlear-hair cell synapse function, may be an important factor at delta t longer than 30-40 ms. Based on animal experiments showing different recovery times dependent on different auditory nerve spontaneous rates(SR), we concluded that at delta t of 200 ms or less, CAP recovery reflects high SR neuron activity, while at delta t longer than 200 ms, it mainly reflects that of low SR neurons.