Increasing Stimulus Frequency Research Articles

Paced breathing and respiratory movement responses evoked by bidirectional constant current stimulation in anesthetized rabbits.

Objective: Diaphragm pacing (DP) is a long-term and effective respiratory assist therapy for patients with central alveolar hypoventilation and high cervical spinal cord injury. The existing DP system has some limitations, especially high price, inconvenience preoperative evaluation methods and diaphragm fatigue easily. Our objective was to develop a DP system and evaluated reliability through hardware testing and animal experiments. Methods: A DP system with bidirectional constant current was designed, manufactured and tested. Effects of a wide range of stimulus amplitudes (range: .5-2.5mA) and frequencies (range: 10-250Hz) on airflow and corresponding inspired volume were investigated during DP. Differences in airflow characteristics under various stimulation parameters were evaluated using power function. ECG interference in diaphragm electromyography (EMGdi) was filtered out using stationary wavelet transform to obtain pure EMGdi (EMGdip). 80-min period with a tendency for diaphragm fatigue by root mean square (RMS) and centroid frequency (f c ) of EMGdip was studied. Results: The increase of stimulus frequency and amplitude in animals resulted in different degrees of increase in envoked volume. Significant difference in Airflow Index (b) between anesthesia and DP provided a simple, non-invasive and feasible solution for phrenic nerve conduction function test. Increased stimulation duration with the developed DP system caused less diaphragm fatigue. Conclusion: A modular, inexpensive and reliable DP was successfully developed. Its effectiveness was confirmed in animal experiments. Significance: This study is useful for design of future implantable diaphragmatic pacemakers for improving diaphragm fatigue and convenient assessment of respiratory activity in experiments.

Role of adenosine triphosphate and protein kinase A in the force-frequency relationship in isolated rat cardiomyocytes

The physiological heart rate of rodents is around 4-6 Hz, although a stimulus frequency of 1 Hz is generally used in isolated cardiomyocytes to study changes in the contraction-relaxation cycle in cardiac muscle physiology and pathophysiology. Our study investigated the contraction parameters in isolated cardiomyocytes at 1, 2 and 4 Hz stimulation, and the roles of ATP and protein kinase A (PKA) in the force-frequency relationship in isolated cardiomyocytes. The contraction of the cell and intracellular Ca2+ changes were recorded simultaneously during cell stimulation by applying pulses of 6-8 V amplitude with frequencies of 1, 2 and 4 Hz. The increase in stimulus frequency caused a significant decrease in the percentage of shortening, relaxation times, slowing of the relaxation rate, and a significant increase in diastolic Ca2+ levels, but had no effect on the contraction rate and Ca2+ transients. Administration of ATP and N6-benzoyladenosine-3'-5'-cyclic monophosphate (6-BNZ-cAMP) caused an increase in contraction amplitude and speed which were proportional to the stimulus frequency but had no effect on the relaxation times. The experimental results show that the force-stimulus frequency has a negative correlation in isolated myocytes and that energy metabolism and the ?-adrenergic system may be responsible for this relationship.

Retinal ganglion cell desensitization is mitigated by varying parameter constant excitation pulse trains.

Retinal prostheses partially restore vision in patients blinded by retinitis pigmentosa (RP) and age-related macular degeneration (AMD). One issue that limits the effectiveness of retinal stimulation is the desensitization of the retina response to repeated pulses. Rapid fading of percepts is reported in clinical studies. We studied the retinal output evoked by fixed pulse trains vs. pulse trains that have variable parameters pulse-to-pulse. We used the current clamp to record RGC spiking in the isolated mouse retina. Trains of biphasic current pulses at different frequencies and amplitudes were applied. The main results we report are: (1) RGC desensitization was induced by increasing stimulus frequency, but was unrelated to stimulus amplitude. Desensitization persisted when the 20 Hz stimulation pulses were applied to the retinal ganglion cells at 65 μA, 85 μA, and 105 μA. Subsequent pulses in the train evoked fewer spikes. There was no obvious desensitization when 2 Hz stimulation pulse trains were applied. (2) Blocking inhibitory GABAA receptor increased spontaneous activity but did not reduce desensitization. (3) Pulse trains with constant charge or excitation (based on strength-duration curves) but varying pulse width, amplitude, and shape increased the number of evoked spikes/pulse throughout the pulse train. This suggests that retinal desensitization can be partially overcome by introducing variability into each pulse.

Intraoperative Electrocochleography in Subjects Affected by Vestibular Schwannoma and Ménière’s Disease: Comparison of Results

Objectives: Histologic reports of temporal bones of ears with vestibular schwannomas (VSs) have indicated findings of endolymphatic hydrops (ELH) in some cases. The main goal of this investigation was to test ears with VSs to determine if they exhibit electrophysiological characteristics similar to those of ears expected to experience ELH. Design: Fifty-three subjects with surgically confirmed VS aged ≥18 and with normal middle ear status were included in this study. In addition, a second group of adult subjects (n = 24) undergoing labyrinthectomy (n = 6) or endolymphatic sac decompression and shunt (ELS) placement (n = 18) for poorly controlled vestibular symptoms associated with Meniere’s disease (MD) participated in this research. Intraoperative electrocochleography (ECochG) from the round window was performed using tone burst stimuli. Audiometric testing and word recognition scores (WRS) were performed preoperatively. ECochG amplitudes, cochlear microphonic/auditory nerve neurophonic (ANN) in the form of the “ongoing” response and summation potential, were analyzed and compared between the two groups of subjects. In addition, to evaluate any effect of auditory nerve function, the auditory nerve score was calculated for each subject. Pure-tone averages were obtained using the average air conduction thresholds at 0.5, 1, and 2 kHz while WRS was assessed using Northwestern University Auditory Test No. 6 word lists. Results: In the VS group the average pure-tone averages and WRS were 59.6 dB HL and 44.8%, respectively, while in the MD group they were 52.3 dB HL and 73.8%. ECochG findings in both groups revealed a reduced trend in amplitude of the ongoing response with increased stimulus frequency. The summation potential amplitudes of subjects with VS were found to be less negative than the MD subjects for nearly all test frequencies. Finally, the VS group exhibited poorer amounts of auditory nerve function compared to the MD group. Conclusions: The current findings suggest cochlear pathology (e.g., hair cell loss) in both groups but do not support the hypothesis that VSs cause ELH.

Impaired myosin isoform shift and calcium transients contribute to cellular pathogenesis of rat cirrhotic cardiomyopathy.

Cirrhotic cardiomyopathy is a recently recognized entity, but detailed cellular and molecular mechanisms remain unclarified. We aimed to elucidate the role of myosin heavy chain isoform shifts and their relation to calcium transients in the contractile kinetics of cirrhotic rats. Cirrhosis was induced in male Lewis Brown-Norway rats by bile duct ligation (BDL). Myosin heavy chain (MHC) isoform distribution was evaluated by gel electrophoresis. Contractile force, Ca2+ transients and cell shortening were studied at varied frequency and extracellular [Ca2+ ]. T-tubular integrity was analysed by power spectrum analysis of images of myocytes stained with di-8-ANEPPS. Compared with sham controls, the phenotypes of cirrhotic rats were as follows: (a) alpha-myosin heavy chain shifted to beta-MHC isoform; (b) mild loss of T-tubular integrity in myocytes; (c) a reduced maximum and rate of rise of the Ca2+ transient (max F/Fo ); (d) a reduction in both the rate of rise and fall of contraction; (e) decreased maximal force-generating capacity; (f) loss of the inotropic effect of increased stimulus frequency; (g) unchanged sensitivity of force development to varied extracellular [Ca2+ ] and (h) increased spontaneous diastolic sarcomere length fluctuations. Cardiomyocytes and ventricular trabeculae in a cirrhotic rat model showed features of typical heart failure including systolic and diastolic prolongation, impaired force-frequency relation and decreased force-generating capacity. Impaired myosin isoform shift and calcium transients are important contributory mechanisms underlying the pathogenesis of the heart failure phenotype seen in cirrhosis.

Frequency-dependent response in cortical network with periodic electrical stimulation.

Electrical stimulation can shape oscillations in brain activity. However, the mechanism of how periodic electrical stimulation modulates brain oscillations by time-delayed neural networks is poorly understood at present. To address this question, we investigate the effects of periodic stimulations on the oscillations generated via a time-delayed neural network. We specifically study the effect of unipolar and asymmetric bidirectional pulse stimulations by altering amplitude and frequency in a systematic manner. Our findings suggest that electrical stimulations play a central role in altering oscillations in the time-delayed neural network and that these alterations are strongly dependent on the stimulus frequency. We observe that the time-delayed neural network responds differently as the stimulation frequency is altered, as manifested by changes in resonance, entrainment, non-linear oscillation, or oscillation suppression. The results also indicate that the network presents similar response activities with increasing stimulus frequency under different excitation-inhibition ratios. Collectively, our findings pave the way for exploring the potential mechanism underlying the frequency-dependent modulation of network activity via electrical stimulations and provide new insights into possible electrical stimulation therapies to the neurological and psychological disorders in clinical practice.

Response Profiles of Retinal Ganglion Cells to Sinusoidal Electric Stimulation vary for Low vs. High Frequencies.

Microelectronic retinal prostheses electrically stimulate retinal neurons with the goal of restoring vision in patients blinded by outer retinal degeneration. Despite some success in clinical trials, the quality of vision elicited by these devices is still limited. To improve the performance of retinal prostheses, our group studied how retinal neurons respond to electric stimulation. Our previous work showed that responses of retinal ganglion cells (RGCs) are frequency-dependent and different types of RGCs can be preferentially activated with a specific frequency and current amplitude. In the present study, we systemically examined responses of RGCs to sinusoidal electric stimulation with varying frequencies and amplitudes. We found that ON sustained alpha RGCs show distinct stimulus-response relationships to low and high frequency stimulation. For example, RGCs showed monotonic response curves to 500 Hz sinusoidal stimulation, whereas they showed non-monotonic response curves to 2000 Hz stimulation. We also described how increasing stimulus frequency gradually changed the response curves of RGCs.

Contractility and Myofibrillar Content in Skeletal Muscle are Decreased During Post-Sepsis Recovery, But Not During the Acute Phase of Sepsis.

Convalescence in humans after severe sepsis occurs over weeks to months and is associated with prolonged functional disabilities and impaired quality-adjusted survival. While much is known regarding the acute early phase of sepsis, there is a knowledge gap pertaining to restoration of muscle mass and function after elimination of the septic nidus. We used a sepsis-recovery model-where cecal-ligation-puncture (CLP) was performed in adult male mice followed 24 h later by removal of the cecum and antibiotic treatment-to assess changes in the abundance of muscle contractile proteins and function during the acute phase of sepsis (24 h post-CLP) and during the recovery phase (day 10 post-CLP). Although body weight and food consumption decreased acutely with sepsis, both had normalized by day 10; however, extensor digitorum longus mass remained decreased 10%. During acute sepsis, there were few contractile defects or significant changes in contractile proteins. In contrast, during sepsis recovery, specific maximum isometric twitch and specific maximum tetanic force were decreased ≈50%, compared with time-matched pair-fed controls, and defects were independent of the concomitant reduction in muscle mass. Force generation in sepsis-recovery mice was decreased 30% with increasing stimulus frequency. Contractile defects during sepsis-recovery were associated with 50% to 90% reductions in thin filament (troponin T, troponin I, tropomyosin, α-sarcomeric actin), thick filament (myosin heavy and myosin light chains), Z-disc (α-actinin 3), and M-band (myomesin-2) proteins, but no change in the intermediate filaments desmin and vimentin. During sepsis recovery, myofibrillar protein synthesis did not differ from control, but synthesis of sarcoplasmic proteins was increased 60%. These data suggest intrinsic defects in muscle contractile function exist during the recovery phase of sepsis and may negatively impact convalescence.

Simulating intracochlear electrocochleography with a combined model of acoustic hearing and electric current spread in the cochlea.

Intracochlear electrocochleography (ECochG) is a potential tool for the assessment of residual hearing in cochlear implant users during implantation and acoustical tuning postoperatively. It is, however, unclear how these ECochG recordings from different locations in the cochlea depend on the stimulus parameters, cochlear morphology, implant design, or hair cell degeneration. In this paper, a model is presented that simulates intracochlear ECochG recordings by combining two existing models, namely a peripheral one that simulates hair cell activation and a three-dimensional (3D) volume-conduction model of the current spread in the cochlea. The outcomes were compared to actual ECochG recordings from subjects with a cochlear implant (CI). The 3D volume conduction simulations showed that the intracochlear ECochG is a local measure of activation. Simulations showed that increasing stimulus frequency resulted in a basal shift of the peak cochlear microphonic (CM) amplitude. Increasing the stimulus level resulted in wider tuning curves as recorded along the array. Simulations with hair cell degeneration resulted in ECochG responses that resembled the recordings from the two subjects in terms of CM onset responses, higher harmonics, and the width of the tuning curve. It was concluded that the model reproduced the patterns seen in intracochlear hair cell responses recorded from CI-subjects.

The barn owls' Minimum Audible Angle.

Interaural time differences (ITD) and interaural level differences (ILD) are physical cues that enable the auditory system to pinpoint the position of a sound source in space. This ability is crucial for animal communication and predator-prey interactions. The barn owl has evolved an exceptional sense of hearing and shows abilities of sound localisation that outperform most other species. So far, behavioural studies in the barn owl often used reflexive responses to investigate aspects of sound localisation. Furthermore, they predominately probed the higher frequencies of the owl’s hearing range (> 3 kHz). In the present study we used a Go/NoGo paradigm to measure the barn owl’s behavioural sound localisation acuity (expressed as the Minimum Audible Angle, MAA) as a function of stimulus type (narrow-band noise centred at 500, 1000, 2000, 4000 and 8000 Hz, and broad-band noise) and sound source position. We found significant effects of both stimulus type and sound source position on the barn owls’ MAA. The MAA improved with increasing stimulus frequency, from 14° at 500 Hz to 6° at 8000 Hz. The smallest MAA of 4° was found for broadband noise stimuli. Comparing different sound source positions revealed smaller MAAs for frontal compared to lateral stimulus presentation, irrespective of stimulus type. These results are consistent with both the known variations in physical ITDs and variation in the width of neural ITD tuning curves with azimuth and frequency. Physical and neural characteristics combine to result in better spatial acuity for frontal compared to lateral sounds and reduced localisation acuity at lower frequencies.

Dynamic property analysis and circuit implementation of simplified memristive Hodgkin–Huxley neuron model

In this paper, the dynamic properties of a simplified memristive Hodgkin–Huxley (SMHH) neuron model are analyzed and an electric circuit is designed to completely implement it based on the mathematical equation. The sodium ion channel and potassium ion channel in the SMHH neuron model can be regarded as a second-order memristor and a first-order memristor, respectively. Under different stimuli, the SMHH neuron model can present spiking or bursting activities. In particular, when a periodic sinusoidal current is imposed on the neuron, the electrical activity behaves as bursting and it has a close frequency dependence on the external forcing current. The initiation current value of bursting activity first increases and then drops with the increase in stimulus frequency of the periodic sinusoidal current, which shows an inconsistency in traditional neural adaptation. The potential mechanism is that the frequency-dependent property of the ion channel memristors affects the threshold dynamics of the SMHH neuron. Furthermore, an analog circuit is designed to completely implement the SMHH neuron model. Since the conductance-based neuron models are mathematical complexity, only few analog circuits are designed for this kind of neuron models, which indicates that the designed circuit unit in this work will provide a great convenience to study the dynamics of conductance-based neuron and even collective behaviors of neural networks at circuit level.

Novel regulatory systems for acetylcholine release in rat striatum and anti-Alzheimer's disease drugs.

Regulation of neurotransmitter release in the central nervous system is complex. Here, we investigated regulatory mechanisms for acetylcholine (ACh) release from cholinergic neurons by performing superfusion experiments with rat striatal segments after labelling the cellular ACh pool with [3 H]choline. Electrical stimulation-evoked pronounced [3 H]ACh release from cholinergic neurons. The estimated quantity of [3 H]ACh release per pulse of electrical stimulation was reduced by an increase in stimulus frequency, showing an inverse correlation between release probability of ACh and neuronal excitation. ACh release was also negatively regulated by pre-synaptic muscarinic ACh receptors (mAChRs). The autoinhibition induced by released ACh was predominantly suppressed by the M2 -selective antagonist AF-DX 116, partially inhibited by M3 -selective darifenacin, and minimally by M4 -selective PD 102807. Other subtype-selective antagonists had no effect at subtype-selective concentrations. ACh esterase (AChE) inhibitors (diisopropylfluorophosphate, donepezil and galantamine) at concentrations that mostly inhibit esterase activity reduced [3 H]ACh release, and the reduction was abolished by treatment with atropine. This implies that pre-synaptic autoreceptors are activated more after blockade of ACh hydrolysis, leading to autoinhibition of ACh release and consequent reduction in synaptic ACh concentrations. [3 H]efflux was also enhanced by ACh uptake inhibitors (100μM hemicholinium-3 and physostigmine), regardless of ACh hydrolysis. This study shows that synaptic ACh concentrations in striatal cholinergic neurons are regulated in a complex manner by many factors such as release probability, pre-synaptic M2 /M3 /M4 mAChRs, AChE and post-synaptic ACh uptake, and provides important information about cholinergic neurotransmission for future exploration of therapeutic strategies for Alzheimer's and other central nervous system diseases. OPEN SCIENCE BADGES: This article has received a badge for *Open Materials* because it provided all relevant information to reproduce the study in the manuscript. The complete Open Science Disclosure form for this article can be found at the end of the article. More information about the Open Practices badges can be found at https://cos.io/our-services/openscience-badges/.

Auditory evoked potentials of the plainfin midshipman fish (Porichthys notatus): implications for directional hearing.

The plainfin midshipman (Porichthys notatus) is an acoustically communicative teleost fish. Here, we evaluated auditory evoked potentials (AEPs) in reproductive female midshipman exposed to tones at or near dominant frequencies of the male midshipman advertisement call. An initial series of experiments characterized AEPs at behaviorally relevant suprathreshold sound levels (130-140 dB SPL re. 1 µPa). AEPs decreased in magnitude with increasing stimulus frequency and featured a stereotyped component at twice the stimulus frequency. Recording electrode position was varied systematically and found to affect AEP magnitude and phase characteristics. Later experiments employed stimuli of a single frequency to evaluate contributions of the saccule to the AEP, with particular attention to the effects of sound source azimuth on AEP amplitude. Unilateral excision of saccular otoliths (sagittae) decreased AEP amplitude; unexpectedly, decreases differed for right versus left otolith excision. A final set of experiments manipulated the sound pressure-responsive swim bladder. Swim bladder excision further reduced the magnitude of AEP responses, effectively eliminating responses at the standard test intensity (130 dB SPL) in some animals. Higher-intensity stimulation yielded response minima at forward azimuths ipsilateral to the excised sagitta, but average cross-azimuth modulation generally remained slight. Collectively, the data underscore that electrode position is an essential variable to control in fish AEP studies and suggest that in female midshipman: (1) the saccule contributes to the AEP, but its directionality as indexed by the AEP is limited, (2) a left-right auditory asymmetry may exist and (3) the swim bladder provides gain in auditory sensitivity that may be important for advertisement call detection and phonotaxis.

The characterization of auditory brainstem response (ABR) waveforms: A study in tree shrews (Tupaia belangeri)

To characterize the patterns of ABR waves in tree shrews, we must understand the hearing sensitivity and auditory function of healthy adult tree shrews. Fifteen tree shrews (30 ears) were stimulated with clicks and tone-pips at 11 different frequencies from 1 to 60 kHz. The ABR waves were recorded and analyzed. The ABR consisted of five to seven positive waves in the first 10 ms after a click stimulus, and the average hearing threshold of component III was 27.86 ± 3.78 dB SPL. Wave III was the largest and most clear. The ABR threshold was related to the tone-pip sitmulus by a “U” shaped curve. The sensitive frequency was approximately 8 kHz in tree shrews. The latencies systematically decreased with increasing stimulus frequencies. The ABR amplitudes of wave III increased as the sound pressure level increased. All of these results provide an empirical basis for future studies of hearing diseases in tree shrews.

Tactile Frequency-Specific High-Gamma Activities in Human Primary and Secondary Somatosensory Cortices

Humans can easily detect vibrotactile stimuli up to several hundred hertz, but underlying large-scale neuronal processing mechanisms in the cortex are largely unknown. Here, we investigated the macroscopic neural correlates of various vibrotactile stimuli including artificial and naturalistic ones in human primary and secondary somatosensory cortices (S1 and S2, respectively) using electrocorticography (ECoG). We found that tactile frequency-specific high-gamma (HG, 50–140 Hz) activities are seen in both S1 and S2 with different temporal dynamics during vibration (>100 Hz). Stimulus-evoked S1 HG power, which exhibited short-delayed peaks (50–100 ms), was attenuated more quickly in vibration than in flutter (<50 Hz), and their attenuation patterns were frequency-specific within vibration range. In contrast, S2 HG power, which was activated much later than that of S1 (150–250 ms), strikingly increased with increasing stimulus frequencies in vibration range, and their changes were much greater than those in S1. Furthermore, these S1-S2 HG patterns were preserved in naturalistic stimuli such as coarse/fine textures. Our results provide persuasive evidence that S2 is critically involved in neural processing for high-frequency vibrotaction. Therefore, we propose that S1-S2 neuronal co-operation is crucial for full-range, complex vibrotactile perception in human.

Coding Properties of Three Intrinsically Distinct Retinal Ganglion Cells under Periodic Stimuli: A Computational Study.

As the sole output neurons in the retina, ganglion cells play significant roles in transforming visual information into spike trains, and then transmitting them to the higher visual centers. However, coding strategies that retinal ganglion cells (RGCs) adopt to accomplish these processes are not completely clear yet. To clarify these issues, we investigate the coding properties of three types of RGCs (repetitive spiking, tonic firing, and phasic firing) by two different measures (spike-rate and spike-latency). Model results show that for periodic stimuli, repetitive spiking RGC and tonic RGC exhibit similar spike-rate patterns. Their spike- rates decrease gradually with increased stimulus frequency, moreover, variation of stimulus amplitude would change the two RGCs' spike-rate patterns. For phasic RGC, it activates strongly at medium levels of frequency when the stimulus amplitude is low. While if high stimulus amplitude is applied, phasic RGC switches to respond strongly at low frequencies. These results suggest that stimulus amplitude is a prominent factor in regulating RGCs in encoding periodic signals. Similar conclusions can be drawn when analyzes spike-latency patterns of the three RGCs. More importantly, the above phenomena can be accurately reproduced by Hodgkin's three classes of neurons, indicating that RGCs can perform the typical three classes of firing dynamics, depending on the distinctions of ion channel densities. Consequently, model results from the three RGCs may be not specific, but can also applicable to neurons in other brain regions which exhibit part(s) or all of the Hodgkin's three excitabilities.

The Duration of Motor Responses Evoked with Intracortical Microstimulation in Rats Is Primarily Modulated by Stimulus Amplitude and Train Duration.

Microstimulation of brain tissue plays a key role in a variety of sensory prosthetics, clinical therapies and research applications, however the effects of stimulation parameters on the responses they evoke remain widely unknown. In particular, the effects of parameters when delivered in the form of a stimulus train as opposed to a single pulse are not well understood despite the prevalence of stimulus train use. We aimed to investigate the contribution of each parameter of a stimulus train to the duration of the motor responses they evoke in forelimb muscles. We used constant-current, biphasic, square wave pulse trains in acute terminal experiments under ketamine anaesthesia. Stimulation parameters were systematically tested in a pair-wise fashion in the caudal forelimb region of the motor cortex in 7 Sprague-Dawley rats while motor evoked potential (MEP) recordings from the forelimb were used to quantify the influence of each parameter in the train. Stimulus amplitude and train duration were shown to be the dominant parameters responsible for increasing the total duration of the MEP, while interphase interval had no effect. Increasing stimulus frequency from 100–200 Hz or pulse duration from 0.18–0.34 ms were also effective methods of extending response durations. Response duration was strongly correlated with peak time and amplitude. Our findings suggest that motor cortex intracortical microstimulations are often conducted at a higher frequency rate and longer train duration than necessary to evoke maximal response duration. We demonstrated that the temporal properties of the evoked response can be both predicted by certain response metrics and modulated via alterations to the stimulation signal parameters.

Effects of Direct Afferent Nerve Stimulation Frequency and Intensity In Evoking the Exercise Pressor Reflex

Contraction of freely perfused hind limb muscles in decerebrate rats evokes the exercise pressor reflex, resulting in sympathetic activation and increased blood pressure. This reflex is primarily propagated along mechanically sensitive group III and metabolically sensitive group IV afferent nerve fibers. Recent research by our laboratory has focused on the exaggeration of the exercise pressor reflex in decerebrate rats with simulated peripheral artery disease, which was induced by ligating the femoral artery for 72 hours before the start of the experiment. Recently, we showed that ligating the femoral artery increased the responses of single fiber group III and IV triceps surae muscle afferents to static contraction. The objective of the present study was to determine if electrical stimulation of group III and IV afferents at frequencies approximating those occurring during static contraction was capable of reflexively increasing arterial blood pressure.In decerebrate unanesthetized rats, we first determined motor threshold by gradually increasing the current applied as a single pulse (0.01 ms) to the gastrocnemius nerve until a twitch was evoked from the triceps surae muscles. Once motor threshold was determined, we injected intravenously pancuronium bromide to paralyze the rat for the remainder of the experiment. Five minutes later, we stimulated the sciatic nerve for 30 seconds at either 5, 20, or 100 times motor threshold, currents that presumably recruited thick group III, thick and thin group III, and all group III and IV fibers, respectively. In addition, we stimulated (0.01 ms pulse duration) the gastrocnemius nerves with six different frequencies (0.1, 0.25, 0.5, 1.0, 3.0, or 5.0 Hz), applying them in random order. Particular attention was paid to frequencies of 0.25, 0.5 and 1.0 Hz because these represented the discharge rates displayed by group III and IV afferents responding to static contraction.Overall, we found that stimulation of the gastrocnemius nerve increased arterial blood pressure in a graded fashion as both stimulus frequency and motor threshold were increased. In particular, stimulation of the gastrocnemius nerve at 100 times motor threshold (n=7) significantly increased arterial blood pressure (MAP) over baseline levels at 0.25 Hz (Peak ΔMAP= 11 ± 2 mmHg, p&lt; 0.001), 0.5 Hz (Peak ΔMAP= 13 ± 3 mmHg, p&lt; 0.01), and 1.0 Hz (Peak ΔMAP= 18 ± 4 mmHg, p&lt; 0.01).In conclusion, we observed that afferent stimulation frequencies as low as 0.25 Hz are sufficient to increase arterial blood pressure. These findings are relevant, as previous studies from our lab have measured afferent responses to contraction in these frequency ranges. Additionally, we observed that increasing stimulus frequency resulted in greater increases in blood pressure, a finding that parallels our previous results in studies using ligated rats. As such, these and future studies provide valuable insight into the mechanisms underlying the exaggerated exercise pressor reflex we observe in an animal model of peripheral artery disease.Support or Funding InformationFunded by NIH R01 ‐ AR059397

Cardiac MyBP-C regulates the rate and force of contraction in mammalian myocardium.

Cardiac myosin-binding protein-C (cMyBP-C) is a thick filament-associated protein that seems to contribute to the regulation of cardiac contraction through interactions with either myosin or actin or both. Several studies over the past several years have suggested that the interactions of cardiac myosin-binding protein-C with its binding partners vary with its phosphorylation state, binding predominantly to myosin when dephosphorylated and to actin when it is phosphorylated by protein kinase A or other kinases. Here, we summarize evidence suggesting that phosphorylation of cardiac myosin binding protein-C is a key regulator of the kinetics and amplitude of cardiac contraction during β-adrenergic stimulation and increased stimulus frequency. We propose a model for these effects via a phosphorylation-dependent regulation of the kinetics and extent of cooperative recruitment of cross bridges to the thin filament: phosphorylation of cardiac myosin binding protein-C accelerates cross bridge binding to actin, thereby accelerating recruitment and increasing the amplitude of the cardiac twitch. In contrast, enhanced lusitropy as a result of phosphorylation seems to be caused by a direct effect of phosphorylation to accelerate cross-bridge detachment rate. Depression or elimination of one or both of these processes in a disease, such as end-stage heart failure, seems to contribute to the systolic and diastolic dysfunction that characterizes the disease.

The Relationship between Frequency Dependence of Action Potential Duration and the Expression of TRPC3 in Rabbit Ventricular Myocardium

Background/Aims: Cardiac action potential duration (APD) is regulated by heart rate, leading to the trans-membrane movement of inorganic ions. Whether the alteration of heart rate can affect the expression of transient receptor potential canonical channels (TRPCs), further studies should be made. We investigated the changes of APD at different stimulus frequencies and their influences on the expression of TRPCs in rabbit ventricular myocardium. Methods: Monophasic action potential (MAP) was recorded by contact electrode technique in different programmed stimulus frequencies on rabbit ventricular epicardium in vivo, and the expression of TRPCs was detected using RT-PCR and Western blot. Results: At the frequency range of 4.5-7.5 Hz, APD gradually shortened with the increase of stimulus frequency, showing the property of significant frequency dependence in rabbit ventricular myocardium in vivo. Compared with 4.5 Hz group, TRPC3 mRNA and protein expression increased in 6 Hz and 7.5 Hz groups by way of frequency dependence. Both amiodarone (AM) and neferine (Nef) could prolongate APD and showed characters of frequency independence at the designed frequency. In contrast with 4.5 Hz control group, it was Nef treatment group rather than AM treatment group that could obviously increase the expression of TRPC3 mRNA and protein. Conclusions: At the frequency range of 4.5-7.5 Hz, frequency-dependent shortening of APD was associated with the expression of TRPC3. AM and Nef exhibited frequency-independent lengthening of the APD. Nef may prolong APD via the increasement of TRPC3.