J. Andrew Daubenspeck

Toward feedback controlled deep brain stimulation: Dynamics of glutamate release in the subthalamic nucleus in rats

Deep brain stimulation (DBS) is an effective symptomatic treatment in Parkinsons disease. High frequency stimulation (HFS) of the subthalamic nucleus elicits neurotransmitter release in multiple nuclei. Therefore, we tested the hypothesis that neurotransmitter release during HFS may be used to provide feedback control of the intensity and pattern of HFS. We studied the dynamic relationship between extracellular glutamate levels and HFS in and around the STN in anesthetized rats. We used a pseudorandom binary sequence (PRBS) of stimulation in the STN, the independent forcing function, while measuring extracellular glutamate in the same nucleus, the dependent variable. The PRBS consisted of 90 s periods during which stimulation (100 microA, 150Hz, 10% duty cycle) was either off or on. The stimulation and extracellular glutamate levels were fitted using an autoregressive exogenous model (ARX) to determine the transfer function between HFS and the extracellular glutamate concentration in the STN. The ARX model fit the dynamics of extracellular glutamate levels well (correlation coefficients ranged from 0.74 to 0.99; n=11). The transfer function accurately predicted extracellular glutamate levels in the STN even when the pattern of HFS was modified. We used the transfer function to develop a feedback controlled stimulation algorithm. Feedback controlled HFS maintained extracellular glutamate concentrations at any predefined level, but only intermittent HFS was required. We conclude that the transfer function between HFS and neurotransmitter levels in the brain can be used to design DBS protocols that generate specific temporal patterns of glutamate release in the STN.

Investigating the Contamination of Electroencephalograms by Facial Muscle Electromyographic Activity Using Matching Pursuit

It has been widely recognized and previously reported that electrical fields from facial muscle electromyographic (EMG) activity can contaminate the electroencephalogram (EEG), even when closely spaced, bipolar electrode configurations are used (personal observations). We suspected that EEG signals evoked in response to pressure changes in the upper airway may include EMG contamination subsequent to muscle reflexes triggered by the stimuli. We evaluated the potential contamination of the background EEG by voluntary activation of a facial muscle by obtaining simultaneous recordings in human subjects of the EEG (from Cz-C4) and masseter muscle EMG (from a bipolar surface electrode pair) before (quiet) and after voluntary tensing (VTen). Matching pursuit analysis permitted identification of different time-frequency patterns for each signal during the quiet period because the EMG signal has mostly atoms above 30 Hz compared to the EEG signal. However, the EEG showed periods of low-frequency activity unmatched in the EMG TF pattern below 30 Hz. During the tensing, most of the atoms of both the EEG and EMG shifted to the higher frequency regions above 100 Hz, making the separation difficult. These results further suggest that the matching pursuit method may not separate the background EEG from phasic EMG signals, both of which are nonstationary in nature.

Subtle alterations in breathing and heart rate control in the 5-HT1A receptor knockout mouse in early postnatal development

We hypothesized that absence of the 5-HT(1A) receptor would negatively affect the development of cardiorespiratory control. In conscious wild type (WT) and 5-HT(1A) receptor knockout (KO) mice, we measured resting ventilation (Ve), oxygen consumption (Vo(2)), heart rate (HR), breathing and HR variability, and the hypercapnic ventilatory response (HCVR) at postnatal day 5 (P5), day 15 (P15), and day 25 (P25). In KO mice compared with WT, we found a 17% decrease in body weight at only P5 (P < 0.01) and no effect on Vo(2). Ve was significantly (P < 0.001) lower at P5 and P25, but there was no effect on the HCVR. Breathing variability (interbreath interval), measured by standard deviation, the root mean square of the standard deviation (RMSSD), and the product of the major (L) and minor axes (T) of the Poincaré first return plot, was 57% to 187% higher only at P5 (P < 0.001). HR was 6-10% slower at P5 (P < 0.001) but 7-9% faster at P25 (P < 0.001). This correlated with changes in the spectral analysis of HR variability; the low frequency to high frequency ratio was 47% lower at P5 but 68% greater at P25. The RMSSD and (L × T) of HR variability were ~2-fold greater at P5 only (P < 0.001; P < 0.05). We conclude that 5-HT(1A) KO mice have a critical period of potential vulnerability at P5 when pups hypoventilate and have a slower respiratory frequency and HR with enhanced variability of both, suggesting abnormal maturation of cardiorespiratory control.

An inexpensive servo-respirator based upon regulation of a shunt resistance.

We have designed and built an inexpensive servo-respirator for use in investigations of respiratory control in small animals. The device uses a butterfly valve to alter the resistance of an outflow shunt from a manifold that connects the animals tracheal cannula to a pressure source. Tracheal pressure is regulated in response to a command provided by a suitably processed neural signal, often the integrated phrenic neurogram. As the valve opens, tracheal pressure approaches atmospheric; as it closes, tracheal pressure approaches the source pressure. An electronic controller circuit was developed to permit experimental procedures that include withholding volume delivery while maintaining a desired level of positive end-expiratory pressure. The device is able to track the neural command signal satisfactorily, and its performance appears to be limited primarily by the constraints applied by the respiratory system mechanics.

Temporal variation in the VT-TI relationship in humans.

The variation with time of the relationship between tidal volume (VT) and inspiratory duration (TI) was assessed by analysis of 34 breathing sequences, nominally of 300 breath duration, during eupnea and hypercapnic hyperpnea in 6 human subjects. Two approaches were used: (1) each sequence was divided into consecutive 50-breath blocks and standard regression techniques used to characterize VT as a function of TI; and (2) a piecewise linear regression technique was applied to cluster consecutive breaths into regression regimes. Analysis of covariance was used with the results of both approaches to determine the likelihood that a single regression line was adequate to describe the entire data set. Similar results obtained regardless of the approach used. In only 4 of the 34 experiments would the hypothesis of regression slope homogeneity be accepted (P greater than 0.05) using the clustering approach; in 27 experiments, it was indicated that the regression slope estimates of VT on TI should not be considered homogeneous. Changes in the VT-TI slope were not correlated with changes in mean levels of VT, TI, minute ventilation (V), mean inspiratory flow (VT/TI), nor alveolar PCO2 (PACO2). Thus it is apparent that the VT-TI relation cannot be considered temporally invariant, but changes with time over periods ranging from less than 20 to more than 100 breaths.

Grouping and multidimensional organization of respiratory sensations

Agreement has not emerged either on the sensory attributes that underlie psychophysical decisions in respiratory magnitude scaling tasks or on the physical stimuli giving rise to human judgments. Accordingly, sensory attributes of respiratory sensations were explored in the present study by adopting scaling techniques designed to help uncover the organization of sensory experiences elicited by various breathing maneuvers. Physical stimulus characteristics underlying human judgments were also considered. Subjects rated the dissimilarity between 45 pairs of breathing maneuvers on a scale ranging from 0 (“no difference at all”) to 10 (“very much difference”). Cluster analysis and multidimensional scaling were employed to uncover the perceived grouping among different types of maneuvers and the relative organization of maneuvers in a Euclidean space. Dissimilarity judgments varied primarily as a function of a single attribute, degree or magnitude of external loading, ranging from an infinite respiratory load to no load at all. This continuum may depend on interactions among pressure or tension, airflow, and lung volume displacement. A secondary attribute may relate to airflow direction (inspiration or expiration).

Effects of non-REM sleep upon respiratory drive and the respiratory pump in humans.

We observed that ventilation fell and end-tidal CO2 rose in the change from wakefulness to non-REM (NREM) sleep in 4 normal human subjects studied on two nights each. We hypothesized that the observed ventilatory depression was due to effects of sleep both upon the central respiratory neural output and upon the mechanical respiratory pump. Both the central controller response to CO2, as measured by diaphragmatic and intercostal EMG activity, and the ability of the respiratory pump in effecting ventilation in response to diaphragmatic or intercostal activation, as measured by the relationship between the EMG activities and minute ventilation, are reduced in NREM sleep. We describe a general method of apportioning the separate effects of sleep, or other factors, upon the central respiratory controller, the respiratory mechanical pump, and the metabolic rate, in determining the total observed increase in end-tidal CO2.

Expiratory pattern and laryngeal responses to single-breath expiratory resistance loads

Responses of expiratory duration (TE) and laryngeal aperture to small flow resistance loads (2 and 5 cm H2O/LPS) applied to single expirations were measured using repeated applications in four subjects during quiet breathing. All subjects significantly prolonged TE in response to the higher load and 3 of the 4 showed that same response to the lower load, which was not perceived by these subjects. These same 3 subjects showed a narrowed laryngeal aperture in response to loading such that their expiratory impedance must have been greater than the increase provided by the load alone. The effect of such a loading response was to slow the expiratory volume decay so that a small but significant increase in the halftime for volume decay was observed. The prolongation of TE seen with loading could be due to the alteration of volume-related feedback consequent to the increased expiratory impedance. This may serve a role in regulation of expiratory muscle function.

Partial Raphe Dysfunction in Neurotransmission Is Sufficient to Increase Mortality after Anoxic Exposures in Mice at a Critical Period in Postnatal Development

Sudden infant death syndrome (SIDS) cases often have abnormalities of the brainstem raphe serotonergic (5-HT) system. We hypothesize that raphe dysfunction contributes to a failure to autoresuscitate from multiple hypoxic events, leading to SIDS. We studied autoresuscitation in two transgenic mouse models in which exocytic neurotransmitter release was impaired via conditional expression of the light chain from tetanus toxin (tox) in raphe neurons expressing serotonergic bacterial artificial chromosome drivers Pet1 or Slc6a4. These used recombinase drivers targeted different portions of medullary raphe serotonergic, tryptophan hydroxylase 2 (Tph2)+ neurons by postnatal day (P) 5 through P12: approximately one-third in triple transgenic Pet1::Flpe, hβactin::cre, RC::PFtox mice; approximately three-fourths in Slc6a4::cre, RC::Ptox mice; with the first model capturing a near equal number of Pet1+,Tph2+ versus Pet1+,Tph2low or negative raphe cells. At P5, P8, and P12, “silenced” mice and controls were exposed to five, ∼37 s bouts of anoxia. Mortality was 5–10 times greater in “silenced” pups compared with controls at P5 and P8 (p = 0.001) but not P12, with cumulative survival not differing between experimental transgenic models. “Silenced” pups that eventually died took longer to initiate gasping (p = 0.0001), recover heart rate (p = 0.0001), and recover eupneic breathing (p = 0.011) during the initial anoxic challenges. Variability indices for baseline breathing distinguished “silenced” from controls but did not predict mortality. We conclude that dysfunction of even a portion of the raphe, as observed in many SIDS cases, can impair ability to autoresuscitate at critical periods in postnatal development and that baseline indices of breathing variability can identify mice at risk. SIGNIFICANCE STATEMENT Many sudden infant death syndrome (SIDS) cases exhibit a partial (∼26%) brainstem serotonin deficiency. Using recombinase drivers, we targeted different fractions of serotonergic and raphe neurons in mice for tetanus toxin light chain expression, which prevented vesicular neurotransmitter release. In one model, approximately one-third of medullary Tph2+ neurons are silenced by postnatal (P) days 5 and 12, along with some Pet1+,Tph2low or negative raphe cells; in the other, approximately three-fourths of medullary Tph2+ neurons, also with some Tph2low or negative cells. Both models demonstrated excessive mortality to anoxia (a postulated SIDS stressor) at P5 and P8. We demonstrated fatal vulnerability to anoxic stress at a specific time in postnatal life induced by a partial defect in raphe function. This models features of SIDS.

Acute hypoxia activates human 8–12 Hz physiological tremor

Hypoxia causes arousal. Therefore, we hypothesized that hypoxia activates the human somatomotor system and should augment tremor. We determined the effects of hypoxia, PET(O2) = 45+/-2.2 mm Hg, hypocapnia, and the hypocapnic-hypoxic interaction on finger tremor during elastic loading. A total of 12 healthy male volunteers were studied during five conditions: eupnea, hypocapnic hypoxia, eucapnic hypoxia, hypocapnic normoxia, and eucapnic normoxia. Acceleration power spectra were computed to quantify 8-12 Hz tremor. Hypoxia significantly augmented 8-12 Hz physiological tremor (P=0.002). Furthermore, six subjects (50%) exhibited significantly more tremor during hypocapnic hypoxia (hH) than during eucapnic hypoxia (eH). We conclude that acute hypoxia augments 8-12 Hz physiological tremor, and hypocapnia further augments this tremor in some subjects. As such, hypoxic tremor is activated physiological tremor, and entrainment of spinal alpha-motoneuron activity may be the final common pathway.