This study aims to investigate the evoked and the induced activity in 40 Hz auditory responses. The 40 Hz activity, also called Pb or P50 or P1 component, has a latency of 50ms and belongs to the category of MLRs (Middle latency responses), which occur right after Auditory Brainstem Responses (ABRs) between 15 and 80ms. Its importance is related to possible clinical applications such as anesthesia, schizophrenia and auditory development. In addition to this, evoked and induced activities at 40 Hz might play an important role in cognitive processing. Trains of right ear rarefaction clicks at the mean rate of 39.1 Hz, intensity of 50dB and duration of 100 were used to elicit the above mentioned activities. Three different sequences have been used: steady state, low jittered and medium jittered. Low jittered sequence has been the main sequence used to study the 40 Hz activity. The advantage of using this sequence is the fact that a deconvolution analysis can be performed and also the fact that it does not differ too much from the standard 40Hz steady-state sequence and this means that a resonance at 40 Hz can still be obtained. Ten healthy subjects (8 males and 2 females; ages ranging from 25 to 47), with no history of audiological or neurological hearing impairment were recorded. Informed consent was signed according to approved IRB protocols. All recordings were done in a sound-proof chamber (Acoustic Systems, Inc.) with subjects lying on a bed comfortably. The subjects were not asked to perform any tasks, but just to passively listen to the acoustical stimuli. Evoked and induced activities were recorded in response to the above mentioned acoustic stimuli. The deconvolution analysis showed that the peak of activity occurs around 152ms. Wavelets analysis has confirmed this observation and has also unveiled and induced activity in the low beta range. This induced activity seems to be strictly related to the evoked activity, as it seems to occur around 390ms, which corresponds to the situation where the 40 Hz evoked activity enters a steady state condition, which lasts until the last acoustic stimulus has been applied. The latter observation is again in accordance with the literature, where it is reported that the 40Hz evoked activity could reflect the initial coactivation of neural assemblies representing specific stimulus features. A change in such stimulus features could be reflected as induced oscillations occurring in the middle beta range (16-22 Hz).

Recent research shows that human auditory steady-state responses (ASSRs) develop a resonance at 40 Hz and the dramatic amplitude increase of the Pb component of the middle latency response (MLR) accounts for the high amplitude of the ASSR at 40 Hz. The first part of this study aimed to investigate the ASSR resonance characteristics as a function of rate in guinea pigs. A study of the grand average of the peak-to-peak and fundamental frequency amplitudes does indeed show a resonance around 40 Hz in guinea pigs. Unlike human ASSRs, this resonance is very broad (26-52 Hz) and flat. The centrally recorded ASSRs are smaller and tend to have resonances at higher rates compared to temporal signals. The second part of the analysis investigated whether the superposition of transient responses can predict the acquired ASSRs at each corresponding rate. This superposition theory is one of two competing theories on the origin of the ASSRs, with the other centering on the induced phase synchronization of brain waves. In order to test the first theory, transient responses were used to create synthetic ASSRs, which were then compared to the acquired ASSRs via correlation coefficient and phasor analysis. For the 40 Hz ASSR, both temporal and central electrode synthesized ASSRs show a correlation coefficient above 0.80. In the comparison at 20 Hz, the correlation coefficient is very high (about 0.9) in the temporal electrode, yet significantly lower (about 0.7) for the central electrode. Furthermore, at 80 Hz, the correlation coefficient is significantly lower in both temporal and central electrodes (about 0.7). At all rates, the correlation coefficients are highest with low jitter sequences. Finally, phasor analysis was also used to test the superposition theory of the generation of the acquired ASSRs at 20, 40, and 80 Hz. Overall, in the temporal recordings at 40 Hz, the superposition of the MLR responses accurately predicted the acquired 40 Hz ASSR as demonstrated by both magnitude and phase analysis. The recordings made in the central electrode only predicted the acquired ASSR in its phases, with significant differences found in magnitude at its main harmonics. Similarly, at 20 and 80 Hz in both temporal and central electrodes, the synthetic ASSRs did not appear to fully predict the acquired ASSRs. Although the phases were successfully predicted, large magnitude variations were observed. As shown by mean prediction error plots, the acquired ASSRs are best predicted by low jitter sequences, followed by low-medium and medium jitter sequences.

Oscillations in the brain’s electrical potential, recorded through the technique of electroencephalography (EEG), reflect the ensemble activity of a large population of neurons. Auditory steady-state response (ASSR) is the time-locked entrainment in EEG to an auditory stimulus such as a train of clicks. ASSR to a 40 Hz (gamma frequency) click train is especially reduced in schizophrenia patients, reflecting the sensory processing deficits that impact real-world functional outcomes. Since the 40 Hz ASSR is demonstrable across species and is responsive to pharmacological treatments, it can be a translational biomarker for drug development studies. Prototypical antipsychotic drugs (APDs) like haloperidol and clozapine are examples of typical and atypical classes used to treat schizophrenia patients. While both are D2 receptor blockers, they have additional pharmacological effects that may differentiate them. Here, we investigated the acute effect of clozapine (atypical) or haloperidol (typical) on the 40 Hz ASSR, in two independent studies. The doses for the two drugs were chosen to reflect comparable in vivo D2 receptor occupancy. We used female Sprague-Dawley rats implanted with epidural EEG recording electrodes. In the first experiment, vehicle or clozapine 2.5, 5, and 10 mg/kg were administered (sc) and the 40 Hz ASSR paradigm (65 dB, 40 clicks for 1 second, 2-sec inter-stimulus interval) was used to record responses at 30, 60, 90 and 120 minutes post-drug. Resting-state EEG was recorded at 60 minutes post-treatment. Treatment effects were evaluated on the evoked power and phase-locking factor (PLF), a measure of trial-to-trial consistency of the 40 Hz ASSR. Clozapine improved both measures in a dose and time-dependent manner. Clozapine also tended to reduce the resting-state gamma (30-100 Hz) power, a hallmark of cortical noise. However, the effect was not significant (P>0.05). Next, we tested the effect of haloperidol on the 40 Hz ASSR. Doses of 0.02 mg/kg -0.08 mg/kg (sc) were evaluated at 30, 60, 90 and 120-minutes post-injection. Haloperidol failed to improve the 40 Hz ASSR (evoked power and PLF). Moreover, it had no discernible effect on the resting-state gamma. These results show that despite the comparable blockade of D2 receptors, the putative target for these APDs, clozapine, and haloperidol have different effects on the 40 Hz ASSR. We conclude that the effects of clozapine on 40 Hz ASSR may be unrelated to its affinity to D2 receptors and may be mediated through other pharmacological mechanisms.

The 40 Hz auditory steady state response (ASSR) is an EEG response of local neural synchrony that is evoked by the repeated presentation of a 40 Hz click train. While the principal cortical generators of this response appear to be the bilateral primary auditory cortices as they show the largest phase locking and evoked power, other regions across the cortical mantle synchronize too, including the prefrontal cortex (PFC) that receives input from the primary auditory cortex and is involved in higher order cognitive functions. In schizophrenia, it is hypothesized that NMDA-mediated disruption in PFC function contributes to cognitive deficits including working memory and executive function. In rodents, NMDA antagonists reliably disrupt set shifting, a working memory task linked to PFC function. It is however not known if NMDA antagonism would disrupt the 40 Hz ASSR in PFC. In the following study, we equipped a group of female SD rats with epidural electrodes targeting the PFC (2.5 mm anterior and 0.8 mm lateral to bregma) and the primary auditory cortex (4.5 mm caudal, 7.5 mm lateral and 3.5 mm ventral to bregma). Two epidural screw electrodes on cerebellum served as ground and reference. After recovery from surgery and acclimation, rats were pretreated with small to modest doses of the NMDA antagonist MK801 (0.025, 0.05 and 0.1 mpk) or saline (1 ml/kg, sc) in a cross-over design, tethered to EEG cables and the EEG signal was amplified and acquired (Signal 7.0; CED1401 Micro 3). Trains of square waves (~ 1 ms duration; 40/s) were generated and played through the house speakers at ~ 65 dB SPL. EEG was acquired as 4 s sweeps while the click train played between 1-2 s of each sweep; 75 trials were recorded from each subject. Sixty minutes after vehicle treatment, robust EEG entrainment was noted in both the temporal cortex as well as the PFC. As expected, the EEG signal power from the temporal cortex was notably larger compared to the PFC. Nevertheless, both regions showed clear 40 Hz entrainment to click trains. However, MK801 effect on the 40 Hz ASSR was disparate across the two regions. In the prefrontal cortex, the intertrial coherence (ITC) of the 40 Hz ASSR was strongly disrupted by MK801 at all doses (P<0.001; Dunnett’s test). Evoked power was significantly reduced only at the highest dose (P<0.0001). In primary auditory cortex, relative to vehicle treatment, evoked power showed a significant increase after 0.025 mpk and 0.05 mpk dose but declined significantly after the 0.1 mpk dose (P<0.05). However, ITC was unaffected (P>0.05). These results indicate that gamma neural synchrony in the PFC is more vulnerable to NMDA antagonist- mediated disruption, as compared to the primary auditory cortex. Moreover, it suggests that executive and cognitive functions may be more readily compromised by NMDA-mediated transmission disturbance even as auditory processing is enhanced or unaffected.

Schizophrenia is a troubling and severe mental illness that is only incompletely treated by currently available drugs. New drug development is hindered by a scarcity of functionally relevant pharmacodynamic biomarkers that are translatable across preclinical and human subjects. Although psychosis is a major feature of schizophrenia, cognitive and negative symptoms determine the long-term functional outcomes for patients. Stimulus-evoked neural synchrony at gamma (~ 40 Hz) frequency plays an important role in the processing and integration of sensory information. Not surprisingly, schizophrenia patients show deficits in gamma oscillations. NMDA receptor (NMDAR) activation on fast-spiking parvalbumin-positive interneurons is deemed important for the generation of gamma oscillations. NMDA hypofunction has been proposed as an alternative hypothesis to the well-known dopamine dysregulation to explain the neurochemical abnormalities associated with schizophrenia. For this dissertation, we validated a preclinical model to pharmacologically probe NMDA-mediated gamma oscillations by further characterizing the auditory-steady state response (ASSR) in female Sprague Dawley rats. The ASSR is a measure of cortical neural synchrony evoked in response to periodic auditory stimuli. ASSR at 40 Hz is consistently disrupted in patients. First, we established the reliability of click train-evoked 40 Hz ASSR and tone-evoked gamma oscillations in 6 separate sessions, spread over a 3-week period. Then we established the sensitivity of these neural synchrony measures to acute NMDAR blockade using the high affinity NMDA channel blocker MK-801, using a repeated measures design. Next, we compared the reliability and sensitivity of the 40 Hz ASSR from two distinct recording sites. Results from this study showed that as compared to vertex, temporal recording showed a greater gamma synchrony. However, the temporal recording had poor test-retest reliability and lower sensitivity to MK-801-induced disruption. Lastly, we characterized the dose-response profiles of an NMDA co-agonist D-serine, an atypical (clozapine) and a typical (haloperidol) antipsychotic, on the 40 Hz ASSR. Results from these studies showed that only clozapine was effective in robustly augmenting 40 Hz ASSR. Furthermore, only clozapine pretreatment had partial protective effect against MK-801 induced ASSR disruption. Overall, this work establishes that vertex recorded 40 Hz ASSR is a reliable neural synchrony biomarker in female SD rats that is amenable for bidirectional pharmacodynamic modulation.

Acute mental fatigue can negatively affect cognitive performance This study investigates how different auditory interventions can influence the degree of mental fatigue during a cognitively demanding task, in this case, a 16-minute T-LoadDback test designed to induce mental fatigue. 27 healthy individuals were recruited and randomly assigned to one of three groups. One group listened to self-selected music, as it is well documented that listening to preferred music can increase dopamine levels, potentially mitigating mental fatigue. Another group listened to a 40 Hz binaural beat, known to improve concentration and increase energy. The third group served as a control group and conducted the task in silence. Subjective measurements of mental fatigue were obtained before and after the T-LoadDback test using a Visual Analog Scale for Fatigue(VAS-F), while objective data on mental fatigue were collected through the results of a 5-minute Corsi Block-tapping test and heart rate variability(HRV), both measured before and after the main task. The results indicated no significant pre-post differences for the VAS-F scores, Corsi test performance, or HRV, suggesting that mental fatigue was not successfully induced in the current experiment.The VAS-F scores showed no significant main effects or group differences, despite notable individual variations. Although both the music and 40 Hz groups outperformed the control group in the Corsi test, no differences were observed between the intervention groups themselves.  Similarly, HRV data showed no significant changes, indicating minimal effects on autonomic nervous system activity. These results suggest that while individual preferences in auditory stimuli, such as ambient music, might influence fatigue levels, the overall effectiveness of such interventions remains inconclusive.