The motor apparatus of head movements in the Oleander hawkmoth (Daphnis nerii, Lepidoptera).

Head movements of insects play a vital role in diverse locomotory behaviors including flying and walking. Because insect eyes move minimally within their sockets, their head movements are essential to reduce visual blur and maintain a stable gaze. As in most vertebrates, gaze stabilization behavior in insects requires the integration of both visual and mechanosensory feedback by the neck motor neurons. Although visual feedback is derived from the optic flow over the retina of their compound eyes, mechanosensory feedback is derived from their organs of balance, similar to the vestibular system in vertebrates. In Diptera, vestibular feedback is derived from the halteres-modified hindwings that evolved into mechanosensory organs-and is integrated with visual feedback to actuate compensatory head movements. However, non-Dipteran insects, including Lepidoptera, lack halteres. In these insects, vestibular feedback is obtained from the antennal Johnston's organs but it is not well-understood how it integrates with visual feedback during head movements. Indeed, although head movements are well-studied in flies, the underlying motor apparatus in non-Dipteran taxa has received relatively less attention. As a first step toward understanding compensatory head movements in the Oleander hawkmoth Daphnis nerii, we image the anatomy and architecture of their neck joint sclerites and muscles using X-ray microtomography, and the associated motor neurons using fluorescent dye fills and confocal microscopy. Based on these morphological data, we propose testable hypotheses about the putative function of specific neck muscles during head movements, which can shed light on their role in neck movements and gaze stabilization.

Cerebellar control of a unitary head direction sense.

The head-direction (HD) system, a key neural circuit for navigation, consists of several anatomical structures containing neurons selective to the animal's head direction. HD cells exhibit ubiquitous temporal coordination across brain regions, independently of the animal's behavioral state or sensory inputs. Such temporal coordination mediates a single, stable, and persistent HD signal, which is essential for intact orientation. However, the mechanistic processes behind the temporal organization of HD cells are unknown. By manipulating the cerebellum, we identify pairs of HD cells recorded from two brain structures (anterodorsal thalamus and retrosplenial cortex) that lose their temporal coordination, specifically during the removal of the external sensory inputs. Further, we identify distinct cerebellar mechanisms that participate in the spatial stability of the HD signal depending on sensory signals. We show that while cerebellar protein phosphatase 2B-dependent mechanisms facilitate the anchoring of the HD signal on the external cues, the cerebellar protein kinase C-dependent mechanisms are required for the stability of the HD signal by self-motion cues. These results indicate that the cerebellum contributes to the preservation of a single and stable sense of direction.

The use of transcutaneous Vagal Nerve Stimulation as an effective countermeasure for Simulator Adaptation Syndrome.

This research focused on investigating the effectiveness of Transcutaneous Vagal Nerve Stimulation (tVNS) as compared to Galvanic Cutaneous Stimulation (GCS) at mitigating Simulator Adaptation Syndrome (SAS). Fifty drivers (mean age = 23.04 ± 17.71 years old, twenty-two men) participated in a driving simulation experiment. The total scores of the Simulator Sickness Questionnaire, head movements (body balance index), and driving performance variables were measured under five stimulation conditions: i) baseline (no stimulation delivered), ii) sham GCS, iii) sham tVNS, iv) active GCS, and v) active tNVS. The results showed that tVNS alleviated SAS and improved driving performance variables more effectively than GCS. We conclude that GCS and tVNS have similar neurological mechanisms to reduce SAS, providing possible explanations for the greater effectiveness of tVNS. We encourage the use of tVNS to decrease SAS.

MgADP Promotes Myosin Head Movement toward Actin at Low [Ca2+] to Increase Force Production and Ca2+-Sensitivity of Contraction in Permeabilized Porcine Myocardial Strips.

Myosin cross-bridges dissociate from actin following Mg2+-adenosine triphosphate (MgATP) binding. Myosin hydrolyses MgATP into inorganic phosphate (Pi) and Mg2+-adenosine diphosphate (ADP), and release of these hydrolysis products drives chemo-mechanical energy transitions within the cross-bridge cycle to power muscle contraction. Some forms of heart disease are associated with metabolic or enzymatic dysregulation of the MgATP-MgADP nucleotide pool, resulting in elevated cytosolic [MgADP] and impaired muscle relaxation. We investigated the mechanical and structural effects of increasing [MgADP] in permeabilized myocardial strips from porcine left ventricle samples. Sarcomere length was set to 2.0 µm at 28 °C, and all solutions contained 3% dextran T-500 to compress myofilament lattice spacing to near-physiological values. Under relaxing low [Ca2+] conditions (pCa 8.0, where pCa = -log10[Ca2+]), tension increased as [MgADP] increased from 0-5 mM. Complementary small-angle X-ray diffraction measurements show that the equatorial intensity ratio, I1,1/I1,0, also increased as [MgADP] increased from 0 to 5 mM, indicating myosin head movement away from the thick-filament backbone towards the thin-filament. Ca2+-activated force-pCa measurements show that Ca2+-sensitivity of contraction increased with 5 mM MgADP, compared to 0 mM MgADP. These data show that MgADP augments tension at low [Ca2+] and Ca2+-sensitivity of contraction, suggesting that MgADP destabilizes the quasi-helically ordered myosin OFF state, thereby shifting the cross-bridge population towards the disordered myosin ON state. Together, these results indicate that MgADP enhances the probability of cross-bridge binding to actin due to enhancement of both thick and thin filament-based activation mechanisms.

Neck Strength in Coronal and Sagittal Head Movement in Military Pilots and Aircrew With Chronic Neck Pain: A Systematic Review With Meta-Analysis.

OBJECTIVE: This systematic review aimed to assess whether there is a difference in neck strength between military pilots and aircrew with and without chronic neck pain. METHODS: The PubMed, Embase, and Scopus databases were searched. Two independent reviewers selected relevant full articles comparing neck strength between military pilots and aircrew with and without chronic neck pain. Two independent reviewers extracted the data from the full articles selected. A meta-analysis was used to assess standardized mean differences in neck strength based on a random-effects model. RESULTS: The search returned 3554 results; 5 articles were included in the study. Military pilots and aircrew with chronic neck pain showed no difference in neck strength from military pilots and aircrew without chronic neck pain for flexion and extension, but did show a neck strength reduction for right and left lateral flexion -0.29 (95% confidence interval, -0.52 to -0.06; I² = 3%) and -0.23 (95% confidence interval, -0.45 to 0.00; I² = 0%), respectively. CONCLUSION: Based on this meta-analysis with a 3a level of evidence, military pilots and aircrew with chronic neck pain have reduced neck strength for coronal head movement, but not sagittal movement compared with military pilots and aircrew without chronic neck pain.

The Neural Correlates of Spatial Disorientation in Head Direction Cells.

While the brain has evolved robust mechanisms to counter spatial disorientation, their neural underpinnings remain unknown. To explore these underpinnings, we monitored the activity of anterodorsal thalamic head direction (HD) cells in rats while they underwent unidirectional or bidirectional rotation at different speeds and under different conditions (light vs dark, freely-moving vs head-fixed). Under conditions that promoted disorientation, HD cells did not become quiescent but continued to fire, although their firing was no longer direction specific. Peak firing rates, burst frequency, and directionality all decreased linearly with rotation speed, consistent with previous experiments where rats were inverted or climbed walls/ceilings in zero gravity. However, access to visual landmarks spared the stability of preferred firing directions (PFDs), indicating that visual landmarks provide a stabilizing signal to the HD system while vestibular input likely maintains direction-specific firing. In addition, we found evidence that the HD system underestimated angular velocity at the beginning of head-fixed rotations, consistent with the finding that humans often underestimate rotations. When head-fixed rotations in the dark were terminated HD cells fired in bursts that matched the frequency of rotation. This postrotational bursting shared several striking similarities with postrotational "nystagmus" in the vestibulo-ocular system, consistent with the interpretation that the HD system receives input from a vestibular velocity storage mechanism that works to reduce spatial disorientation following rotation. Thus, the brain overcomes spatial disorientation through multisensory integration of different motor-sensory inputs.

Removal of physiological artifacts from simultaneous EEG and fMRI recordings.

OBJECTIVE: Simultaneous recording of the electroencephalogram (EEG) and functional magnetic resonance imaging (fMRI) allows a combination of eletrophysiological and haemodynamic information to be used to form a more complete picture of cerebral dynamics. However, EEG recorded within the MRI scanner is contaminated by both imaging artifacts and physiological artifacts. The majority of the techniques used to pre-process such EEG focus on removal of the imaging and balistocardiogram artifacts, with some success, but don't remove all other physiological artifacts. METHODS: We propose a new offline EEG artifact removal method based upon a combination of independent component analysis and fMRI-based head movement estimation to aid the removal of physiological artifacts from EEG recorded during EEG-fMRI recordings. Our method makes novel use of head movement trajectories estimated from the fMRI recording in order to assist with identifying physiological artifacts in the EEG and is designed to be used after removal of the fMRI imaging artifact from the EEG. RESULTS: We evaluate our method on EEG recorded during a joint EEG-fMRI session from healthy adult participants. Our method significantly reduces the influence of all types of physiological artifacts on the EEG. We also compare our method with a state-of-the-art physiological artifact removal method and demonstrate superior performance removing physiological artifacts. CONCLUSIONS: Our proposed method is able to remove significantly more physiological artifact components from the EEG, recorded during a joint EEG-fMRI session, than other state-of-the-art methods. SIGNIFICANCE: Our proposed method represents a marked improvement over current processing pipelines for removing physiological noise from EEG recorded during a joint EEG-fMRI session.

Cervical Muscle Activation Due to an Applied Force in Response to Different Types of Acoustic Warnings.

Mild traumatic brain injury (mTBI) and whiplash-associated disorder are the most common head and neck injuries and result from a sudden head or body acceleration. The head and neck injury potential is correlated with the awareness, level of muscle activation, and posture changes at the time of the perturbation. Environmental acoustic stimuli or a warning system can influence muscle activation and posture during a head perturbation. In this study, different acoustic stimuli, including Non-Directional, Directional, and Startle, were provided 1000 ms before a head impact, and the amplitude and timing of cervical muscle electromyographic (EMG) data were characterized based on the type of warning. The startle warning resulted in 49% faster and 80% greater EMG amplitude compared to the Directional and Non-Directional warnings after warning and before the impact. The post-impact peak EMG amplitudes in Unwarned trials were lower by 18 and 21% in the retraction and rebound muscle groups, respectively, compared to any of the warned conditions. When there was no warning before the impact, the retraction and rebound muscle groups also reached their maximum activation 38 and 54 ms sooner, respectively, compared to the warned trials. Based on these results, the intensity and complexity of information that a warning sound carries change the muscle response before and after a head impact and has implications for injury potential.

Effectiveness of pediatric integrative manual therapy in cervical movement limitation in infants with positional plagiocephaly: a randomized controlled trial.

BACKGROUND: Positional plagiocephaly (PP) is a cranial deformation frequent amongst children and consisting in a flattened and asymmetrical head shape. PP is associated with excessive time in supine and with congenital muscular torticollis (CMT). Few studies have evaluated the efficiency of a manual therapy approach in PP. The purpose of this parallel randomized controlled trial is to compare the effectiveness of adding a manual therapy approach to a caregiver education program focusing on active rotation range of motion (AROM) and neuromotor development in a PP pediatric sample. METHODS: Thirty-four children with PP and less than 28 week-old were randomly distributed into two groups. AROM and neuromotor development with Alberta Infant Motor Scale (AIMS) were measured. The evaluation was performed by an examiner, blinded to the randomization of the subjects. A pediatric integrative manual therapy (PIMT) group received 10-sessions involving manual therapy and a caregiver education program. Manual therapy was addressed to the upper cervical spine to mobilize the occiput, atlas and axis. The caregiver educational program consisted in exercises to reduce the positional preference and to stimulate motor development. The control group received the caregiver education program exclusively. To compare intervention effectiveness across the groups, improvement indexes of AROM and AIMS were calculated using the difference of the final measurement values minus the baseline measurement values. If the distribution was normal, the improvement indexes were compared using the Student t-test for independent samples; if not, the Mann-Whitney U test was used. The effect size of the interventions was calculated using Cohen's d. RESULTS: All randomized subjects were analysed. After the intervention, the PIMT group showed a significantly higher increase in rotation (29.68 ± 18.41°) than the control group (6.13 ± 17.69°) (p = 0.001). Both groups improved the neuromotor development but no statistically significant differences were found. No harm was reported during the study. CONCLUSION: The PIMT intervention program was more effective in increasing AROM than using only a caregiver education program. The study has been retrospectively registered at clinicaltrials.gov, with identification number NCT03659032 . Registration date: September 1, 2018.

DeepLabStream enables closed-loop behavioral experiments using deep learning-based markerless, real-time posture detection.

In general, animal behavior can be described as the neuronal-driven sequence of reoccurring postures through time. Most of the available current technologies focus on offline pose estimation with high spatiotemporal resolution. However, to correlate behavior with neuronal activity it is often necessary to detect and react online to behavioral expressions. Here we present DeepLabStream, a versatile closed-loop tool providing real-time pose estimation to deliver posture dependent stimulations. DeepLabStream has a temporal resolution in the millisecond range, can utilize different input, as well as output devices and can be tailored to multiple experimental designs. We employ DeepLabStream to semi-autonomously run a second-order olfactory conditioning task with freely moving mice and optogenetically label neuronal ensembles active during specific head directions.

Joint and individual effectiveness of galvanic cutaneous stimulation and tactile stimulation at decreasing Simulator Adaptation Syndrome.

This research was focused on investigating the effectiveness of galvanic cutaneous stimulation and tactile stimulation jointly and individually at mitigating Simulator Adaptation Syndrome. Forty drivers (mean age = 23.1 ± 3.4 years old, twenty women) participated in a driving simulation experiment. Total scores of the Simulator Sickness Questionnaire, head movements (an index of body balance), and driving performance variables were compared across four different stimulation conditions: i) baseline (where no stimulation was presented), ii) galvanic cutaneous stimulation and iii) tactile stimulation deployed individually, and iv) both techniques deployed jointly. The results showed that both techniques presented in conjunction alleviate Simulator Adaptation Syndrome and improve driving performance more effectively than when they are presented in isolation. Importantly, reduced head movements were only revealed when galvanic cutaneous stimulation was applied. We concluded that the reduction of this syndrome is due to an improvement of body balance (elicited by galvanic cutaneous stimulation), and a distraction from the symptoms (elicited by tactile stimulation). We encourage the use of both techniques simultaneously to decrease Simulator Adaptation Syndrome.

A Pilot Longitudinal Study of 3-Dimensional Head and Neck Kinematics During Functional Tasks in Individuals With Chronic Idiopathic Neck Pain Either Wait-Listed for or Receiving Chiropractic Spinal Manipulative Therapy With Exercise.

OBJECTIVE: The purpose of this study was to determine if there is a relationship between pain and movement kinematics during functional tasks, evaluated over time, in individuals with chronic idiopathic neck pain. METHODS: Ten participants with chronic idiopathic neck pain performed 2 functional tasks (overhead reach to the right and putting on a seatbelt) while evaluated using 8 Oqus 300+ cameras. Kinematic variables included joint angles and range of motion (ROM) (°), head segment relative to neck segment (head-neck [HN]); and head/neck segment relative to upper thoracic segment (head/neck-trunk), velocity (m/s), and time (% of movement phase). Pain was quantified using a 100-mm visual analog scale. Linear mixed effects regression models were used to analyze associations between pain and kinematic variables adjusting for treatment group. RESULTS: For overhead reach, higher pain was associated with less HN peak rotation at baseline (ß = -0.33; 95% CI -0.52 to -0.14, P = .003) and less HN total rotation ROM at 6 months (ß = -0.19; 95% CI -0.38 to -0.003, P = .048). For the seatbelt task, higher pain was associated with less HN peak rotation (ß = -0.52; 95% CI -0.74 to -0.30 to -0.74, P < .001) and less HN total rotation ROM at baseline (ß = -0.32; 95% CI -0.53 to -0.10, P = .006). No other movement variables demonstrated meaningful relationships with pain for the reach or seatbelt tasks. CONCLUSION: Higher pain is associated with less HN peak and total rotation during functional reaching tasks requiring head rotation. Recognizing altered functional kinematics in individuals with chronic neck pain may assist patient management.

Automated detection of the head-twitch response using wavelet scalograms and a deep convolutional neural network.

Hallucinogens induce the head-twitch response (HTR), a rapid reciprocal head movement, in mice. Although head twitches are usually identified by direct observation, they can also be assessed using a head-mounted magnet and a magnetometer. Procedures have been developed to automate the analysis of magnetometer recordings by detecting events that match the frequency, duration, and amplitude of the HTR. However, there is considerable variability in the features of head twitches, and behaviors such as jumping have similar characteristics, reducing the reliability of these methods. We have developed an automated method that can detect head twitches unambiguously, without relying on features in the amplitude-time domain. To detect the behavior, events are transformed into a visual representation in the time-frequency domain (a scalogram), deep features are extracted using the pretrained convolutional neural network (CNN) ResNet-50, and then the images are classified using a Support Vector Machine (SVM) algorithm. These procedures were used to analyze recordings from 237 mice containing 11,312 HTR. After transformation to scalograms, the multistage CNN-SVM approach detected 11,244 (99.4%) of the HTR. The procedures were insensitive to other behaviors, including jumping and seizures. Deep learning based on scalograms can be used to automate HTR detection with robust sensitivity and reliability.

Automated task training and longitudinal monitoring of mouse mesoscale cortical circuits using home cages.

We report improved automated open-source methodology for head-fixed mesoscale cortical imaging and/or behavioral training of home cage mice using Raspberry Pi-based hardware. Staged partial and probabilistic restraint allows mice to adjust to self-initiated headfixation over 3 weeks' time with ~50% participation rate. We support a cue-based behavioral licking task monitored by a capacitive touch-sensor water spout. While automatically head-fixed, we acquire spontaneous, movement-triggered, or licking task-evoked GCaMP6 cortical signals. An analysis pipeline marked both behavioral events, as well as analyzed brain fluorescence signals as they relate to spontaneous and/or task-evoked behavioral activity. Mice were trained to suppress licking and wait for cues that marked the delivery of water. Correct rewarded go-trials were associated with widespread activation of midline and lateral barrel cortex areas following a vibration cue and delayed frontal and lateral motor cortex activation. Cortical GCaMP signals predicted trial success and correlated strongly with trial-outcome dependent body movements.

Baropodometric quantification and implications of muscle coactivation in the lower limbs caused by head movement: A prospective study.

BACKGROUND: In healthy young adults, muscle coactivation can sometimes be induced by remote voluntary contractions when the motor task is forceful, maximal, tiring, or cyclic and brief. OBJECTIVES: To show that a change in plantar pressure is an unequivocal response to backward movement of the head, and to contribute to a better understanding of physiotherapy methods that involve remote muscle activation. METHODS: Involuntary coactivation was quantified as a percentage of the anteroposterior plantar pressure distribution, using a baropodometric platform in a population of young adults. The baropodometric data were collected from a 1s recording after 30 s in the reference condition, and from 1s recordings during the first second and then during the 120th second in the test condition. The results were analyzed with Bayesian statistics (Markov chains and Monte Carlo integration techniques). RESULTS: 90 adults participated in the study (age range: 19-26; 38 males and 52 females). The forefoot plantar pressure increased in all cases, by a mean multiplicative factor (on a logit scale) of 1.12 (from 72.24% to 74.45%) when the head was aligned over the trunk. CONCLUSIONS: This 90-participant trial confirmed our initial hypothesis: a increase in forefoot plantar pressure is a systematic response to the motor task (head movement), and suggests greater recruitment of the plantar flexor muscles. A spinal reflex and/or a previously unknown form of motor overflow might be involved in this phenomenon. These results support the development of inductive physiotherapy techniques based on remote muscle activation in the treatment of musculoskeletal disorders. CLINICALSTRIAL. GOV IDENTIFIER: NCT02320097.

Biomechanical Analysis of the Head Movements of a Person Thrown by the Judo Technique 'Seoi-nage'.

The present study examined the kinematics and biomechanical parameters of the head of a person thrown forward by the judo technique 'Seoi-nage'. A judo expert threw an anthropomorphic test device (the POLAR dummy) five times. Kinematics data were obtained with a high-speed digital video camera. Linear and angular accelerations of the head were measured by accelerometers mounted at the center of gravity of the dummy's head. When Seoi-nage was performed, the dummy fell forward accompanied by contacting the anterior parietal regions of the head to the tatami, and the linear and angular accelerations of most axes reached peak values when the head contacted the tatami. Peak resultant linear and angular accelerations were 20.3 ± 9.8 G and 1890.1 ± 1151.9 rad/s2, respectively (means ± standard deviation). Peak values in linear and angular acceleration did not significantly differ between the three directional axes. Absolute angular accelerations in all axes observed in Seoi-nage were high and the resultant value was approximately equal to the already reported in Ouchi-gari, one of the predominant techniques causing judo-related acute subdural hematoma. However, the remarkable increase of linear acceleration in the longitudinal direction and/or angular acceleration in the sagittal plane, as previously reported in techniques being thrown backward (i.e., Ouchi-gari and Osoto-gari), was not detected. The likely mechanism of acute subdural hematoma caused by Seoi-nage is that a large angular acceleration causes large strains and deformations of the brain surface and subsequent rupture of cortical vessels.

Fully automated head-twitch detection system for the study of 5-HT2A receptor pharmacology in vivo.

Head-twitch behavior (HTR) is the behavioral signature of psychedelic drugs upon stimulation of the serotonin 5-HT2A receptor (5-HT2AR) in rodents. Following the previous report of a semi-automated detection of HTR based on the dynamics of mouse's head movement, here we present a system for the identification of individual HTR events in a fully automated fashion. The validity of this fully automated HTR detection system was tested with the psychedelic drug DOI in 5-HT2AR-KO mice, and via evaluation of potential sources of false-positive and false-negative HTR events. The increased throughput in data processing achieved via automation afforded the possibility of conducting otherwise time consuming HTR time-course studies. To further assess the versatility of our system, we also explored the pharmacological interactions between 5-HT2AR and the metabotropic glutamate receptor 2 (mGluR2). Our data demonstrate the potentiation effect of the mGluR2/3 antagonist LY341495 on DOI-induced HTR, as well as the HTR-blocking effect of the mGluR2/3 agonist and antipsychotic drug in development LY404039. This fully automated system can contribute to speed up our understanding of 5-HT2AR's pharmacology and its characteristic behavioral outputs in rodents.

Active head motion reduction in magnetic resonance imaging using tactile feedback.

Head motion is a common problem in clinical as well as empirical (functional) magnetic resonance imaging applications, as it can lead to severe artefacts that reduce image quality. The scanned individuals themselves, however, are often not aware of their head motion. The current study explored whether providing subjects with this information using tactile feedback would reduce their head motion and consequently improve image quality. In a single session that included six runs, 24 participants performed three different cognitive tasks: (a) passive viewing, (b) mental imagery, and (c) speeded responses. These tasks occurred in two different conditions: (a) with a strip of medical tape applied from one side of the magnetic resonance head coil, via the participant's forehead, to the other side, and (b) without the medical tape being applied. Results revealed that application of medical tape to the forehead of subjects to provide tactile feedback significantly reduced both translational as well as rotational head motion. While this effect did not differ between the three cognitive tasks, there was a negative quadratic relationship between head motion with and without feedback. That is, the more head motion a subject produced without feedback, the stronger the motion reduction given the feedback. In conclusion, the here tested method provides a simple and cost-efficient way to reduce subjects' head motion, and might be especially beneficial when extensive head motion is expected a priori.

Is Electrical Nerve Stimulation the Answer for Management of Equine Headshaking?

Horses with trigeminal mediated headshaking (TMHS) have a decreased activation threshold of the trigeminal nerve and clinical signs are suspected to be a manifestation of trigeminal neuralgia. Electrical nerve stimulation (ENS) is used for management of neuralgia in humans and appears to work via gate control theory. Use of an equine specific percutaneous ENS program in over 130 TMHS horses has resulted in approximately 50% success return to previous work. Electroacupuncture may also be useful in the management TMHS. Optimization of ENS procedures for TMHS is likely to require a greater understanding of the etiopathogenesis of the aberrant neurophysiology.

Hallucinogen-Like Action of the Novel Designer Drug 25I-NBOMe and Its Effect on Cortical Neurotransmitters in Rats.

NBOMes are N-benzylmethoxy derivatives of the 2C family hallucinogens. 4-Iodo-2,5-dimethoxy-N-(2-methoxybenzyl)phenethylamine (25I-NBOMe) is one of the commonly used illicit drugs. It exhibits high binding affinity for 5-HT2A/C and 5-HT1A serotonin receptors. Activation of 5-HT2A receptor induces head-twitch response (HTR) in rodents, a behavioral marker of hallucinogen effect in humans. There is not much data on neurochemical properties of NBOMes. Therefore, we aimed to investigate the effect of 25I-NBOMe on extracellular level of dopamine (DA), serotonin (5-HT), and glutamate (GLU) in the rat frontal cortex, tissue contents of monoamines, and hallucinogenic activity in rats. The extracellular levels of DA, 5-HT, and GLU were studied using microdialysis in freely moving animals. The tissue contents of DA, 5-HT and their metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA) were determined in the rat frontal cortex. We also tested a drug-elicited HTR. 25I-NBOMe at doses 1, 3, and 10 mg/kg (sc) increased extracellular DA, 5-HT, and GLU levels, enhanced tissue content of 5-HT and 5-HIAA, but did not affect tissue level of DA and its metabolites. The compound exhibited an inverted U-shaped dose-response curve with respect to the effect on extracellular DA and 5-HT levels, but a U-shaped dose-response curve was observed for its effect on GLU release and HTR. The data from our study suggest that hallucinogenic activity of 25I-NBOMe seems to be related with the increase in extracellular GLU level-mediated via cortical 5-HT2A receptors. The influence of 25I-NBOMe on 5-HT2C and 5-HT1A receptors may modulate its effect on neurotransmitters and HTR.