Neuronavigated Repetitive Transcranial Ultrasound Stimulation Induces Long-Lasting and Reversible Effects on Oculomotor Performance in Non-human Primates.

Since the late 2010s, Transcranial Ultrasound Stimulation (TUS) has been used experimentally to carryout safe, non-invasive stimulation of the brain with better spatial resolution than Transcranial Magnetic Stimulation (TMS). This innovative stimulation method has emerged as a novel and valuable device for studying brain function in humans and animals. In particular, single pulses of TUS directed to oculomotor regions have been shown to modulate visuomotor behavior of non-human primates during 100 ms ultrasound pulses. In the present study, a sustained effect was induced by applying 20-s trains of neuronavigated repetitive Transcranial Ultrasound Stimulation (rTUS) to oculomotor regions of the frontal cortex in three non-human primates performing an antisaccade task. With the help of MRI imaging and a frame-less stereotactic neuronavigation system (SNS), we were able to demonstrate that neuronavigated TUS (outside of the MRI scanner) is an efficient tool to carry out neuromodulation procedures in non-human primates. We found that, following neuronavigated rTUS, saccades were significantly modified, resulting in shorter latencies compared to no-rTUS trials. This behavioral modulation was maintained for up to 20 min. Oculomotor behavior returned to baseline after 18-31 min and could not be significantly distinguished from the no-rTUS condition. This study is the first to show that neuronavigated rTUS can have a persistent effect on monkey behavior with a quantified return-time to baseline. The specificity of the effects could not be explained by auditory confounds.

Non-invasive ultrasonic modulation of visual evoked response by GABA delivery through the blood brain barrier.

GABA is an inhibitory neurotransmitter that is maintained outside the brain by the blood brain barrier in normal condition. In this paper we demonstrate the feasibility of modulating brain activity in the visual cortex of non-human primates by transiently permeabilizing the blood brain barrier (BBB) using focused ultrasound (FUS) coupled with ultrasound contrast agents (UCA), followed by intra-venous injection of GABA. The visual evoked potentials exhibited a significant and GABA-dose-depend decrease in activity. The effect of the sonication only (with and without UCA) was also investigated and was shown to decrease the activity 8.7 times less than the GABA-induced inhibition enabled by BBB permeabilization. Finally, the UCA harmonic response was monitored during sonication to estimate the level of stable cavitation (a signature of the effectiveness of BBB permeabilization) and to avoid damage due to inertial cavitation (the sonication was automatically shut down when this condition was detected). Our results extend the promise of the exploration and treatment of the brain using non-invasive, controllable, repeatable, and reversible neuromodulation.

MRI monitoring of temperature and displacement for transcranial focus ultrasound applications.

BACKGROUND: Transcranial focus ultrasound applications applied under MRI-guidance benefit from unrivaled monitoring capabilities, allowing the recording of real-time anatomical information and biomarkers like the temperature rise and/or displacement induced by the acoustic radiation force. Having both of these measurements could allow for better targeting of brain structures, with improved therapy monitoring and safety. METHOD: We investigated the use of a novel MRI-pulse sequence described previously in Bour et al., (2017) to quantify both the displacement and temperature changes under various ultrasound sonication conditions and in different regions of the brain. The method was evaluated in vivo in a non-human primate under anesthesia using a single-element transducer (f = 850 kHz) in a setting that could mimic clinical applications. Acquisition was performed at 3 T on a clinical imaging system using a modified single-shot gradient echo EPI sequence integrating a bipolar motion-sensitive encoding gradient. Four slices were acquired sequentially perpendicularly or axially to the direction of the ultrasound beam with a 1-Hz update frequency and an isotropic spatial resolution of 2-mm. A total of twenty-four acquisitions were performed in three different sets of experiments. Measurement uncertainty of the sequence was investigated under different acoustic power deposition and in different regions of the brain. Acoustic simulation and thermal modeling were performed and compared to experimental data. RESULTS: The sequence simultaneously provides relevant information about the focal spot location and visualization of heating of brain structures: 1) The sequence localized the acoustic focus both along as well as perpendicular to the ultrasound direction. Tissue displacements ranged from 1 to 2 µm. 2) Thermal rise was only observed at the vicinity of the skull. Temperature increase ranged between 1 and 2 °C and was observed delayed relative the sonication due to thermal diffusion. 3) The fast frame rate imaging was able to highlight magnetic susceptibility artifacts related to breathing, for the most caudal slices. We demonstrated that respiratory triggering successfully restored the sensitivity of the method (from 0.7 µm to 0.2 µm). 4) These results were corroborated by acoustic simulations. CONCLUSIONS: The current rapid, multi-slice acquisition and real-time implementation of temperature and displacement visualization may be useful in clinical practices. It may help defining operational safety margins, improving therapy precision and efficacy. Simulations were in good agreement with experimental data and may thus be used prior treatment for procedure planning.

Offline impact of transcranial focused ultrasound on cortical activation in primates.

To understand brain circuits it is necessary both to record and manipulate their activity. Transcranial ultrasound stimulation (TUS) is a promising non-invasive brain stimulation technique. To date, investigations report short-lived neuromodulatory effects, but to deliver on its full potential for research and therapy, ultrasound protocols are required that induce longer-lasting 'offline' changes. Here, we present a TUS protocol that modulates brain activation in macaques for more than one hour after 40 s of stimulation, while circumventing auditory confounds. Normally activity in brain areas reflects activity in interconnected regions but TUS caused stimulated areas to interact more selectively with the rest of the brain. In a within-subject design, we observe regionally specific TUS effects for two medial frontal brain regions - supplementary motor area and frontal polar cortex. Independently of these site-specific effects, TUS also induced signal changes in the meningeal compartment. TUS effects were temporary and not associated with microstructural changes.

Manipulation of Subcortical and Deep Cortical Activity in the Primate Brain Using Transcranial Focused Ultrasound Stimulation.

The causal role of an area within a neural network can be determined by interfering with its activity and measuring the impact. Many current reversible manipulation techniques have limitations preventing their application, particularly in deep areas of the primate brain. Here, we demonstrate that a focused transcranial ultrasound stimulation (TUS) protocol impacts activity even in deep brain areas: a subcortical brain structure, the amygdala (experiment 1), and a deep cortical region, the anterior cingulate cortex (ACC, experiment 2), in macaques. TUS neuromodulatory effects were measured by examining relationships between activity in each area and the rest of the brain using functional magnetic resonance imaging (fMRI). In control conditions without sonication, activity in a given area is related to activity in interconnected regions, but such relationships are reduced after sonication, specifically for the targeted areas. Dissociable and focal effects on neural activity could not be explained by auditory confounds.

Potential impact of thermal effects during ultrasonic neurostimulation: retrospective numerical estimation of temperature elevation in seven rodent setups.

In the past decade, a handful but growing number of groups have reported worldwide successful low intensity focused ultrasound induced neurostimulation trials on rodents. Its effects range from movement elicitations to reduction of anesthesia time or reduction of the duration of drug induced seizures. The mechanisms underlying ultrasonic neuromodulation are still not fully understood. Given the low intensities used in most of the studies, a mechanical effect is more likely to be responsible for the neuromodulation effect, but a clear description of the thermal and mechanical effects is necessary to optimize clinical applications. Based on five studies settings, we calculated the temperature rise and thermal doses in order to evaluate its implication in the neuromodulation phenomenon. Our retrospective analysis shows thermal rise ranging from 0.002 °C to 0.8 °C in the brain for all setups, except for one setup for which the temperature increase is estimated to be as high as 7 °C. We estimate that in the latter case, temperature rise cannot be neglected as a possible cause of neuromodulation. Simulations results were supported by temperature measurements on a mouse with two different sets of parameters. Although the calculated temperature is compatible with the absence of visible thermal lesions on the skin, it is high enough to impact brain circuits. Our study highlights the usefulness of performing thermal simulations prior to experiment in order to fully take into account not only the impact of the peak intensity but also pulse duration and pulse repetition.

Transcranial ultrasonic stimulation modulates single-neuron discharge in macaques performing an antisaccade task.

BACKGROUND: Low intensity transcranial ultrasonic stimulation (TUS) has been demonstrated to non-invasively and transiently stimulate the nervous system. Although US neuromodulation has appeared robust in rodent studies, the effects of US in large mammals and humans have been modest at best. In addition, there is a lack of direct recordings from the stimulated neurons in response to US. Our study investigates the magnitude of the US effects on neuronal discharge in awake behaving monkeys and thus fills the void on both fronts. OBJECTIVE/HYPOTHESIS: In this study, we demonstrate the feasibility of recording action potentials in the supplementary eye field (SEF) as TUS is applied simultaneously to the frontal eye field (FEF) in macaques performing an antisaccade task. RESULTS: We show that compared to a control stimulation in the visual cortex, SEF activity is significantly modulated shortly after TUS onset. Among all cell types 40% of neurons significantly changed their activity after TUS. Half of the neurons showed a transient increase of activity induced by TUS. CONCLUSION: Our study demonstrates that the neuromodulatory effects of non-invasive focused ultrasound can be assessed in real time in awake behaving monkeys by recording discharge activity from a brain region reciprocally connected with the stimulated region. The study opens the door for further parametric studies for fine-tuning the ultrasonic parameters. The ultrasonic effect could indeed be quantified based on the direct measurement of the intensity of the modulation induced on a single neuron in a freely performing animal. The technique should be readily reproducible in other primate laboratories studying brain function, both for exploratory and therapeutic purposes and to facilitate the development of future clinical TUS devices.

Erratum to "A 200-1380 kHz Quadrifrequency Focused Ultrasound Transducer for Neurostimulation in Rodents and Primates: Transcranial In Vitro Calibration and Numerical Study of the Influence of Skull Cavity".

In the above paper [1], one maximum pressure listed in Table I, page 719, should be corrected. This error occurred when reporting the maximum pressure estimated in the rat brain at 1380 kHz (line 4, last column). The right value (7 MPa) does not change the discussion, and is in line with the 83% estimated pressure gain that was initially reported in rat brain (line 7, last column). Here we provide the correct table.

A 200-1380-kHz Quadrifrequency Focused Ultrasound Transducer for Neurostimulation in Rodents and Primates: Transcranial In Vitro Calibration and Numerical Study of the Influence of Skull Cavity.

Low intensity transcranial focused ultrasound has been demonstrated to produce neuromodulation in both animals and humans. Primarily for technical reasons, frequency is one of the most poorly investigated critical wave parameters. We propose the use of a quadri-band transducer capable of operating at 200, 320, 850, and 1380 kHz for further investigation of the frequency dependence of neuromodulation efficacy while keeping the position of the transducer fixed with respect to the subject's head. This paper presents the results of the transducer calibration in water, in vitro transmission measurements through a monkey skull flap, 3-D simulations based on both a µ -computed tomography ( µ CT)-scan of a rat and on CT-scans of two macaques. A maximum peak pressure greater than 0.52 MPa is expected at each frequency in rat and macaque heads. According to the literature, our transducer can achieve neuromodulation in rodents and primates at each four frequencies. The impact of standing waves is shown to be most prominent at the lowest frequencies.

Scalable Graphene-Based Membranes for Ionic Sieving with Ultrahigh Charge Selectivity.

Nanostructured graphene-oxide (GO) laminate membranes, exhibiting ultrahigh water flux, are excellent candidates for next generation nanofiltration and desalination membranes, provided the ionic rejection could be further increased without compromising the water flux. Using microscopic drift-diffusion experiments, we demonstrated the ultrahigh charge selectivity for GO membranes, with more than order of magnitude difference in the permeabilities of cationic and anionic species of equivalent hydration radii. Measuring diffusion of a wide range of ions of different size and charge, we were able to clearly disentangle different physical mechanisms contributing to the ionic sieving in GO membranes: electrostatic repulsion between ions and charged chemical groups; and the compression of the ionic hydration shell within the membrane's nanochannels, following the activated behavior. The charge-selectivity allows us to rationally design membranes with increased ionic rejection and opens up the field of ion exchange and electrodialysis to the GO membranes.