{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T17:09:40Z","timestamp":1781024980673,"version":"3.54.1"},"reference-count":52,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T00:00:00Z","timestamp":1670889600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000104","name":"National Aeronautics and Space Administration","doi-asserted-by":"publisher","award":["NNX16AO69A-T0415"],"award-info":[{"award-number":["NNX16AO69A-T0415"]}],"id":[{"id":"10.13039\/100000104","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Low-intensity transcranial focused ultrasound (tFUS) offers new functional neuromodulation opportunities, enabling stimulation of cortical as well as deep brain areas with high spatial resolution. Brain stimulation of awake sheep, in the absence of the confounding effects of anesthesia on brain function, provides translational insight into potential human applications with safety information supplemented by histological analyses. We examined the effects of tFUS pulsing parameters, particularly regarding pulse durations (PDs), on stimulating the cortical motor area (M1) and its thalamic projection in unanesthetized, awake sheep (<jats:italic>n<\/jats:italic> = 8). A wearable tFUS headgear, custom-made for individual sheep, enabled experiments to be conducted without using anesthesia. FUS stimuli, each 200 ms long, were delivered to the M1 and the thalamus using three different PDs (0.5, 1, and 2 ms) with the pulse repetition frequency (PRF) adjusted to maintain a 70% duty cycle at a derated <jats:italic>in situ<\/jats:italic> spatial-peak temporal-average intensity (I<jats:sub>spta<\/jats:sub>) of 3.6 W\/cm<jats:sup>2<\/jats:sup>. Efferent electromyography (EMG) responses to stimulation were quantified from both hind limbs. Group-averaged EMG responses from each of the hind limbs across the experimental conditions revealed selective responses from the hind limb contralateral to sonication. The use of 0.5 and 1 ms PDs generated higher EMG signal amplitudes compared to those obtained using a 2 ms PD. Faster efferent response was also observed from thalamic stimulation than that from stimulating the M1. Post-sonication behavioral observation and histological assessment performed 24 h and 1 month after sonication were not indicative of any abnormalities. The results suggest the presence of pulsing scheme-dependent effects of tFUS on brain stimulation and attest its safety in awake large animals.<\/jats:p>","DOI":"10.1371\/journal.pone.0278865","type":"journal-article","created":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T18:58:08Z","timestamp":1670957888000},"page":"e0278865","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":11,"title":["Effects of focused ultrasound pulse duration on stimulating cortical and subcortical motor circuits in awake sheep"],"prefix":"10.1371","volume":"17","author":[{"given":"Hyun-Chul","family":"Kim","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wonhye","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kavin","family":"Kowsari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel S.","family":"Weisholtz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5150-9857","authenticated-orcid":true,"given":"Seung-Schik","family":"Yoo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"issue":"7","key":"pone.0278865.ref001","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1016\/j.ultrasmedbio.2018.12.015","article-title":"Ultrasound neuromodulation: a review of results, mechanisms and safety","volume":"45","author":"J Blackmore","year":"2019","journal-title":"Ultrasound Med Biol"},{"issue":"2","key":"pone.0278865.ref002","doi-asserted-by":"crossref","first-page":"e16","DOI":"10.12786\/bn.2018.11.e16","article-title":"Technical review and perspectives of transcranial focused ultrasound brain stimulation for neurorehabilitation","volume":"11","author":"SS Yoo","year":"2018","journal-title":"Brain Neurorehabil"},{"issue":"3","key":"pone.0278865.ref003","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.brs.2011.03.007","article-title":"A review of low-intensity focused ultrasound pulsation","volume":"4","author":"A Bystritsky","year":"2011","journal-title":"Brain Stimul"},{"issue":"15","key":"pone.0278865.ref004","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1097\/WNR.0b013e32834b2957","article-title":"Transcranial focused ultrasound to the thalamus alters anesthesia time in rats","volume":"22","author":"S-S Yoo","year":"2011","journal-title":"Neuroreport"},{"issue":"2","key":"pone.0278865.ref005","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.ultrasmedbio.2012.09.009","article-title":"Effective parameters for ultrasound-induced in vivo neurostimulation","volume":"39","author":"RL King","year":"2013","journal-title":"Ultrasound Med Biol"},{"issue":"10","key":"pone.0278865.ref006","doi-asserted-by":"crossref","first-page":"5730","DOI":"10.1118\/1.4963208","article-title":"Focused ultrasound neuromodulation of cortical and subcortical brain structures using 1.9 MHz","volume":"43","author":"HA Kamimura","year":"2016","journal-title":"Med Phys"},{"issue":"5","key":"pone.0278865.ref007","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/j.brs.2014.06.011","article-title":"Focused ultrasound-mediated non-invasive brain stimulation: examination of sonication parameters","volume":"7","author":"H Kim","year":"2014","journal-title":"Brain Stimul"},{"issue":"3","key":"pone.0278865.ref008","doi-asserted-by":"crossref","first-page":"875","DOI":"10.3171\/2016.11.JNS16976","article-title":"Noninvasive neuromodulation and thalamic mapping with low-intensity focused ultrasound","volume":"128","author":"RF Dallapiazza","year":"2017","journal-title":"J Neurosurg"},{"issue":"10","key":"pone.0278865.ref009","doi-asserted-by":"crossref","first-page":"e0224311","DOI":"10.1371\/journal.pone.0224311","article-title":"Effects of sonication parameters on transcranial focused ultrasound brain stimulation in an ovine model","volume":"14","author":"K Yoon","year":"2019","journal-title":"PLoS One"},{"issue":"2","key":"pone.0278865.ref010","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/j.ultrasmedbio.2015.10.001","article-title":"Image-guided focused ultrasound-mediated regional brain stimulation in sheep","volume":"42","author":"W Lee","year":"2016","journal-title":"Ultrasound Med Biol"},{"key":"pone.0278865.ref011","doi-asserted-by":"crossref","first-page":"e40541","DOI":"10.7554\/eLife.40541","article-title":"Offline impact of transcranial focused ultrasound on cortical activation in primates","volume":"8","author":"L Verhagen","year":"2019","journal-title":"Elife"},{"issue":"6","key":"pone.0278865.ref012","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1016\/j.neuron.2019.01.019","article-title":"Manipulation of subcortical and deep cortical activity in the primate brain using transcranial focused ultrasound stimulation","volume":"101","author":"D Folloni","year":"2019","journal-title":"Neuron"},{"key":"pone.0278865.ref013","doi-asserted-by":"crossref","first-page":"8743","DOI":"10.1038\/srep08743","article-title":"Image-guided transcranial focused ultrasound stimulates human primary somatosensory cortex","volume":"5","author":"W Lee","year":"2015","journal-title":"Sci Rep"},{"issue":"2","key":"pone.0278865.ref014","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1038\/nn.3620","article-title":"Transcranial focused ultrasound modulates the activity of primary somatosensory cortex in humans","volume":"17","author":"W Legon","year":"2014","journal-title":"Nat Neurosci"},{"issue":"6","key":"pone.0278865.ref015","doi-asserted-by":"crossref","first-page":"e0178476","DOI":"10.1371\/journal.pone.0178476","article-title":"Non-invasive transmission of sensorimotor information in humans using an EEG\/focused ultrasound brain-to-brain interface","volume":"12","author":"W Lee","year":"2017","journal-title":"PLoS One"},{"issue":"1","key":"pone.0278865.ref016","doi-asserted-by":"crossref","first-page":"34026","DOI":"10.1038\/srep34026","article-title":"Transcranial focused ultrasound stimulation of human primary visual cortex","volume":"6","author":"W Lee","year":"2016","journal-title":"Sci Rep"},{"issue":"1","key":"pone.0278865.ref017","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s12868-018-0456-6","article-title":"Effects of transcranial focused ultrasound on human primary motor cortex using 7T fMRI: a pilot study","volume":"19","author":"L Ai","year":"2018","journal-title":"BMC Neurosci"},{"issue":"6","key":"pone.0278865.ref018","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1016\/j.brs.2016.07.008","article-title":"Non-invasive ultrasonic thalamic stimulation in disorders of consciousness after severe brain injury: A first-in-man report","volume":"9","author":"MM Monti","year":"2016","journal-title":"Brain Stimul"},{"issue":"5","key":"pone.0278865.ref019","doi-asserted-by":"crossref","first-page":"1270","DOI":"10.1016\/j.ultrasmedbio.2020.01.007","article-title":"Focused ultrasound platform for investigating therapeutic neuromodulation across the human hippocampus","volume":"46","author":"ST Brinker","year":"2020","journal-title":"Ultrasound Med Biol"},{"issue":"3","key":"pone.0278865.ref020","doi-asserted-by":"crossref","first-page":"1267","DOI":"10.1016\/j.neuroimage.2011.02.058","article-title":"Focused ultrasound modulates region-specific brain activity","volume":"56","author":"SS Yoo","year":"2011","journal-title":"Neuroimage"},{"key":"pone.0278865.ref021","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1186\/1471-2202-12-23","article-title":"Focused ultrasound-mediated suppression of chemically-induced acute epileptic EEG activity","volume":"12","author":"BK Min","year":"2011","journal-title":"BMC Neurosci"},{"issue":"3","key":"pone.0278865.ref022","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1016\/j.brs.2020.02.017","article-title":"Histologic safety of transcranial focused ultrasound neuromodulation and magnetic resonance acoustic radiation force imaging in rhesus macaques and sheep","volume":"13","author":"P Gaur","year":"2020","journal-title":"Brain Stimul"},{"issue":"1","key":"pone.0278865.ref023","doi-asserted-by":"crossref","first-page":"15347","DOI":"10.1038\/s41598-020-72189-y","article-title":"Ultrasound neuromodulation depends on pulse repetition frequency and can modulate inhibitory effects of TTX","volume":"10","author":"TJ Manuel","year":"2020","journal-title":"Sci Rep"},{"issue":"5","key":"pone.0278865.ref024","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.neuron.2010.05.008","article-title":"Transcranial pulsed ultrasound stimulates intact brain circuits","volume":"66","author":"Y Tufail","year":"2010","journal-title":"Neuron"},{"issue":"1","key":"pone.0278865.ref025","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1186\/s12868-018-0459-3","article-title":"Transcranial focused ultrasound stimulation of motor cortical areas in freely-moving awake rats","volume":"19","author":"W Lee","year":"2018","journal-title":"BMC Neurosci"},{"issue":"10","key":"pone.0278865.ref026","doi-asserted-by":"crossref","first-page":"e3511","DOI":"10.1371\/journal.pone.0003511","article-title":"Remote excitation of neuronal circuits using low-intensity, low-frequency ultrasound","volume":"3","author":"WJ Tyler","year":"2008","journal-title":"PLoS One"},{"issue":"1","key":"pone.0278865.ref027","doi-asserted-by":"crossref","first-page":"19274","DOI":"10.1038\/s41598-021-98920-x","article-title":"Transcranial focused ultrasound modulates cortical and thalamic motor activity in awake sheep","volume":"11","author":"HC Kim","year":"2021","journal-title":"Sci Rep"},{"issue":"1","key":"pone.0278865.ref028","doi-asserted-by":"crossref","first-page":"100760","DOI":"10.1016\/j.isci.2019.100760","article-title":"The general anesthetic isoflurane bilaterally modulates neuronal excitability","volume":"23","author":"M Ou","year":"2020","journal-title":"iScience"},{"issue":"6","key":"pone.0278865.ref029","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1097\/00000542-197306000-00011","article-title":"Neurophysiologic effects of general anesthetics: I. The electroencephalogram and sensory evoked responses in man","volume":"38","author":"DL Clark","year":"1973","journal-title":"Anesthesiology"},{"key":"pone.0278865.ref030","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.jelekin.2015.12.005","article-title":"Surface electromyography in animal biomechanics: A systematic review","volume":"28","author":"S Valentin","year":"2016","journal-title":"J Electromyogr Kinesiol"},{"issue":"10","key":"pone.0278865.ref031","doi-asserted-by":"crossref","first-page":"105001","DOI":"10.1088\/1361-6560\/aabe37","article-title":"Multi-resolution simulation of focused ultrasound propagation through ovine skull from a single-element transducer","volume":"63","author":"K Yoon","year":"2018","journal-title":"Phys Med Biol"},{"issue":"12","key":"pone.0278865.ref032","doi-asserted-by":"crossref","first-page":"3379","DOI":"10.1007\/s00221-018-5370-8","article-title":"Indices of comparative cognition: assessing animal models of human brain function","volume":"236","author":"SD McBride","year":"2018","journal-title":"Exp Brain Res"},{"issue":"1\u20132","key":"pone.0278865.ref033","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0304-3940(94)90589-4","article-title":"Abnormally phosphorylated tau protein related to the formation of neurofibrillary tangles and neuropil threads in the cerebral cortex of sheep and goat","volume":"171","author":"H Braak","year":"1994","journal-title":"Neurosci Lett"},{"key":"pone.0278865.ref034","doi-asserted-by":"crossref","first-page":"587","DOI":"10.3389\/fnins.2019.00587","article-title":"Determining the temporal profile of intracranial pressure changes following transient stroke in an ovine model","volume":"13","author":"AJ Sorby-Adams","year":"2019","journal-title":"Front Neurosci"},{"issue":"3","key":"pone.0278865.ref035","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.brs.2014.02.002","article-title":"Brain stimulation for epilepsy\u2013local and remote modulation of network excitability","volume":"7","author":"PH Stypulkowski","year":"2014","journal-title":"Brain Stimul"},{"issue":"3","key":"pone.0278865.ref036","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1002\/mus.20333","article-title":"Comparing the function of the corticospinal system in different species: organizational differences for motor specialization?","volume":"32","author":"RN Lemon","year":"2005","journal-title":"Muscle Nerve"},{"issue":"5","key":"pone.0278865.ref037","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1016\/j.humov.2010.07.007","article-title":"Using transcranial magnetic stimulation methods to probe connectivity between motor areas of the brain","volume":"30","author":"JC Rothwell","year":"2011","journal-title":"Hum Mov Sci"},{"issue":"4","key":"pone.0278865.ref038","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1007\/s002210050648","article-title":"Direct demonstration of interhemispheric inhibition of the human motor cortex produced by transcranial magnetic stimulation","volume":"124","author":"V Di Lazzaro","year":"1999","journal-title":"Exp Brain Res"},{"issue":"12","key":"pone.0278865.ref039","doi-asserted-by":"crossref","first-page":"1309","DOI":"10.1007\/s11517-012-0965-x","article-title":"A method for determination of muscle fiber diameter using single fiber potential (SFP) analysis.","volume":"50","author":"E Zalewska","year":"2012","journal-title":"Med Biol Eng Comput"},{"issue":"24","key":"pone.0278865.ref040","doi-asserted-by":"crossref","first-page":"10966","DOI":"10.1523\/JNEUROSCI.22-24-10966.2002","article-title":"Feedforward mechanisms of excitatory and inhibitory cortical receptive fields","volume":"22","author":"RM Bruno","year":"2002","journal-title":"J Neurosci"},{"issue":"6","key":"pone.0278865.ref041","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1097\/00004728-200311000-00001","article-title":"Magnetic resonance: an introduction to ultrashort TE (UTE) imaging.","volume":"27","author":"MD Robson","year":"2003","journal-title":"J Comput Assist Tomogr"},{"key":"pone.0278865.ref042","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1007\/978-3-540-74658-4_16","volume-title":"Springer handbook of medical technology:","author":"TM Buzug","year":"2011"},{"issue":"17","key":"pone.0278865.ref043","first-page":"5324","article-title":"Bidirectional regulation of protein kinase M\u03b6 in the maintenance of long-term potentiation and long-term depression","volume":"16","author":"S Hrabetova","year":"1996","journal-title":"J Neurosci Res"},{"issue":"2","key":"pone.0278865.ref044","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.brs.2021.01.006","article-title":"Bidirectional and state-dependent modulation of brain activity by transcranial focused ultrasound in non-human primates","volume":"14","author":"P-F Yang","year":"2021","journal-title":"Brain Stimul"},{"issue":"1","key":"pone.0278865.ref045","doi-asserted-by":"crossref","first-page":"2519","DOI":"10.1038\/s41467-021-22743-7","article-title":"Intrinsic functional neuron-type selectivity of transcranial focused ultrasound neuromodulation.","volume":"12","author":"K Yu","year":"2021","journal-title":"Nat Commun"},{"issue":"2","key":"pone.0278865.ref046","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1016\/j.brs.2022.01.014","article-title":"Long term study of motivational and cognitive effects of low-intensity focused ultrasound neuromodulation in the dorsal striatum of nonhuman primates","volume":"15","author":"F Munoz","year":"2022","journal-title":"Brain Stimul"},{"issue":"6","key":"pone.0278865.ref047","doi-asserted-by":"crossref","first-page":"1527","DOI":"10.1016\/j.brs.2020.08.014","article-title":"Transcranial ultrasound stimulation in humans is associated with an auditory confound that can be effectively masked","volume":"13","author":"V Braun","year":"2020","journal-title":"Brain Stimul"},{"issue":"5","key":"pone.0278865.ref048","doi-asserted-by":"crossref","first-page":"1031","DOI":"10.1016\/j.neuron.2018.05.009","article-title":"Ultrasonic neuromodulation causes widespread cortical activation via an indirect auditory mechanism","volume":"98","author":"T Sato","year":"2018","journal-title":"Neuron"},{"issue":"3","key":"pone.0278865.ref049","doi-asserted-by":"crossref","DOI":"10.1523\/ENEURO.0136-15.2016","article-title":"Cell-type-selective effects of intramembrane cavitation as a unifying theoretical framework for ultrasonic neuromodulation","volume":"3","author":"M Plaksin","year":"2016","journal-title":"Eneuro"},{"issue":"1","key":"pone.0278865.ref050","first-page":"011004","article-title":"Intramembrane cavitation as a predictive bio-piezoelectric mechanism for ultrasonic brain stimulation","volume":"4","author":"M Plaksin","year":"2014","journal-title":"Phys Rev X"},{"issue":"1","key":"pone.0278865.ref051","first-page":"1","article-title":"Focused ultrasound excites cortical neurons via mechanosensitive calcium accumulation and ion channel amplification","volume":"13","author":"S Yoo","year":"2022","journal-title":"Nat Commun"},{"issue":"1","key":"pone.0278865.ref052","first-page":"1","article-title":"Inhibitory effect of ultrasonic stimulation on the voltage-dependent potassium currents in rat hippocampal CA1 neurons","volume":"20","author":"K Cui","year":"2019","journal-title":"BMC Neurosci"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0278865","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T18:58:52Z","timestamp":1670957932000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0278865"}},"subtitle":[],"editor":[{"given":"John A.","family":"Thompson","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2022,12,13]]},"references-count":52,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12,13]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0278865","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,13]]}}}