Functional ultrasound imaging of stroke in awake rats

Anesthesia is a major confounding factor in preclinical stroke research as stroke rarely occurs in sedated patients. Moreover, anesthesia affects both brain functions and the stroke outcome acting as neurotoxic or protective agent. So far, no approaches were well suited to induce stroke while imaging hemodynamics along with simultaneous large-scale recording of brain functions in awake animals. For this reason, the first critical hours following the stroke insult and associated functional alteration remain poorly understood. Here, we present a strategy to investigate both stroke hemodynamics and stroke-induced functional alterations without the confounding effect of anesthesia, i.e., under awake condition. Functional ultrasound (fUS) imaging was used to continuously monitor variations in cerebral blood volume (CBV) in +65 brain regions/hemisphere for up to 3hrs after stroke onset. The focal cortical ischemia was induced using a chemo-thrombotic agent suited for permanent middle cerebral artery occlusion in awake rats, and followed by ipsi- and contralesional whiskers stimulation to investigate on the dynamic of the thalamo-cortical functions. Early (0-3hrs) and delayed (day 5) fUS recording enabled to characterize the features of the ischemia (location, CBV loss), spreading depolarizations (occurrence, amplitude) and functional alteration of the somatosensory thalamo-cortical circuits. Post-stroke thalamo-cortical functions were affected not only early after the stroke onset but were also altered secondarly and remotely from the initial insult. Overall, our procedure enables early, continuous, and chronic evaluations of hemodynamics and brain functions which, combined to stroke or other pathologies, aims to better understand physiopathologies toward the development of clinically relevant therapeutic strategies.

[1]  K. Muir Treatment of wake-up stroke: stick or TWIST? , 2022, Lancet Neurology.

[2]  G. Montaldo,et al.  Functional Ultrasound Neuroimaging. , 2022, Annual review of neuroscience.

[3]  G. Montaldo,et al.  Quantitative Hemodynamic Measurements in Cortical Vessels Using Functional Ultrasound Imaging , 2022, Frontiers in Neuroscience.

[4]  B. Weber,et al.  Vascular Response to Spreading Depolarization Predicts Stroke Outcome , 2022, Stroke.

[5]  M. Endres,et al.  Brain-wide continuous functional ultrasound imaging for real-time monitoring of hemodynamics during ischemic stroke , 2022, bioRxiv.

[6]  M. Gutiérrez-Fernández,et al.  B-Mode Ultrasound, a Reliable Tool for Monitoring Experimental Intracerebral Hemorrhage , 2021, Frontiers in Neurology.

[7]  B. Roska,et al.  Whole-brain functional ultrasound imaging in awake head-fixed mice , 2021, Nature Protocols.

[8]  G. Montaldo,et al.  Optogenetic fUSI for brain-wide mapping of neural activity mediating collicular-dependent behaviors , 2021, Neuron.

[9]  M. Tanter,et al.  Adaptive modulation of brain hemodynamics across stereotyped running episodes , 2020, Nature Communications.

[10]  Clément Brunner,et al.  A Platform for Brain-wide Volumetric Functional Ultrasound Imaging and Analysis of Circuit Dynamics in Awake Mice , 2020, Neuron.

[11]  Ulrich Dirnagl,et al.  The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research* , 2020, BMC Veterinary Research.

[12]  Georgios P. Foustoukos,et al.  Anatomically and functionally distinct thalamocortical inputs to primary and secondary mouse whisker somatosensory cortices , 2020, Nature Communications.

[13]  Georgios P. Foustoukos,et al.  Anatomically and functionally distinct thalamocortical inputs to primary and secondary mouse whisker somatosensory cortices , 2020, Nature Communications.

[14]  Esther J Pearl,et al.  The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research , 2020, PLoS biology.

[15]  M. Tanter,et al.  Early Ultrafast Ultrasound Imaging of Cerebral Perfusion correlates with Ischemic Stroke outcomes and responses to treatment in Mice , 2020, Theranostics.

[16]  P. Drew,et al.  Transfer functions linking neural calcium to single voxel functional ultrasound signal , 2020, Nature Communications.

[17]  Thoralf Niendorf,et al.  The (Un)Conscious Mouse as a Model for Human Brain Functions: Key Principles of Anesthesia and Their Impact on Translational Neuroimaging , 2020, Frontiers in Systems Neuroscience.

[18]  Johannes C. Dahmen,et al.  Subcortical circuits mediate communication between primary sensory cortical areas in mice , 2020, Nature Communications.

[19]  Michelle Y. Cheng,et al.  Inflammatory Responses in the Secondary Thalamic Injury After Cortical Ischemic Stroke , 2020, Frontiers in Neurology.

[20]  A. Hess,et al.  Management. , 2020, Anesthesiology.

[21]  David A Boas,et al.  Awake chronic mouse model of targeted pial vessel occlusion via photothrombosis , 2020, Neurophotonics.

[22]  T. Arumugam,et al.  Motor deficit in the mouse ferric chloride-induced distal middle cerebral artery occlusion model of stroke , 2019, Behavioural Brain Research.

[23]  Mehdi Adibi,et al.  Whisker-Mediated Touch System in Rodents: From Neuron to Behavior , 2019, Front. Syst. Neurosci..

[24]  M. Tanter,et al.  4D functional ultrasound imaging of whole-brain activity in rodents , 2019, Nature Methods.

[25]  Charlie Demené,et al.  Ultrafast Doppler for neonatal brain imaging , 2019, NeuroImage.

[26]  Alan Urban,et al.  Whole-Brain Functional Ultrasound Imaging Reveals Brain Modules for Visuomotor Integration , 2018, Neuron.

[27]  J. Kirsch,et al.  Effects of anesthesia on cerebral blood flow, metabolism, and neuroprotection , 2018, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[28]  J. Baron,et al.  Evidence from functional ultrasound imaging of enhanced contralesional microvascular response to somatosensory stimulation in acute middle cerebral artery occlusion/reperfusion in rats: A marker of ultra-early network reorganization? , 2018, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[29]  Timothy H. Murphy,et al.  Targeted ischemic stroke induction and mesoscopic imaging assessment of blood flow and ischemic depolarization in awake mice , 2017, Neurophotonics.

[30]  Kelly A. Tennant,et al.  Optogenetic rewiring of thalamocortical circuits to restore function in the stroke injured brain , 2017, Nature Communications.

[31]  C. Sommer Ischemic stroke: experimental models and reality , 2017, Acta Neuropathologica.

[32]  J. Baron,et al.  Mapping the dynamics of brain perfusion using functional ultrasound in a rat model of transient middle cerebral artery occlusion , 2017, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[33]  C. Ayata,et al.  Anesthesia in Experimental Stroke Research , 2016, Translational Stroke Research.

[34]  K. C. Brennan,et al.  Susceptibility of Primary Sensory Cortex to Spreading Depolarizations , 2016, The Journal of Neuroscience.

[35]  Olli Gröhn,et al.  Comparison of seven different anesthesia protocols for nicotine pharmacologic magnetic resonance imaging in rat , 2016, European Neuropsychopharmacology.

[36]  Elodie Tiran,et al.  EEG and functional ultrasound imaging in mobile rats , 2015, Nature Methods.

[37]  G. Montaldo,et al.  Real-time imaging of brain activity in freely moving rats using functional ultrasound , 2015, Nature Methods.

[38]  C. Kleinschnitz,et al.  Animal models of ischemic stroke and their application in clinical research , 2015, Drug design, development and therapy.

[39]  Martin Lauritzen,et al.  Spreading Depression, Spreading Depolarizations, and the Cerebral Vasculature. , 2015, Physiological reviews.

[40]  Jean Rossier,et al.  Chronic assessment of cerebral hemodynamics during rat forepaw electrical stimulation using functional ultrasound imaging , 2014, NeuroImage.

[41]  Gábor Kozák,et al.  Imaging Reveals the Focal Area of Spreading Depolarizations and a Variety of Hemodynamic Responses in a Rat Microembolic Stroke Model , 2014, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[42]  G. Tononi,et al.  Diaschisis: past, present, future. , 2014, Brain : a journal of neurology.

[43]  Shanbao Tong,et al.  Induction and imaging of photothrombotic stroke in conscious and freely moving rats , 2014, Journal of biomedical optics.

[44]  Stephanie Taylor,et al.  Induction of ischemic stroke in awake freely moving mice reveals that isoflurane anesthesia can mask the benefits of a neuroprotection therapy , 2014, Front. Neuroenergetics.

[45]  Trygve B. Leergaard,et al.  Brain-wide map of efferent projections from rat barrel cortex , 2014, Front. Neuroinform..

[46]  I. Kanno,et al.  Anesthesia and the Quantitative Evaluation of Neurovascular Coupling , 2012, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[47]  Jian Zhang,et al.  Secondary neurodegeneration in remote regions after focal cerebral infarction: a new target for stroke management? , 2012, Stroke.

[48]  Ofer Levi,et al.  Rapid monitoring of cerebral ischemia dynamics using laser-based optical imaging of blood oxygenation and flow , 2012, Biomedical optics express.

[49]  A. Dunn Laser Speckle Contrast Imaging of Cerebral Blood Flow , 2012, Annals of Biomedical Engineering.

[50]  S. Sherman,et al.  Properties of the thalamic projection from the posterior medial nucleus to primary and secondary somatosensory cortices in the mouse , 2011, Proceedings of the National Academy of Sciences.

[51]  Cullen B. Owens,et al.  Anatomical Pathways Involved in Generating and Sensing Rhythmic Whisker Movements , 2011, Front. Integr. Neurosci..

[52]  I. Macrae,et al.  Preclinical stroke research – advantages and disadvantages of the most common rodent models of focal ischaemia , 2011, British journal of pharmacology.

[53]  W. Pulsinelli,et al.  Metabolic and Perfusion Responses to Recurrent Peri-Infarct Depolarization during Focal Ischemia in the Spontaneously Hypertensive Rat: Dominant Contribution of Sporadic CBF Decrements to Infarct Expansion , 2011, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[54]  M. Fink,et al.  Functional ultrasound imaging of the brain , 2011, Nature Methods.

[55]  Majid H. Mohajerani,et al.  Targeted mini-strokes produce changes in interhemispheric sensory signal processing that are indicative of disinhibition within minutes , 2011, Proceedings of the National Academy of Sciences.

[56]  R. Hornung,et al.  Population-based study of wake-up strokes , 2011, Neurology.

[57]  A. Dunn,et al.  Thrombotic distal middle cerebral artery occlusion produced by topical FeCl3 application: A novel model suitable for intravital microscopy and thrombolysis studies , 2011, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[58]  Afonso C. Silva,et al.  Spatiotemporal Evolution of the Functional Magnetic Resonance Imaging Response to Ultrashort Stimuli , 2011, The Journal of Neuroscience.

[59]  Ferenc Bari,et al.  Multi-modal imaging of anoxic depolarization and hemodynamic changes induced by cardiac arrest in the rat cerebral cortex , 2010, NeuroImage.

[60]  A. Dunn,et al.  Spreading depolarizations cycle around and enlarge focal ischaemic brain lesions , 2010, Brain : a journal of neurology.

[61]  Marc Fisher,et al.  Update of the Stroke Therapy Academic Industry Roundtable Preclinical Recommendations , 2009, Stroke.

[62]  D. Rector,et al.  Conditioned lick behavior and evoked responses using whisker twitches in head restrained rats , 2009, Behavioural Brain Research.

[63]  R. Frostig,et al.  Large-Scale Organization of Rat Sensorimotor Cortex Based on a Motif of Large Activation Spreads , 2008, The Journal of Neuroscience.

[64]  Rosalind Pratt,et al.  Direct, Live Imaging of Cortical Spreading Depression and Anoxic Depolarisation Using a Fluorescent, Voltage-Sensitive Dye , 2008, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[65]  Mathias Hoehn,et al.  Early Prediction of Functional Recovery after Experimental Stroke: Functional Magnetic Resonance Imaging, Electrophysiology, and Behavioral Testing in Rats , 2008, The Journal of Neuroscience.

[66]  B. Connors,et al.  VPM and PoM nuclei of the rat somatosensory thalamus: intrinsic neuronal properties and corticothalamic feedback. , 2007, Cerebral cortex.

[67]  Marc Fisher,et al.  Long-Term Changes of Functional MRI–Based Brain Function, Behavioral Status, and Histopathology After Transient Focal Cerebral Ischemia in Rats , 2006, Stroke.

[68]  John E. W. Mayhew,et al.  Investigating neural–hemodynamic coupling and the hemodynamic response function in the awake rat , 2006, NeuroImage.

[69]  F. Ebner,et al.  Chronic suppression of activity in barrel field cortex downregulates sensory responses in contralateral barrel field cortex. , 2005, Journal of neurophysiology.

[70]  M. Moskowitz,et al.  Laser Speckle Flowmetry for the Study of Cerebrovascular Physiology in Normal and Ischemic Mouse Cortex , 2004, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[71]  Simona Temereanca,et al.  Functional Topography of Corticothalamic Feedback Enhances Thalamic Spatial Response Tuning in the Somatosensory Whisker/Barrel System , 2004, Neuron.

[72]  F. Ebner,et al.  Lesions of Mature Barrel Field Cortex Interfere with Sensory Processing and Plasticity in Connected Areas of the Contralateral Hemisphere , 2003, The Journal of Neuroscience.

[73]  Rick M Dijkhuizen,et al.  Correlation between Brain Reorganization, Ischemic Damage, and Neurologic Status after Transient Focal Cerebral Ischemia in Rats: A Functional Magnetic Resonance Imaging Study , 2003, The Journal of Neuroscience.

[74]  Chris J. Martin,et al.  Optical imaging spectroscopy in the unanaesthetised rat , 2002, Journal of Neuroscience Methods.

[75]  B R Rosen,et al.  Functional magnetic resonance imaging of reorganization in rat brain after stroke , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[76]  T. Reese,et al.  Functional recovery after brain lesion—contralateral neuromodulation: an fMRI study , 2001, Neuroreport.

[77]  M. Deschenes,et al.  Corticothalamic Projections from the Cortical Barrel Field to the Somatosensory Thalamus in Rats: A Single‐fibre Study Using Biocytin as an Anterograde Tracer , 1995, The European journal of neuroscience.

[78]  Toshiharu Asai,et al.  Functional Changes in Thalamic Relay Neurons after Focal Cerebral Infarct: A Study of Unit Recordings from VPL Neurons after MCA Occlusion in Rats , 1992, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[79]  Mara Fabri,et al.  Ipsilateral cortical connections of primary somatic sensory cortex in rats , 1991, The Journal of comparative neurology.

[80]  H. Seers,et al.  OUTCOME , 1977, How to Win Your Case.

[81]  M. Carandini,et al.  Neural basis of functional ultrasound signals , 2021 .

[82]  R. Traystman Effect of Anesthesia in Stroke Models , 2010 .

[83]  Colin Camerer : Past , Present , Future , 2003 .

[84]  G. Paxinos,et al.  The Rat Brain in Stereotaxic Coordinates , 1983 .