Multimodal assessment of recovery from coma in a rat model of diffuse brainstem tegmentum injury

ABSTRACT Despite the association between brainstem lesions and coma, a mechanistic understanding of coma pathogenesis and recovery is lacking. We developed a coma model in the rat mimicking human brainstem coma, which allowed multimodal analysis of a brainstem tegmentum lesion's effects on behavior, cortical electrophysiology, and global brain functional connectivity. After coma induction, we observed a transient period (˜1h) of unresponsiveness accompanied by cortical burst‐suppression. Comatose rats then gradually regained behavioral responsiveness concurrent with emergence of delta/theta‐predominant cortical rhythms in primary somatosensory cortex. During the acute stage of coma recovery (˜1–8h), longitudinal resting‐state functional MRI revealed an increase in functional connectivity between subcortical arousal nuclei in the thalamus, basal forebrain, and basal ganglia and cortical regions implicated in awareness. This rat coma model provides an experimental platform to systematically study network‐based mechanisms of coma pathogenesis and recovery, as well as to test targeted therapies aimed at promoting recovery of consciousness after coma. HIGHLIGHTSET‐1‐induced diffuse brainstem injury produced rat coma mimicking human brainstem coma.The Rat Coma Scale and LFP track acute recovery of neurological function in comatose rats.Rat‐MRI characterized the anatomical lesion and functional restoration after coma.The cortex reactivates as subcortical centers regain functional connectivity.

[1]  Ciprian Catana,et al.  Disruption of thalamic functional connectivity is a neural correlate of dexmedetomidine-induced unconsciousness , 2014, eLife.

[2]  Michael M. Halassa,et al.  Selective optical drive of thalamic reticular nucleus generates thalamic bursts & cortical spindles , 2011, Nature Neuroscience.

[3]  W. Bamlet,et al.  FOUR score and Glasgow Coma Scale in predicting outcome of comatose patients: A pooled analysis , 2011, Neurology.

[4]  A. Damasio,et al.  Neuroanatomical correlates of brainstem coma. , 2003, Brain : a journal of neurology.

[5]  Matthew Jeffers,et al.  A reproducible Endothelin-1 model of forelimb motor cortex stroke in the mouse , 2014, Journal of Neuroscience Methods.

[6]  F Robert,et al.  [Criteria of brain death. Spinal reflexes in 45 personal studies]. , 1977, Schweizerische medizinische Wochenschrift.

[7]  Michael Chopp,et al.  Animal models of traumatic brain injury , 2013, Nature Reviews Neuroscience.

[8]  Richard P Brenner,et al.  The Interpretation of the EEG in Stupor and Coma , 2005, The neurologist.

[9]  J. Woods.  BEHAVIOR OF CHRONIC DECEREBRATE RATS. , 1964, Journal of neurophysiology.

[10]  Ezequiel Mikulan,et al.  The anterior cingulate cortex: an integrative hub for human socially-driven interactions , 2013, Front. Neurosci..

[11]  H Gromysz,et al.  The effects of brainstem transection on respiratory activity in the rabbit. , 1981, Acta neurobiologiae experimentalis.

[12]  W. Rosenblum,et al.  Immediate, irreversible, posttraumatic coma: a review indicating that bilateral brainstem injury rather than widespread hemispheric damage is essential for its production. , 2015, Journal of neuropathology and experimental neurology.

[13]  Sadao Kimura,et al.  A novel potent vasoconstrictor peptide produced by vascular endothelial cells , 1988, Nature.

[14]  V. Synek,et al.  Prognostically Important EEG Coma Patterns in Diffuse Anoxic and Traumatic Encephalopathies in Adults , 1988, Journal of clinical neurophysiology : official publication of the American Electroencephalographic Society.

[15]  M. Devor,et al.  Reversible analgesia, atonia, and loss of consciousness on bilateral intracerebral microinjection of pentobarbital , 2001, Pain.

[16]  Jun Lu,et al.  Locus Ceruleus and Anterior Cingulate Cortex Sustain Wakefulness in a Novel Environment , 2010, The Journal of Neuroscience.

[17]  Thomas Benner,et al.  Disconnection of the ascending arousal system in traumatic coma. , 2013, Journal of neuropathology and experimental neurology.

[18]  Ayse Aralasmak,et al.  One-year follow-up in patients with brainstem infarction due to large-artery atherothrombosis , 2015, Neuropsychiatric disease and treatment.

[19]  Y. Katayama,et al.  Activation of pontine cholinergic sites implicated in unconsciousness following cerebral concussion in the cat. , 1984, Science.

[20]  Wade S. Smith,et al.  Initial Glasgow Coma Scale score predicts outcome following thrombolysis for posterior circulation stroke. , 2005, Archives of neurology.

[21]  A. Zeman,et al.  Persistent vegetative state , 1998, The Lancet.

[22]  Nathan S White,et al.  Impaired thalamocortical connectivity in humans during general-anesthetic-induced unconsciousness , 2003, NeuroImage.

[23]  AH Chowdhury,et al.  Assessment of coma and impaired consciousness : A critical review , 2009 .

[24]  T. Sonobe,et al.  H2S induced coma and cardiogenic shock in the rat: Effects of phenothiazinium chromophores , 2015, Clinical toxicology.

[25]  Beth Jelfs,et al.  Vagus Nerve Stimulation Alters Phase Synchrony of the Anterior Cingulate Cortex and Facilitates Decision Making in Rats , 2016, Scientific Reports.

[26]  Emery N. Brown,et al.  Optogenetic activation of dopamine neurons in the ventral tegmental area induces reanimation from general anesthesia , 2016, Proceedings of the National Academy of Sciences.

[27]  M. Steriade,et al.  Reticularis thalami neurons revisited: activity changes during shifts in states of vigilance , 1986, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[28]  Caroline Schnakers,et al.  Non-Invasive Ultrasonic Thalamic Stimulation in Disorders of Consciousness after Severe Brain Injury: A First-in-Man Report , 2016, Brain Stimulation.

[29]  W. Nauta,et al.  Hypothalamic regulation of sleep in rats; an experimental study. , 1946, Journal of neurophysiology.

[30]  Jun Lu,et al.  GABAergic parafacial zone is a medullary slow–wave–sleep promoting center , 2014, Nature Neuroscience.

[31]  Christelle Anaclet,et al.  Basal forebrain control of wakefulness and cortical rhythms , 2015, Nature Communications.

[32]  B. Chenuel,et al.  Immediate and Long-Term Outcome of Acute H2S Intoxication Induced Coma in Unanesthetized Rats: Effects of Methylene Blue , 2015, PloS one.

[33]  S Laureys,et al.  Restoration of thalamocortical connectivity after recovery from persistent vegetative state , 2000, The Lancet.

[34]  M. Kahana,et al.  Comparison of spectral analysis methods for characterizing brain oscillations , 2007, Journal of Neuroscience Methods.

[35]  Jaeseung Jeong,et al.  Frequency distribution of causal connectivity in rat sensorimotor network: resting-state fMRI analyses. , 2013, Journal of neurophysiology.

[36]  Donald H. Lee,et al.  Disruptions of functional connectivity in the default mode network of comatose patients , 2012, Neurology.

[37]  F Joó,et al.  Global Cerebral Ischemia Associated with Cardiac Arrest in the Rat: I. Dynamics of Early Neuronal Changes , 1992, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[38]  Barbara E. Jones,et al.  Effects of locus coeruleus lesions upon cerebral monoamine content, sleep-wakefulness states and the response to amphetamine in the cat , 1977, Brain Research.

[39]  Ramona Hicks,et al.  Therapy development for diffuse axonal injury. , 2013, Journal of neurotrauma.

[40]  Christelle Anaclet,et al.  Activation of the GABAergic Parafacial Zone Maintains Sleep and Counteracts the Wake-Promoting Action of the Psychostimulants Armodafinil and Caffeine , 2018, Neuropsychopharmacology.

[41]  J. Giacino,et al.  The JFK Coma Recovery Scale-Revised: measurement characteristics and diagnostic utility. , 2004, Archives of physical medicine and rehabilitation.

[42]  R. Turner,et al.  Eigenvector Centrality Mapping for Analyzing Connectivity Patterns in fMRI Data of the Human Brain , 2010, PloS one.

[43]  G Lohmann,et al.  LIPSIA--a new software system for the evaluation of functional magnetic resonance images of the human brain. , 2001, Computerized medical imaging and graphics : the official journal of the Computerized Medical Imaging Society.

[44]  Mathias Hoehn,et al.  Current status of functional MRI on small animals: application to physiology, pathophysiology, and cognition , 2007, NMR in biomedicine.

[45]  Michael R. Dashwood,et al.  Endothelin-1 as a neuropeptide: neurotransmitter or neurovascular effects? , 2009, Journal of Cell Communication and Signaling.

[46]  L. Hochberg,et al.  Early detection of consciousness in patients with acute severe traumatic brain injury , 2017, Brain : a journal of neurology.

[47]  L. Schramm,et al.  Behavior of chronically decerebrated kittens. , 1974, Experimental neurology.

[48]  Steven Laureys,et al.  Consciousness in humans and non-human animals: recent advances and future directions , 2013, Front. Psychol..

[49]  Daniel J. Thengone,et al.  Local changes in network structure contribute to late communication recovery after severe brain injury , 2016, Science Translational Medicine.

[50]  Y. Katayama,et al.  Coma induced by cholinergic activation of a restricted region in the pontine reticular formation--a model of reversible forms of coma. , 1986, Neurologia medico-chirurgica.

[51]  Emery N. Brown,et al.  Local cortical dynamics of burst suppression in the anaesthetized brain , 2013, Brain : a journal of neurology.

[52]  R. Neumar,et al.  Outcome Model of Asphyxial Cardiac Arrest in Rats , 1995, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[53]  C. Economo.  SLEEP AS A PROBLEM OF LOCALIZATION , 1930 .

[54]  D. Ingvar,et al.  Electrical activity of isolated cortex in the unanesthetized cat with intact brain stem. , 1955, Acta physiologica Scandinavica.

[55]  D J Bell,et al.  The decerebrate rat , 1933, The Journal of physiology.

[56]  N V Thakor,et al.  Early electrophysiological and histologic changes after global cerebral ischemia in rats , 2000, Movement disorders : official journal of the Movement Disorder Society.

[57]  C. E. Henry,et al.  Suppression-burst activity from isolated cerebral cortex in man. , 1952, Electroencephalography and clinical neurophysiology.

[58]  A. A. Parsons,et al.  A New Primate Model of Focal Stroke: Endothelin-1—Induced Middle Cerebral Artery Occlusion and Reperfusion in the Common Marmoset , 2004, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[59]  J. Hobson,et al.  THE EFFECTS OF CHRONIC BRAIN-STEM LESIONS ON CORTICAL AND MUSCULAR ACTIVITY DURING SLEEP AND WAKING IN THE CAT. , 1965, Electroencephalography and clinical neurophysiology.

[60]  G. Glover,et al.  Respiration‐induced B0 fluctuations and their spatial distribution in the human brain at 7 Tesla , 2002, Magnetic resonance in medicine.

[61]  R W Cox,et al.  AFNI: software for analysis and visualization of functional magnetic resonance neuroimages. , 1996, Computers and biomedical research, an international journal.

[62]  Priyattam J Shiromani,et al.  Effects of Saporin-Induced Lesions of Three Arousal Populations on Daily Levels of Sleep and Wake , 2007, The Journal of Neuroscience.

[63]  M. Raichle,et al.  Rat brains also have a default mode network , 2012, Proceedings of the National Academy of Sciences.

[64]  Steven Laureys,et al.  Practice Guideline Update Recommendations Summary: Disorders of Consciousness: Report of the Guideline Development, Dissemination, and Implementation Subcommittee of the American Academy of Neurology; the American Congress of Rehabilitation Medicine; and the National Institute on Disability, Indepen , 2018, Archives of physical medicine and rehabilitation.

[65]  E. Arrigoni,et al.  Neural Circuitry of Wakefulness and Sleep , 2017, Neuron.

[66]  Alfonso Nieto-Castanon,et al.  Functional networks reemerge during recovery of consciousness after acute severe traumatic brain injury , 2018, Cortex.

[67]  Steven Laureys,et al.  Disorders of consciousness after acquired brain injury: the state of the science , 2014, Nature Reviews Neurology.

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

[69]  Eugen Trinka,et al.  EEG Patterns in Hypoxic Encephalopathies (Post–Cardiac Arrest Syndrome): Fluctuations, Transitions, and Reactions , 2013, Journal of clinical neurophysiology : official publication of the American Electroencephalographic Society.

[70]  H. Michalek,et al.  Transient global brain hypoxia-ischemia in adult rats: Neuronal damage, glial proliferation, and alterations in inositol phospholipid hydrolysis , 1997, Neurochemistry International.

[71]  J. McGaughy,et al.  Cognitive control and the anterior cingulate cortex: How conflicting stimuli affect attentional control in the rat , 2015, Journal of Physiology-Paris.

[72]  Anthony G. Hudetz,et al.  Differential Role of Prefrontal and Parietal Cortices in Controlling Level of Consciousness , 2018, Current Biology.

[73]  J. Brierley,et al.  A New Model of Bilateral Hemispheric Ischemia in the Unanesthetized Rat , 1979, Stroke.

[74]  M. Devor,et al.  Cerebral Activity during the Anesthesia-Like State Induced by Mesopontine Microinjection of Pentobarbital , 2009, The Journal of Neuroscience.

[75]  C. Blanco-Centurion,et al.  Insomnia following hypocretin2-saporin lesions of the substantia nigra , 2006, Neuroscience.

[76]  Sergio Schinelli,et al.  Pharmacology and physiopathology of the brain endothelin system: an overview. , 2006, Current medicinal chemistry.

[77]  B. Jennett,et al.  Assessment of coma and impaired consciousness. A practical scale. , 1974, Lancet.

[78]  F. Hopp,et al.  Improved method of canine decerebration. , 1998, Journal of applied physiology.

[79]  Seiji Nishino,et al.  Basal forebrain circuit for sleep-wake control , 2015, Nature Neuroscience.

[80]  Mircea Steriade.  Brainstem activation of thalamocortical systems , 1999, Brain Research Bulletin.

[81]  O. Hommes,et al.  Recurrent Prolonged Coma Due to Basilar Artery Migraine. A Case Report. , 1991, Headache.

[82]  H. Kurihara,et al.  A novel peptide vasoconstrictor, endothelin, is produced by vascular endothelium and modulates smooth muscle Ca2+ channels , 1988, Journal of hypertension. Supplement : official journal of the International Society of Hypertension.

[83]  A. Damasio,et al.  Consciousness and the brainstem , 2001, Cognition.

[84]  Laura D. Lewis,et al.  Thalamic reticular nucleus induces fast and local modulation of arousal state , 2015, eLife.

[85]  E. Brown,et al.  Thalamocortical model for a propofol-induced α-rhythm associated with loss of consciousness , 2010, Proceedings of the National Academy of Sciences.

[86]  Lippincott Williams Wilkins,et al.  Practice guideline update recommendations summary: Disorders of consciousness , 2019, Neurology.

[87]  Steven Laureys,et al.  Testing Proposed Neuronal Models of Effective Connectivity Within the Cortico‐basal Ganglia‐thalamo‐cortical Loop During Loss of Consciousness , 2016, Cerebral cortex.

[88]  Klaus Scheffler,et al.  Identifying Respiration-Related Aliasing Artifacts in the Rodent Resting-State fMRI , 2018, Front. Neurosci..

[89]  M. Dashwood,et al.  Endothelin‐1: Contractile Responses and Autoradiographic Localization of Receptors in Rabbit Blood Vessels , 1989, Journal of cardiovascular pharmacology.

[90]  W. B. Knowles,et al.  Behavioral and EEG changes following chronic brain stem lesions in the cat. , 1950, Electroencephalography and clinical neurophysiology.

[91]  Wei-Min Qu,et al.  The rostromedial tegmental nucleus is essential for non-rapid eye movement sleep , 2018, PLoS biology.

[92]  P. Safar,et al.  Journal of Cerebral Blood Flow and Metabolism Mild Cerebral Hypothermia during and after Cardiac Arrest Improves Neurologic Outcome in Dogs , 2022 .

[93]  Barbara E. Jones,et al.  Neurotoxic lesions of the dorsolateral pontomesencephalic tegmentum-cholinergic cell area in the cat. II. Effects upon sleep-waking states , 1988, Brain Research.

[94]  Claude Matuchansky.  Diagnosis of reversible causes of coma , 2015, The Lancet.

[95]  Li-Na Zhang,et al.  Orexin-A facilitates emergence of the rat from isoflurane anesthesia via mediation of the basal forebrain , 2016, Neuropeptides.

[96]  Anita Lüthi.  Sleep: Switching Off the Off-Switch , 2016, Current Biology.

[97]  Seth R. Jones,et al.  Resting‐state functional connectivity of the rat brain , 2008, Magnetic resonance in medicine.

[98]  R. B. Ebitz,et al.  Neuronal Activity in Primate Dorsal Anterior Cingulate Cortex Signals Task Conflict and Predicts Adjustments in Pupil-Linked Arousal , 2015, Neuron.

[99]  Andrzej Loesch.  Localisation of endothelin-1 and its receptors in vascular tissue as seen at the electron microscopic level. , 2005, Current vascular pharmacology.

[100]  D. Heistad,et al.  Role of the basilar artery in regulation of blood flow to the brain stem in rats. , 1991, Stroke.

[101]  Alexis Roche,et al.  Natural history of recovery from brain injury after prolonged disorders of consciousness: outcome of patients admitted to inpatient rehabilitation with 1-4 year follow-up. , 2009, Progress in brain research.

[102]  N. Schiff,et al.  Cognitive enhancement with central thalamic electrical stimulation , 2006, Proceedings of the National Academy of Sciences.

[103]  Miklós Emri,et al.  Voxel-Wise Motion Artifacts in Population-Level Whole-Brain Connectivity Analysis of Resting-State fMRI , 2014, PloS one.

[104]  B. Krämer,et al.  Role of endothelin in hypertension , 2004, Journal of Molecular Medicine.

[105]  C. Hawkes,et al.  The electroencephalogram in the “locked‐in” syndrome , 1974, Neurology.

[106]  Jun Lu,et al.  Reassessment of the structural basis of the ascending arousal system , 2011, The Journal of comparative neurology.

[107]  Ruben G. L. Real,et al.  Traumatic brain injury: integrated approaches to improve prevention, clinical care, and research , 2017, The Lancet Neurology.

[108]  D. Simons,et al.  Thalamocortical Dysfunction and Thalamic Injury after Asphyxial Cardiac Arrest in Developing Rats , 2012, The Journal of Neuroscience.

[109]  Á. Pascual-Leone,et al.  A human brain network derived from coma-causing brainstem lesions , 2016, Neurology.

[110]  D B Lindsley,et al.  Effect upon the EEG of acute injury to the brain stem activating system. , 1949, Electroencephalography and clinical neurophysiology.

[111]  Joseph P Broderick,et al.  Basilar Occlusion Syndromes , 2015, The Neurohospitalist.

[112]  G. Galardi,et al.  Prognostic value of standard EEG in traumatic and non-traumatic disorders of consciousness following coma , 2010, Clinical Neurophysiology.

[113]  J. Gold,et al.  Relationships between Pupil Diameter and Neuronal Activity in the Locus Coeruleus, Colliculi, and Cingulate Cortex , 2016, Neuron.

[114]  Zuoren Wang,et al.  Persistent Neuronal Activity in Anterior Cingulate Cortex Correlates with Sustained Attention in Rats Regardless of Sensory Modality , 2017, Scientific Reports.

[115]  A. Borbély,et al.  The effect of sleep deprivation on sleep in rats with suprachiasmatic lesions , 1983, Neuroscience Letters.

[116]  C. Chapman,et al.  Partial antagonism of propofol anaesthesia by physostigmine in rats is associated with potentiation of fast (80-200 Hz) oscillations in the thalamus. , 2013, British journal of anaesthesia.

[117]  S. Leibowitz,et al.  Hypothalamic control of energy balance: different peptides, different functions , 2004, Peptides.

[118]  Nitish V. Thakor,et al.  Quantitative EEG and neurological recovery with therapeutic hypothermia after asphyxial cardiac arrest in rats , 2006, Brain Research.

[119]  Terrence J. Sejnowski,et al.  Brain-state dependent astrocytic Ca2+ signals are coupled to both positive and negative BOLD-fMRI signals , 2018, Proceedings of the National Academy of Sciences.

[120]  G. Boylan,et al.  Global suppression of electrocortical activity in unilateral perinatal thalamic stroke , 2014, Developmental medicine and child neurology.

[121]  Richard Rogers,et al.  Cortical and Subcortical Connectivity Changes during Decreasing Levels of Consciousness in Humans: A Functional Magnetic Resonance Imaging Study using Propofol , 2010, The Journal of Neuroscience.

[122]  G. Moruzzi,et al.  Brain stem reticular formation and activation of the EEG. , 1949, Electroencephalography and clinical neurophysiology.

[123]  Mami Okada,et al.  Development of an experimental rat model of hyperammonemic encephalopathy and evaluation of the effects of rifaximin. , 2016, European journal of pharmacology.

[124]  F. Plum,et al.  Behavioural improvements with thalamic stimulation after severe traumatic brain injury , 2008, Nature.

[125]  C. Saper,et al.  Organization of cerebral cortical afferent systems in the rat. II. Hypothalamocortical projections , 1985, The Journal of comparative neurology.

[126]  Yan Xu,et al.  A reproducible model of circulatory arrest and remote resuscitation in rats for NMR investigation. , 1998, Stroke.

[127]  Thomas E. Scammell,et al.  The sleep switch: hypothalamic control of sleep and wakefulness , 2001, Trends in Neurosciences.

[128]  N. Thakor,et al.  Vulnerability of the thalamic somatosensory pathway after prolonged global hypoxic–ischemic injury , 2002, Neuroscience.

[129]  N. Schiff.  Recovery of consciousness after brain injury: a mesocircuit hypothesis , 2010, Trends in Neurosciences.

[130]  Kumiko Okamura,et al.  Glasgow Coma Scale flow chart: a beginner's guide. , 2014, British journal of nursing.

[131]  Synek Vm,et al.  Prognostically important EEG coma patterns in diffuse anoxic and traumatic encephalopathies in adults. , 1988 .

[132]  C. Saper,et al.  Hypothalamic regulation of sleep and circadian rhythms , 2005, Nature.

[133]  J. Bogousslavsky,et al.  Heralding manifestations of basilar artery occlusion with lethal or severe stroke , 2003, Journal of neurology, neurosurgery, and psychiatry.

[134]  Hassan Azari,et al.  Endothelin-1 induced middle cerebral artery occlusion model for ischemic stroke with laser Doppler flowmetry guidance in rat. , 2013, Journal of visualized experiments : JoVE.

[135]  Florin Amzica,et al.  Human Brain Activity Patterns beyond the Isoelectric Line of Extreme Deep Coma , 2013, PloS one.

[136]  Bruce R. Rosen,et al.  Ultra-Slow Single-Vessel BOLD and CBV-Based fMRI Spatiotemporal Dynamics and Their Correlation with Neuronal Intracellular Calcium Signals , 2018, Neuron.