Name: Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice (Video) | JoVE Journal
Uploaded: 2020-06-11T15:00:00.000+00:00
Duration: 9 min 29 s
Description: Learn continuous video EEG recording in neonatal mice during hypoxia-ischemia to study seizure mechanisms. This method enables correlation of electrographic activity with behavioral events for therapeutic development.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

We acknowledge the following funding sources: NIH NINDS &#8211; K08NS101122 (JB), R01NS040337 (JK), R01NS044370 (JK), University of Virginia School of Medicine (JB).

All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Virginia.
1. Electrode building/cable building
<ol>
	<li>Use a unipolar insulated stainless-steel wire (0.005&#8221; bare diameter, 0.008&#8221; coated) to make an electrode that is connected with a female socket connector (female receptacle connector 0.079).</li>
	<li>Use a special custom-made cable to connect animals to the amplifier.
	<ol>
		<li>Attach a male 4-pin connector (Male connector 0.079&#8221;) to 4 channel unity gain impedance matching operational amplifier (op-amp). Attach a 10K resistor to the wires that connect to the 9 V battery. A ground wire not connected to the op-amp acts as the midpoint of the battery.</li>
		<li>Connect one end of the cable (AWG, 0.012&#8221; OD) to the op-amp and connect the other end of the cable to the amplifier.</li>
	</ol>
	</li>
</ol>
2. Electrode implantation surgery
<ol>
	<li>Anesthetize the pup (postnatal day 9) with 4-5% isoflurane in a downward flow hood. Prior to the start of the procedure, inject the pups with bupivacaine (0.02-0.05 mL, 0.25% subcutaneous local infiltration).</li>
	<li>Once the animal is immobile, transfer to a stereotactic stage with a nose cone. Use the reverse side of the ear bar as it is soft to hold the head steady. At this age, pups do not have a fully developed ear to use the pointed end of the ear bar.</li>
	<li>Turn down the flow of isoflurane and maintain it at 2.5-3%. Keep an eye on steady breathing of the pup throughout the surgery procedure. Pinch the tail to check pain response and then proceed to incision.</li>
	<li>Sterilize the incision area on the skull with betadine and alcohol (3 cycles of alternating iodine and 70% ethanol). Drape the surrounding body part such that the incision region is visible.</li>
	<li>Open the scalp anterior-posterior from slightly above the eyes and retract approximately 0.5&#160;cm of skin. Reposition the mouse head on the stereotaxic stage so that the skin pulls outward exposing skull.</li>
	<li>Apply hydrogen peroxide on the skull using a cotton swab and scrape the skull clean using a scalpel blade. The skull is very soft; exercise caution while scraping.</li>
	<li>Apply one drop (approximately 50 &#181;L) of adhesive and spread it around the exposed skull area using its applicator. Expose to UV light for 40 s to set the adhesive.</li>
	<li>Measure the coordinates using the exposed bregma as the reference. Implant electrodes bilaterally in the CA1 region of hippocampus [-3.5 mm Dorsal-Ventral (DV), &#177;2 mm Medial-Lateral (ML), -1.75 mm Deep (D)] and bilaterally in the parietal cortex [-1.22 mm DV, &#177;0.5 mm ML, -1 mm D] and a reference electrode in the cerebellum17. Use a 32 G needle to create a hole at the marked region.</li>
	<li>Clean the blood from the surface of the skull. Lower electrodes attached to the female socket connector into the brain with the help of the stereotaxic arm and fix in place with dental acrylic. Implant the electrode in the brain. The socket connector headset sits on top of the skull glued together by dental acrylic.</li>
	<li>Inject ketoprofen (5 mg/kg) subcutaneously in the interscapular region once the electrode is fixed. Place the pups back with the mother. 
	NOTE: Introduce half of the litter with the headset at once to the mother rather than introducing them one at a time. This will prevent from mother damaging the pup&#8217;s headset.</li>
</ol>
3. EEG setup and recording (baseline/pre-injury)
<ol>
	<li>After 24 h of recovery after electrode implantation, place each animal in a heated (37 &#176;C) custom-made Plexiglas chamber for EEG recording. This chamber will also serve as a hypoxia chamber.</li>
	<li>Connect pups in the chamber to a video-EEG monitoring system via a flexible cable (custom made op-amp cable). 
	NOTE: With the headset in place, the mice are freely mobile and do not exhibit any differences in behavior. Once attached to the electrode wires, the wires must be adjusted within the chamber tether in order to provide the right amount of slack so that the pup can freely move throughout the chamber.</li>
	<li>Digitize the EEG data at 1000 Hz with 1K gain using a grass amplifier. Review the EEG signal (band pass filter between 3-70 Hz) later using software (e.g., LabChart Pro).</li>
	<li>Record a pre-injury baseline EEG for 30 minutes prior to disconnecting animals for carotid artery ligation procedure.</li>
</ol>
4. Left carotid artery ligation
<ol>
	<li>Anesthetize the pup (postnatal day 10) with 4-5% isoflurane in a downward flow hood and place them on specially arranged setup on a waterbath pad. Position the animal supine and secure the forelimbs with paper tape.
	<ol>
		<li>Lower the flow of isoflurane to 2-3%. Pinch the tail for pain response and monitor breathing throughout procedure.</li>
	</ol>
	</li>
	<li>Sterilize the incision area (between mandible and the clavicle) on the left side of the neck with betadine and alcohol (3 cycles of alternating iodine and 70% ethanol).</li>
	<li>Make an approximately 1 cm long incision on the left side of the neck using microscissors. Using a dissecting microscope, carefully retract the subcutaneous tissue and skin to expose the carotid artery. Take care to identify the vagus nerve (running lateral to the artery) and delicately separate and retract it from the artery.</li>
	<li>Thread a 5 cm long sterile silk suture under the artery using microforceps. Tie a double knotted suture around the artery to occlude flow.</li>
	<li>Cut the excess suture and close the exposed artery by pulling back the subcutaneous tissue and skin. Use vet bond to seal the incision.</li>
	<li>Place the animal back on continuous EEG monitoring in a chamber at room temperature, which is placed on a warming mattress. Take spot infrared temperature checks of pup core temperature to avoid opening the chamber. Allow the animal to recover for 1 h prior to hypoxia.</li>
</ol>
5. EEG and hypoxia
<ol>
	<li>Continuously monitor FiO2 (fraction of inspired oxygen) within the chamber via an oxygen monitor.</li>
	<li>Flush the chamber with 60 L/min of 100% N2 and 0.415 L/min for 100% O2. Once the oxygen saturation in the chamber reaches 12%, decrease the N2 flow to 10 L/min while keeping the O2 flow unchanged. With small adjustments, maintain the FiO2 at 8% for 45 min.</li>
	<li>After 45 min of hypoxia exposure, return FiO2 to 21%.</li>
	<li>Have pups recover in chamber and monitor on EEG for 2 h post-hypoxia.</li>
	<li>After completion of recording period, disconnect mice from EEG recording and return to the mother.</li>
</ol>
6. EEG analysis
<ol>
	<li>Analyze the EEG file with video in LabChart Pro. Have a blinded researcher mark the EEG for seizures and background patterns17. Seizures are defined as an electrographic event lasting greater than 10 seconds with high frequency rhythmic sharp wave discharges (&#8805;3x baseline) with clear evolution17.</li>
	<li>Have a second blinded researcher review marked events at random for agreement.</li>
	<li>Review associated video for each marked electrographic event and analyze according to the neonatal rodent behavioral seizure score16. Briefly, this score ranges from 0-6 (immobility to severe tonic-clonic behavior). To further characterize seizure semiology, analyze behavior for laterality (multifocal/bilateral movements vs. focal/unilateral vs. mixed).</li>
	<li>Create a power spectrogram. Use a Fast Fourier Transform with a Cosine-Bell data window with a size of 1024 data points. Create a smooth x-axis in the spectrogram with the help of a window overlap of 87.5%. Express the power as &#181;V218.</li>
</ol>

<ol><li>Vasudevan, C., Levene, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epidemiology+and+aetiology+of+neonatal+seizures%5BTitle%5D)+AND+%22Seminars+in+Fetal+%26+Neonatal+Medicine%22%5BJournal%5D)">Epidemiology and aetiology of neonatal seizures.</a> Seminars in Fetal & Neonatal Medicine. , (2013).</li>
<li>Volpe, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neonatal+Seizures%5BTitle%5D)+AND+%22Volpe%27s+Neurology+of+the+Newborn%22%5BJournal%5D)">Neonatal Seizures.</a> Volpe's Neurology of the Newborn. , Elsevier. 275-321 (2018).</li>
<li>Shankaran, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Network+EKSNNR.+Childhood+outcomes+after+hypothermia+for+neonatal+encephalopathy%5BTitle%5D)+AND+%22New+England+Journal+of+Medicine%22%5BJournal%5D)">Network EKSNNR. Childhood outcomes after hypothermia for neonatal encephalopathy.</a> New England Journal of Medicine. 366 (22), 2085-2092 (2012).</li>
<li>Pappas, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cognitive+outcomes+after+neonatal+encephalopathy%5BTitle%5D)+AND+%22Pediatrics%22%5BJournal%5D)">Cognitive outcomes after neonatal encephalopathy.</a> Pediatrics. 135 (3), 624-634 (2015).</li>
<li>van Schie, P. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Long-term+motor+and+behavioral+outcome+after+perinatal+hypoxic-ischemic+encephalopathy%5BTitle%5D)+AND+%22European+Journal+of+Paediatric+Neurology%22%5BJournal%5D)">Long-term motor and behavioral outcome after perinatal hypoxic-ischemic encephalopathy.</a> European Journal of Paediatric Neurology. 19 (3), 354-359 (2015).</li>
<li>Rensing, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Longitudinal+analysis+of+developmental+changes+in+electroencephalography+patterns+and+sleep-wake+states+of+the+neonatal+mouse%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Longitudinal analysis of developmental changes in electroencephalography patterns and sleep-wake states of the neonatal mouse.</a> PLoS One. 13 (11), 1-17 (2018).</li>
<li>Rice, J. E., Vannucci, R. C., Brierley, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+influence+of+immaturity+on+hypoxic-ischemic+brain+damage+in+the+rat%5BTitle%5D)+AND+%22Annals+of+Neurology%22%5BJournal%5D)">The influence of immaturity on hypoxic-ischemic brain damage in the rat.</a> Annals of Neurology. 9 (2), 131-141 (1981).</li>
<li>Burnsed, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hypoxia-ischemia+and+therapeutic+hypothermia+in+the+neonatal+mouse+brain--a+longitudinal+study%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Hypoxia-ischemia and therapeutic hypothermia in the neonatal mouse brain--a longitudinal study.</a> PLoS One. 10 (3), 0118889(2015).</li>
<li>Semple, B. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Brain+development+in+rodents+and+humans%3A+Identifying+benchmarks+of+maturation+and+vulnerability+to+injury+across+species%5BTitle%5D)+AND+%22Progress+in+Neurobiology%22%5BJournal%5D)">Brain development in rodents and humans: Identifying benchmarks of maturation and vulnerability to injury across species.</a> Progress in Neurobiology. , 1-16 (2013).</li>
<li>Comi, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Gabapentin+neuroprotection+and+seizure+suppression+in+immature+mouse+brain+ischemia%5BTitle%5D)+AND+%22Pediatric+Research%22%5BJournal%5D)">Gabapentin neuroprotection and seizure suppression in immature mouse brain ischemia.</a> Pediatric Research. 64 (1), 81-85 (2008).</li>
<li>Comi, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+new+model+of+stroke+and+ischemic+seizures+in+the+immature+mouse%5BTitle%5D)+AND+%22Pediatric+Neurology%22%5BJournal%5D)">A new model of stroke and ischemic seizures in the immature mouse.</a> Pediatric Neurology. 31 (4), 254-257 (2004).</li>
<li>Kadam, S. D., White, A. M., Staley, K. J., Dudek, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Continuous+Electroencephalographic+Monitoring+with+Radio-Telemetry+in+a+Rat+Model+of+Perinatal+Hypoxia-Ischemia+Reveals+Progressive+Post-Stroke+Epilepsy%5BTitle%5D)+AND+%22Journal+of+Neuroscience%22%5BJournal%5D)">Continuous Electroencephalographic Monitoring with Radio-Telemetry in a Rat Model of Perinatal Hypoxia-Ischemia Reveals Progressive Post-Stroke Epilepsy.</a> Journal of Neuroscience. 30 (1), 404-415 (2010).</li>
<li>Burnsed, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neuronal+Circuit+Activity+during+Neonatal+Hypoxic+-+Ischemic+Seizures+in+Mice%5BTitle%5D)+AND+%22Annals+of+Neurology%22%5BJournal%5D)">Neuronal Circuit Activity during Neonatal Hypoxic - Ischemic Seizures in Mice.</a> Annals of Neurology. 86, 927-938 (2019).</li>
<li>Sampath, D., White, A. M., Raol, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characterization+of+neonatal+seizures+in+an+animal+model+of+hypoxic-ischemic+encephalopathy%5BTitle%5D)+AND+%22Epilepsia%22%5BJournal%5D)">Characterization of neonatal seizures in an animal model of hypoxic-ischemic encephalopathy.</a> Epilepsia. 55 (7), 985-993 (2014).</li>
<li>Sampath, D., Valdez, R., White, A. M., Raol, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Anticonvulsant+effect+of+flupirtine+in+an+animal+model+of+neonatal+hypoxic-ischemic+encephalopathy%5BTitle%5D)+AND+%22Neuropharmacology%22%5BJournal%5D)">Anticonvulsant effect of flupirtine in an animal model of neonatal hypoxic-ischemic encephalopathy.</a> Neuropharmacology. 123, 126-135 (2017).</li>
<li>Kang, S. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((and+sex-dependent+susceptibility+to+phenobarbital-resistant+neonatal+seizures%3A+role+of+chloride+co-transporters%5BTitle%5D)+AND+%22Frontiers+in+Cellular+Neuroscience%22%5BJournal%5D)">and sex-dependent susceptibility to phenobarbital-resistant neonatal seizures: role of chloride co-transporters.</a> Frontiers in Cellular Neuroscience. 9, 1-16 (2015).</li>
<li>Zanelli, S., Goodkin, H. P., Kowalski, S., Kapur, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Impact+of+transient+acute+hypoxia+on+the+developing+mouse+EEG%5BTitle%5D)+AND+%22Neurobiology+of+Disease%22%5BJournal%5D)">Impact of transient acute hypoxia on the developing mouse EEG.</a> Neurobiology of Disease. 68, 37-46 (2014).</li>
<li>Lewczuk, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((EEG+and+behavior+patterns+during+experimental+status+epilepticus%5BTitle%5D)+AND+%22Epilepsia%22%5BJournal%5D)">EEG and behavior patterns during experimental status epilepticus.</a> Epilepsia. 59 (2), 369-380 (2017).</li>
<li>Wu, D., Martin, L. J., Northington, F. J., Zhang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Oscillating+gradient+diffusion+MRI+reveals+unique+microstructural+information+in+normal+and+hypoxia-ischemia+injured+mouse+brains%5BTitle%5D)+AND+%22Magnetic+Resonance+in+Medicine%22%5BJournal%5D)">Oscillating gradient diffusion MRI reveals unique microstructural information in normal and hypoxia-ischemia injured mouse brains.</a> Magnetic Resonance in Medicine. 72 (5), 1366-1374 (2014).</li>
</ol>

We have presented a model for continuous video-EEG monitoring in neonatal mice during hypoxic-ischemic seizures. Video analysis in conjunction with EEG allows characterization of seizure semiology. Analysis of EEG allows for extraction of power spectrograms and background amplitude analysis.
Correct and careful placement of electrodes is crucial in this protocol, as injury during electrode placement or inaccurate placement can significantly affect results. Assessment of normal baseline EEG act...

Hypoxic ischemic encephalopathy (HIE) is a condition that affects 1.5 in 1000 newborns annually and is the most common cause of neonatal seizures1,2. Infants who survive are at risk for various neurological disabilities such as cerebral palsy, intellectual disability, and epilepsy3,4,5.
Animal models play a critical role in understanding and investigating the pathophysiology of hypoxia ischemia and neonatal seizures6,7. A modified Vannucci model is used to induce hypoxia ischemia (HI) on postnatal day 10 (p10)7,8. Mouse pups of this age translate neurologically roughly to the full term human neonate9.
Continuous video electroencephalography (EEG) monitoring used in conjunction with this injury model allows for further understanding and characterization of neonatal hypoxic ischemic seizures. Previous studies have used various methods for analyzing neonatal seizures in rodents, including video recordings, limited EEG recordings and telemetry EEG recordings10,11,12,13,14,15,16. In the following manuscript, we discuss in depth the process of recording continuous video EEG in mouse pups during hypoxia-ischemia. This technique for continuous video EEG monitoring in neonatal mouse pups could be applied to a variety of disease and seizure models.

<table><tbody><tr><td>&lt;strong&gt;SURGERY&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Ball Point Applicator</td><td>Metrex Research</td><td>8300-F</td><td>i-bond applicator</td></tr><tr><td>Cranioplast (Powder/Resin)</td><td>Coltene</td><td>H00383</td><td>Perm Reline/Power</td></tr><tr><td>I-Bond</td><td>Kulzer GmbH, Germany</td><td></td><td></td></tr><tr><td>LOOK Silk Suture</td><td>Surgical Specialities Corporation</td><td>SP115</td><td>LOOK SP115 Black Braided Silk Non absorbable surgical suture</td></tr><tr><td>RS-5168 Botvin Forceps</td><td>Roboz Surgical Instrument</td><td>RS5168</td><td>Forcep for surgery/ligation</td></tr><tr><td>RS-5138 Graefe Forceps</td><td>Roboz Surgical Instrument</td><td>RS5138</td><td>Forcep for surgery/ligation</td></tr><tr><td>UV light for I-Bond</td><td>Blast Lite By First Media</td><td>BL778</td><td>UV ligth for I-bond</td></tr><tr><td>Vannas Microdissecting Scissor</td><td>Roboz Surgical Instrument</td><td>RS5618</td><td>Scissor for ligation</td></tr><tr><td>Vet Bond</td><td>3M Vetbond</td><td>1469SB</td><td>Vet Glue</td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;HYPOXIA&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Hypoxidial</td><td>Starr Life Science</td><td></td><td></td></tr><tr><td>Oxygen sensor</td><td>Medical Products</td><td></td><td>MiniOxI- oxygen analyzer/sensor for hypoxia rig</td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>&lt;strong&gt;EEG RECORDING&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Female receptacle connector 0.079&quot;</td><td>Mill-Max Manufacturing Corp</td><td>832-10-024-10-001000</td><td>Ordered from Digikey</td></tr><tr><td>Grass Amplifier</td><td>Natus Neurology Incorporated</td><td></td><td>Grass Product</td></tr><tr><td>LabChart Pro</td><td>ADI Instruments</td><td></td><td>Software to run the system</td></tr><tr><td>Male Socket Connector 0.079&quot;</td><td>Mill-Max Manufacturing Corp</td><td>833-43-024-20-001000</td><td>Ordered from Digikey</td></tr><tr><td>Operational Amplifier</td><td>Texas Instruments, Dallas, TX, USA</td><td>TLC2274CD</td><td>TLC2274 Quad Low-Noise Rail-to Rail Operational Amplifier</td></tr><tr><td>Operational Amplifier</td><td>Texas Instruments, Dallas, TX, USA</td><td>TLC2272ACDR</td><td>TLC2274 Quad Low-Noise Rail-to Rail Operational Amplifier</td></tr><tr><td>Stainless Steel wire</td><td>A-M Systems</td><td>791400</td><td>0.005&quot; Bare/0.008&quot; Coated 100 ft</td></tr><tr><td>Ultra-Flexible Wire</td><td>McMaster-Carr</td><td>9564T1</td><td>36 Gauze wire of various color</td></tr></tbody></table>

continuous video electroencephalogram during hypoxia-ischemia in neonatal mice

Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

Hypoxia ischemia is the most common cause of neonatal seizures. Animal models are crucial for understanding the mechanisms and physiology underlying neonatal seizures and hypoxia ischemia. This manuscript describes a method for continuous video electroencephalogram (EEG) monitoring in neonatal mice to detect seizures and analyze EEG background during hypoxia ischemia. Use of video and EEG in conjunction allows description of seizure semiology and confirmation of seizures. This method also allows analysis of power spectrograms and EEG background pattern trends over the experimental time period. In this hypoxia ischemia model, the method allows EEG recording prior to injury to obtain a normative baseline and during injury and recovery. Total monitoring time is limited by the inability to separate pups from the mother for longer than four hours. Although, we have used a model of hypoxic-ischemic seizures in this manuscript, this method for neonatal video EEG monitoring could be applied to diverse disease and seizure models in rodents.

Seizure semiology
Neonatal hypoxia-ischemia exposure results in both generalized and focal seizures in mice (Figure 1A-C). Video EEG recordings allow electrographic findings to be correlated to behavior on video. These behaviors were scored using a previously published neonatal rodent behavioral seizure score (BSS)16. In addition to BSS, we categorized events based on whether the behavior was focal/unilateral, bila...

This manuscript describes a method for continuous video EEG recordings using multiple depth electrodes in neonatal mice undergoing hypoxia-ischemia.

Hypoxia ischemia is the most common cause of neonatal seizures. Animal models are crucial for understanding the mechanisms and physiology underlying ...

continuous-video-electroencephalogram-during-hypoxia-ischemia

Research

JoVE Journal

Neuroscience

The postnatal rat model for hypoxic-ischemic brain injury is a well-established model of human neonatal hypoxic ischemic encephalopathy (HIE). In this article, we describe the model of HIE in post-natal rat pups.

The Hypoxic Ischemic Encephalopathy Model of Perinatal Ischemia

Large animal models have good translational values in the examination of physiology and pharmacology of neonatal asphyxia. Using newborn piglets, we develop an experimental protocol to simulate neonatal asphyxia which has advantages of studying the systemic and regional hemodynamics, oxygen transport with biochemical and pathologic pathways and correlations.

A Swine Model of Neonatal Asphyxia

The video demonstrates the procedure for creating a mouse model to study neonatal hypoxic-ischemic brain injury. It outlines the steps for surgically inducing ischemia and subjecting the animal to hypoxia to exacerbate brain damage. Finally, it shows how to assess the resulting brain lesions, ensuring the mouse model is prepared for further scientific investigation.

Establishing a Mouse Model of Neonatal Hypoxic-Ischemic Brain Injury

Source: Ouyang, Y., et al. Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia. J. Vis. Exp. (2015)This video demonstrates a technique to assess cerebral hypoxia injury via simultaneous positron-emission tomography (PET) and magnetic resonance imaging (MRI). An anesthetized mouse undergoes common carotid artery (CCA) ligation, reducing blood flow to one brain hemisphere and predisposing it to hypoxia-induced injury. PET and MRI imaging evaluate glucose metabolism and structural...

Imaging Brain Damage Induced by Cerebral Hypoxia-Ischemia in a Mouse Model

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Here we demonstrate the use of a wireless enabling technology for electroencephalogram (EEG) in neonatal rodent models of human disease. With telemetry, there are no encumbering connections, thus allowing natural behaviors.

Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System

We performed unilateral carotid artery occlusion on postnatal day 7-10 CD-1 mouse pups to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of HI brain injury. We studied neurobehavioral functions in these mice compared to non-operated normal mice.

Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury

Here, we present a protocol to record brain and heart bio signals in mice using simultaneous video, electroencephalography (EEG), and electrocardiography (ECG). We also describe methods to analyze the resulting EEG-ECG recordings for seizures, EEG spectral power, cardiac function, and heart rate variability.

Simultaneous Video-EEG-ECG Monitoring to Identify Neurocardiac Dysfunction in Mouse Models of Epilepsy

Here, we present a noninvasive electrocardiography (ECG) protocol, optimized for early postnatal mice, that does not require the use of anesthetics.

Noninvasive Electrocardiography in the Perinatal Mouse

The present protocol describes the echocardiographic assessment of left ventricular morphology, function, and coronary blood flow in 7-day old neonate mice.

Echocardiographic Characterization of Left Ventricular Structure, Function, and Coronary Flow in Neonate Mice

Learn continuous video EEG recording in neonatal mice during hypoxia-ischemia to study seizure mechanisms. This method enables correlation of electrographic activity with behavioral events for therapeutic development.

Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

Explore More Videos