The authors sincerely thank the Canadian Institutes of Health Research MOP 130258 and the Star Foundation for Children&#8217;s Health Research, along with the Molly Towell Foundation, for the provision of the research facility and financial support. The authors also sincerely thank Dr. Kinkead Richard for manuscript proofreading and advice.

This method required the use of animal subjects, allowed by Laval University Animal Ethics Committee (protocol # 2012-170).
1. Setup and Preparation

<ol>
	<li>Solutions
	<ol>
		<li>Prepare aCSF stock solutions according to the following recipes 7,17. Other recipes with concentration variations are available in the literature. Store stock solutions at 4 &#176;C for up to one month.
		<ol>
			<li>Salt solution: add 75.39 g of NaCl (129 mM final); 2.5 g of KCl (3.35 mM final); 0.81 g of NaH2PO4 (0.58 mM final); 2.33 g of MgCl2 (1.15 mM final); 1.85 g of CaCl2 (1.26 mM). Dissolve in 800 ml of distilled water then fill to 1 L with distillated water.</li>
			<li>Bicarbonate solution: add 17.65 g of NaHCO3 (21 mM final). Dissolve in 900 ml of distillated water then fill to 1 L with distilled water. Variations of the bicarbonate concentration will result in pH variations.</li>
			<li>Glucose solution: add 54.06 g of glucose (30 mM final). Dissolve in 400 ml of distillated water then fill to 500 ml with distilled water.</li>
		</ol>
		</li>
		<li>Prepare aCSF by diluting 100 ml of salt solution, 100 ml of bicarbonate solution, and 50 ml of glucose solution in 1 L of distilled water. Store at 4 &#176;C until use.</li>
		<li>Prepare a glass electrode by warming and stretching a glass tube until it breaks. Sand down the tip before use. The electrode can be reused many time as long as it stays clean.</li>
	</ol>
	</li>
	<li>Experimental Setup 
	Note: The set-up details are presented in Figure 2.
	<ol>
		<li>Turn on the amplifier, moving averager, data acquisition system, temperature controller and pump. Check the signal amplification (gain = 10,000), the filtration (low threshold, 10 Hz; high threshold, 5 kHz), and the sampling rate of the analog to digital conversion of the raw signal (2.5 kHz).</li>
		<li>Fill a bottle with aCSF and bubble it with carbogen (95% O2, 5% CO2). Induce a bubbled- and warmed-aCSF flow in the recording chamber (volume 5 ml) at 4 ml/min. Hold the temperature in the recording chamber at 26 (&#177; 1) &#176;C. pH should be 7.4 in these standard conditions.</li>
		<li>Keep 50 ml of RT and carbogen-bubbled aCSF in a 50 ml syringe near the dissection chamber.</li>
		<li>Turn on the computer and start the recording software.</li>
	</ol>
	</li>
</ol>
2. Dissection
<ol>
	<li>Weigh and visually determine the sex of the animal (males have a longer&#160;ano-genital distance, while females have a short&#160;ano-genital distance; males genitals are often dark while female genitals are pink).</li>
	<li>Anesthetize the newborn rodents by one of the following options: cryoanesthesia (completely immerse the animal in ice for 4 &#8211; 5 min 18), injection (equitensine at 4 ml/kg) 19 or inhalation of volatile anesthetics (isoflurane 20 or ether 21). Confirm an adequate plane of anesthesia by the absence of a paw withdrawal reflex.</li>
	<li>Place the animal on the bench, ventral face down. Section the rostral part of the head coronally with a scalpel at the bregma level (visible through the skin and skull). Perform this step immediately after the animal is anesthetized.</li>
	<li>Section the body coronally with a scalpel under the anterior members .</li>
	<li>Remove skin, muscles, fatty tissues and viscus with surgical and thin scissors and pliers. Since bones are soft at this age, be cautious to not damage the nervous system. Perform this step on the bench.</li>
	<li>Place the preparation in the dissection chamber. Use the aCSF stored in the syringe to oxygenate the preparation. From this point to the end of the recording, use a microscope.</li>
	<li>On the dorsal face of the preparation, cut the skull and vertebras from the rostral to the caudal part along the medium axis with surgical and thin scissors and pliers. Open the cut skull and vertebras in order to expose the nervous tissue. Use the aCSF strored in the syringe to oxygenate the preparation.</li>
	<li>With the pliers, remove the Arachnoid membrane, a thin tissue covering the surface of the nervous tissue. Retain the pia mater and blood vessels against the nervous tissue. Use the aCSF stored in the syringe to oxygenate the preparation.</li>
	<li>Place the preparation with the dorsal face down, and carefully bring down the brainstem and spinal cord by cutting the nerves and connective tissue with the scissors while holding the preparation in place. A dorsal approach is also possible. Keep the roots and nerves as long as possible. Remove the bones to isolate the brainstem and spinal cord. Use the aCSF stored in the syringe to oxygenate the preparation.</li>
	<li>Remove the cerebellum and remaining rostral structures by sectioning them with the scalpel. Use the aCSF stored in the needle to oxygenate the preparation.</li>
	<li>Optionally depending on experimental design, remove the pons with the scalpel by a coronal section anterior to the inferior cerebellar artery 22. Note that keeping the whole pons will slow down the rhythm in rats and fully inhibit it in mice. Preparations that include only the caudal part of the pons display a respiratory-like activity with a stable frequency at the C4 root.</li>
</ol>
3. Recording
<ol>
	<li>Place the preparation in the recording chamber, ventral face up. Fix the preparation with pins in the lowest part of the spinal cord and rostral-most part of the brainstem.</li>
	<li>Using a syringe linked to the electrode by its needle, induce a depression in the electrode (glass tip diameter: 150 - 225 &#181;m) by pulling out the syringe piston, in order to partially fill the electrode with aCSF.</li>
	<li>Using a micromanipulator, carefully place the electrode close to one of the fourth ventral roots. Other ventral roots could also present a respiratory-like activity (e.g., XII cranial nerve, C1 ventral root).</li>
	<li>Induce a depression in the electrode tank via the syringe by pulling out the piston in order to gently aspirate the nerve rootlet. Then carefully move the electrode to apply it against the spinal cord. 
	Note: Ideally the size of the rootlet matches the size of the electrode opening and thus creates a seal between the internal and external compartments of the electrode. Since differential amplifiers are commonly used for such recordings, any opening between the inside of the electrode and the recording chamber reduces the quality of the signal and makes it difficult to eliminate background noise.</li>
	<li>Start the recording.</li>
	<li>Record the rhythm produced by the preparation under normoxic conditions (i.e., aCSF bubbled with carbogen: 95% O2 and 5% CO2) for at least 20 min to determine the baseline parameters of the preparation.</li>
	<li>Switch the perfusion from carbogen-bubbled aCSF to stimulus aCSF (i.e., bubbled with 95% N2 and 5% CO2 for hypoxia stimulus) for 15 min. Alter exposure duration depending of the experimental protocol. Record the exposure duration on the recording and animal datasheet. Use stainless steel tubes between the aCSF bottle and the chamber whenever possible to avoid gas diffusion to the outside of the tube.</li>
	<li>Switch the perfusion back to standard carbogen-bubbled aCSF for at least 15 min for a recovery recording. Record this on the recording and animal datasheet.</li>
	<li>End the recording.</li>
</ol>
4. Statistical Analysis
<ol>
	<li>From the integrated signal, calculate the frequency as the number of bursts per minute (expressed in bursts/min), the amplitude as the difference between the baseline and the peak of the burst (expressed in mV), the burst duration as the duration from the beginning to the end of the burst (expressed in s), the burst area as the area under the curve of the burst on the integrated signal (expressed in mV&#183;s) (Figure 3).</li>
	<li>Calculate the frequency, amplitude, burst duration and burst area under normoxic (baseline), hypoxic (stimulus) and post-hypoxic (post-stimulus) recovery conditions as the average of the last 5 min of the recordings for each condition. Do not analyze the first portion of each condition because there is a delay between switching the perfusion and aCSF homogenization in the recording chamber.</li>
	<li>Express then stimulus (i.e., hypoxia) and post-stimulus (i.e., post-hypoxic) recovery values as a percentage of the baseline values for the corresponding recording. Express the results as means &#177; S.D.</li>
</ol>

<ol>
	<li>Feldman, J. L., Del Negro, ., A, C., Gray, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+rhythm+of+breathing:+so+near,+yet+so+far.">Understanding the rhythm of breathing: so near, yet so far.</a> Annu Rev Physiol. 75, 423-452 (2013).</li><li>Taylor, A. C., Kollros, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stages+in+the+normal+development+of+Rana+pipiens+larvae.">Stages in the normal development of Rana pipiens larvae.</a> Anat Rec (Hoboken). 94, 7-13 (1946).</li><li>Takeda, R., Remmers, J. E., Baker, J. P., Madden, K. P., Farber, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postsynaptic+potentials+of+bulbar+respiratory+neurons+of+the+turtle.">Postsynaptic potentials of bulbar respiratory neurons of the turtle.</a> Respir Physiol. 64, 149-160 (1986).</li><li>Bouverot, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+breathing+in+birds+compared+with+mammals.">Control of breathing in birds compared with mammals.</a> Physiol Rev. 58, 604-655 (1978).</li><li>Adrian, E. D., Buytendijk, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+changes+in+the+isolated+brain+stem+of+the+goldfish.">Potential changes in the isolated brain stem of the goldfish.</a> J Physiol. 71, 121-135 (1931).</li><li>Suzue, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+rhythm+generation+in+the+in+vitro+brain+stem-spinal+cord+preparation+of+the+neonatal+rat.">Respiratory rhythm generation in the in vitro brain stem-spinal cord preparation of the neonatal rat.</a> J Physiol. 354, 173-183 (1984).</li><li>Fournier, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gestational+stress+promotes+pathological+apneas+and+sex-specific+disruption+of+respiratory+control+development+in+newborn+rat.">Gestational stress promotes pathological apneas and sex-specific disruption of respiratory control development in newborn rat.</a> J Neurosci. 33, 563-573 (2013).</li><li>Caravagna, C., Kinkead, R., Soliz, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-natal+hypoxic+activity+of+the+central+respiratory+command+is+improved+in+transgenic+mice+overexpressing+Epo+in+the+brain.">Post-natal hypoxic activity of the central respiratory command is improved in transgenic mice overexpressing Epo in the brain.</a> Respir Physiol Neurobiol. 200, 64-71 (2014).</li><li>Onimaru, H., Arata, A., Homma, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+mechanisms+of+respiratory+rhythm+generation:+an+approach+using+in+vitro+preparation.">Neuronal mechanisms of respiratory rhythm generation: an approach using in vitro preparation.</a> Jpn J Physiol. 47, 385-403 (1997).</li><li>Onimaru, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+of+the+respiratory+center+using+isolated+brainstem-spinal+cord+preparations.">Studies of the respiratory center using isolated brainstem-spinal cord preparations.</a> Neurosci Res. 21, 183-190 (1995).</li><li>Ballanyi, K., Onimaru, H., Homma, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+network+function+in+the+isolated+brainstem-spinal+cord+of+newborn+rats.">Respiratory network function in the isolated brainstem-spinal cord of newborn rats.</a> Prog Neurobiol. 59, 583-634 (1999).</li><li>Viemari, J. C., Burnet, H., Bevengut, M., Hilaire, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perinatal+maturation+of+the+mouse+respiratory+rhythm-generator:+in+vivo+and+in+vitro+studies.">Perinatal maturation of the mouse respiratory rhythm-generator: in vivo and in vitro studies.</a> Eur J Neurosci. 17, 1233-1244 (2003).</li><li>Rybak, I. A., Abdala, A. P., Markin, S. N., Paton, J. F., Smith, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+organization+and+state-dependent+mechanisms+for+respiratory+rhythm+and+pattern+generation.">Spatial organization and state-dependent mechanisms for respiratory rhythm and pattern generation.</a> Prog Brain Res. , 165-201 (2007).</li><li>Hilaire, G., Viemari, J. C., Coulon, P., Simonneau, M., Bevengut, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+the+respiratory+rhythm+generator+by+the+pontine+noradrenergic+A5+and+A6+groups+in+rodents.">Modulation of the respiratory rhythm generator by the pontine noradrenergic A5 and A6 groups in rodents.</a> Respir Physiol Neurobiol. 143, 187-197 (2004).</li><li>Okada, Y., Kawai, A., Muckenhoff, K., Scheid, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+the+pons+in+hypoxic+respiratory+depression+in+the+neonatal+rat.">Role of the pons in hypoxic respiratory depression in the neonatal rat.</a> Respir Physiol. 111, 55-63 (1998).</li><li>Voituron, N., Frugiere, A., Gros, F., Macron, J. M., Bodineau, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diencephalic+and+mesencephalic+influences+on+ponto-medullary+respiratory+control+in+normoxic+and+hypoxic+conditions:+an+in+vitro+study+on+central+nervous+system+preparations+from+newborn+rat.">Diencephalic and mesencephalic influences on ponto-medullary respiratory control in normoxic and hypoxic conditions: an in vitro study on central nervous system preparations from newborn rat.</a> Neuroscience. 132, 843-854 (2005).</li><li>Somjen, G. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ion+regulation+in+the+brain:+implications+for+pathophysiology.">Ion regulation in the brain: implications for pathophysiology.</a> Neuroscientist. 8, 254-267 (2002).</li><li>Danneman, P. J., Mandrell, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+five+agents/methods+for+anesthesia+of+neonatal+rats.">Evaluation of five agents/methods for anesthesia of neonatal rats.</a> Lab Anim Sci. 47, 386-395 (1997).</li><li>Formenti, A., Zocchi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Error+signals+as+powerful+stimuli+for+the+operant+conditioning-like+process+of+the+fictive+respiratory+output+in+a+brainstem-spinal+cord+preparation+from+rats.">Error signals as powerful stimuli for the operant conditioning-like process of the fictive respiratory output in a brainstem-spinal cord preparation from rats.</a> Behav Brain Res. 272, 8-15 (2014).</li><li>Bierman, A. M., Tankersley, C. G., Wilson, C. G., Chavez-Valdez, R., Gauda, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perinatal+hyperoxic+exposure+reconfigures+the+central+respiratory+network+contributing+to+intolerance+to+anoxia+in+newborn+rat+pups.">Perinatal hyperoxic exposure reconfigures the central respiratory network contributing to intolerance to anoxia in newborn rat pups.</a> J App Physiol. 116, 47-53 (2014).</li><li>Umezawa, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orexin-B+antagonized+respiratory+depression+induced+by+sevoflurane,+propofol,+and+remifentanil+in+isolated+brainstem-spinal+cords+of+neonatal+rats.">Orexin-B antagonized respiratory depression induced by sevoflurane, propofol, and remifentanil in isolated brainstem-spinal cords of neonatal rats.</a> Respir Physiol Neurobiol. 205, 61-65 (2015).</li><li>Ruangkittisakul, A., Secchia, L., Bornes, T. D., Palathinkal, D. M., Ballanyi, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dependence+on+extracellular+Ca2+/K++antagonism+of+inspiratory+centre+rhythms+in+slices+and+en+bloc+preparations+of+newborn+rat+brainstem.">Dependence on extracellular Ca2+/K+ antagonism of inspiratory centre rhythms in slices and en bloc preparations of newborn rat brainstem.</a> J Physiol. 584, 489-508 (2007).</li><li>Cayetanot, F., Bodineau, L., Frugiere, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=5-HT+acting+on+5-HT(1/2)+receptors+does+not+participate+in+the+in+vitro+hypoxic+respiratory+depression.">5-HT acting on 5-HT(1/2) receptors does not participate in the in vitro hypoxic respiratory depression.</a> Neurosci Res. 41, 71-78 (2001).</li><li>Onimaru, H., Homma, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+cell+recordings+from+respiratory+neurons+in+the+medulla+of+brainstem-spinal+cord+preparations+isolated+from+newborn+rats.">Whole cell recordings from respiratory neurons in the medulla of brainstem-spinal cord preparations isolated from newborn rats.</a> Pflugers Archiv : European journal of physiology. 420, 399-406 (1992).</li><li>Paton, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rhythmic+bursting+of+pre-+and+post-inspiratory+neurones+during+central+apnoea+in+mature+mice.">Rhythmic bursting of pre- and post-inspiratory neurones during central apnoea in mature mice.</a> J Physiol. 502 (Pt 3), 623-639 (1997).</li><li>Morin-Surun, M. P., Boudinot, E., Kato, F., Foutz, A. S., Denavit-Saubie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+NMDA+receptors+in+the+respiratory+phase+transition+is+different+in+the+adult+guinea+pig+in+vivo+and+in+the+isolated+brain+stem+preparation.">Involvement of NMDA receptors in the respiratory phase transition is different in the adult guinea pig in vivo and in the isolated brain stem preparation.</a> J Neurophysiol. 74, 770-778 (1995).</li><li>Otsuka, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+volatile+anesthetics+on+respiratory+activity+and+chemosensitivity+in+the+isolated+brainstem-spinal+cord+of+the+newborn+rat.">Effects of volatile anesthetics on respiratory activity and chemosensitivity in the isolated brainstem-spinal cord of the newborn rat.</a> Hokkaido Igaku Zasshi. 73, 117-136 (1998).</li><li>Gestreau, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Task2+potassium+channels+set+central+respiratory+CO2+and+O2+sensitivity.">Task2 potassium channels set central respiratory CO2 and O2 sensitivity.</a> PNAS. 107, 2325-2330 (2010).</li><li>Caravagna, C., Soliz, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PI3K+and+MEK+molecular+pathways+are+involved+in+the+erythropoietin-mediated+regulation+of+the+central+respiratory+command.">PI3K and MEK molecular pathways are involved in the erythropoietin-mediated regulation of the central respiratory command.</a> Respir Physiol Neurobiol. 206C, 36-40 (2014).</li><li>Tree, K., Caravagna, C., Hilaire, G., Peyronnet, J., Cayetanot, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anandamide+centrally+depresses+the+respiratory+rhythm+generator+of+neonatal+mice.">Anandamide centrally depresses the respiratory rhythm generator of neonatal mice.</a> Neuroscience. 170, 1098-1109 (2010).</li><li>Arata, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+activity+of+the+neonatal+dorsolateral+pons+in+vitro.">Respiratory activity of the neonatal dorsolateral pons in vitro.</a> Respir Physiol Neurobiol. 168, 144-152 (2009).</li><li>Onimaru, H., Homma, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+functional+neuron+group+for+respiratory+rhythm+generation+in+the+ventral+medulla.">A novel functional neuron group for respiratory rhythm generation in the ventral medulla.</a> J Neurosci. 23, 1478-1486 (2003).</li><li>St-John, W. M., Paton, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterizations+of+eupnea,+apneusis+and+gasping+in+a+perfused+rat+preparation.">Characterizations of eupnea, apneusis and gasping in a perfused rat preparation.</a> Respir Physiol. 123, 201-213 (2000).</li></ol>

The authors have no competing financial interests or conflicts of interests to disclose.

Accurate quantification of respiratory activity can be challenging. Indeed, breathing is a function that can be both automatic and voluntary, and that is modulated according to the environment, the body&#8217;s needs, the emotional state and the behavior. The advantage of this technique is the isolation of the neural elements responsible for producing the respiratory command. Thus, electrophysiological recordings of brainstem-spinal cord preparations and plethysmography are complementary techniques to study the whole neu...

Breathing is a complex and vital activity controlled by the brain, allowing dioxygen (O2) uptake and carbon dioxide (CO2) elimination. The central respiratory drive is generated by a complex network located in the brainstem in both mammals 1, amphibians 2, reptiles 3, birds 4 and fishes 5. Even if the study of breathing can be processed in vivo, precise mechanistic investigations require direct access to the respiratory control network. To this end, Adrian and Buytendijk developed a reduced goldfish preparation, in which electrodes placed on the brainstem surface record the generated rhythm associated with gill ventilation 5. This approach was subsequently adapted by Suzue in 1984 6 for use in newborn rodents. The advent of this preparation has led to significant advances in respiratory neurobiology. Since it is relatively simple, the technique presented here is amenable to a broad range of basic investigations of rhythmic motor behaviors and their origins in newborn rodents.
The overall goal of this method is to record the neural correlate of inspiratory activity, a respiratory-like rhythm called fictive breathing, produced by the respiratory network. This method can be employed in a wide range of research objectives, targeting inspiratory responses to respiratory variations or pharmacology in both wild type 7 and transgenic 8 animals. Given that experiments are performed at a low temperature, without sensory afferents, and under conditions where the concentrations of glucose and O2 within the aCSF are high, questions have been raised regarding the physiological relevance of the signal recorded. While there are clear differences between in vivo and in vitro conditions (e.g., the frequency of inspiratory bursts) the fact remains that the presence of the core elements of the respiratory network 6 make it possible to study a robust rhythm associated to a vital homeostatic function 9,10.
The rationale behind the development and the use of this technique is to facilitate direct access to the brainstem elements of the respiratory network, which are hardly accessible in vivo, especially in newborns. The brainstem is placed under strictly controlled conditions: the recorded rhythm is not modulated by peripheral afferent inputs from the lungs or the carotid bodies, allowing the study to focus on the central respiratory drive itself 11. Thus, this access is utilized to apply stimuli and record the output signal. In contrast to plethysmography recordings, the respiratory rhythm is modulated by all of its components throughout the body (e.g., lung distension, peripheral chemosensors), making it difficult to apply precise stimuli.
In a newborn rat, the protocol consists of recording the fourth ventral root signal on an isolated brainstem and a truncated spinal cord, maintained in artificial cerebro-spinal fluid (aCSF). The rhythm generated by brainstem-spinal cord preparations is composed of individual slow bursts that are linked to the inspiratory signal 9. Isolated brainstem-spinal cord preparations are easily recordable in rats from post-natal day 0 to 4 (P0 - P4) 7. This approach is commonly used to evaluate the hypoxic response of the respiratory network, and also the response to hypercapnia, acidosis or drugs. An acute hypoxia protocol is presented here. This stimulation is obtained by withdrawal of O2 in the aCSF; this approach is commonly used to assess the tolerance and responsiveness to hypoxic insults. The protocol induces a rhythm depression from the first minute&#160;until the end of the hypoxia exposure (Figure 1)12. This depression is reversed during post-hypoxic recovery 12. Concerning the experimental design, it is important to notice that the pons, located at the rostral part of the brainstem, has an inhibitory action on the rhythm generator 8. Thus, preparations of full brainstem and rostral spinal cord display a lower rhythm. Inclusion of the pons in the isolated sample for the recording is determined according to the goal of the experiment 13; the study of pontine influence on the medulla oblongata network would require recordings with and without the pons to compare the results 14. Moreover, one of the advantages of this technique is the possibility of extending the rostral part of the preparation to include mesencephalic and/or diencephalic regions 15,16, making it possible to assess the effect of these regions on the ponto-medullary respiratory network.

<table><tbody><tr><td>Sylgard</td><td>Sigma Aldrich</td><td>761036-5EA</td><td>Use under hood</td></tr><tr><td>NaCl</td><td>Bioshop</td><td>SOD002</td><td></td></tr><tr><td>KCl</td><td>Bioshop</td><td>POC888</td><td></td></tr><tr><td>CaCl&lt;sub&gt;2&lt;/sub&gt;</td><td>Bioshop</td><td>CCL444</td><td></td></tr><tr><td>MgCl&lt;sub&gt;2&lt;/sub&gt;</td><td>Bioshop</td><td>MAG510</td><td></td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt;</td><td>Bioshop</td><td>SOB999</td><td></td></tr><tr><td>NaH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;</td><td>Bioshop</td><td>SPM306</td><td></td></tr><tr><td>D-glucose</td><td>Bioshop</td><td>GLU501</td><td></td></tr><tr><td>Carbogen</td><td>Linde</td><td>343-02-0006&amp;nbsp;</td><td></td></tr><tr><td>Temperature Controller</td><td>Warner Instruments, Hamden, CT, USA</td><td>TC-324B</td><td></td></tr><tr><td>Suction electrode</td><td>A-M Systems, Everett, WA, USA</td><td>model 573000</td><td></td></tr><tr><td>Differential AC amplifier</td><td>A-M Systems, Everett, WA, USA</td><td>model 1700</td><td></td></tr><tr><td>Moving averager</td><td>CWE, Ardmore, PA, USA</td><td>model MA-821</td><td></td></tr><tr><td>Data acquisition system</td><td>Dataq Instruments, Akron, OH, USA</td><td>model DI-720</td><td></td></tr><tr><td>LabChart software</td><td>ADInstruments, Colorado Springs, CO, USA</td><td></td><td></td></tr><tr><td>Prism sofware</td><td>Graphpad, La Jolla, CA, USA</td><td></td><td></td></tr><tr><td>Dissection chamber</td><td>Plastic box (e.g. petri box) will do</td><td></td><td></td></tr><tr><td>Recording chamber</td><td>Home made</td><td></td><td></td></tr><tr><td>Base</td><td>Kanetec, Bensenville, IL, USA</td><td>MB</td><td></td></tr><tr><td>Micromanipulator</td><td>World Precision Instrument Inc, Sarasota, FL, USA</td><td>KITE-R</td><td></td></tr><tr><td>Base</td><td>Kanetec, Bensenville, IL, USA</td><td>MB</td><td></td></tr><tr><td>Peristaltic pump</td><td>Gilson, Middleton, WI, USA</td><td>MINIPULS 3</td><td></td></tr><tr><td>Faraday Cage</td><td>Home made</td><td></td><td></td></tr><tr><td>Computer</td><td></td><td></td><td></td></tr></tbody></table>

electrophysiology on isolated brainstem-spinal cord preparations from newborn rodents allows neural respiratory network output recording

While it is well known that the central respiratory drive is located in the brainstem, several aspects of its basic function, development, and response to stimuli remain to be fully understood. To overcome the difficulty of accessing the brainstem in the whole animal, isolation of the brainstem and part of the spinal cord is performed. This preparation is maintained in artificial cerebro-spinal fluid where gases, concentrations, and temperature are controlled and monitored. The output signal from the respiratory network is recorded by a suction electrode placed on the fourth ventral root. In this manner, stimuli can be directly applied onto the brainstem, and the effect can be recorded directly. The signal recorded is linked to the inspiratory signal sent to the diaphragm via the phrenic nerve, and can be described as bursts (around 8 bursts per minute). Analysis of these bursts (frequency, amplitude, length, and area under the curve) allows precise characterization of the stimulus effect on the respiratory network. The main limitation of this method is the viability of the preparation beyond the early post-natal stages. Thus, this method greatly focuses on the study of the whole network without the peripheral inputs in the newborn rat.

As mentioned in the introduction, one of the most important advantages of this technique is the direct access to the brainstem to apply various stimuli. As an example, hypoxia was applied here. Figure 1.A.B. displays a full protocol recording, with both normoxic and hypoxic conditions. Figure 1.C.E. displays the rhythm recorded in normoxic conditions (i.e., aCSF bubbled with 95% O2 and 5% CO2 at 26 &#176;C). As previously demonstrated in th...

Watch this Scientific Journal Video about Electrophysiology on Isolated Brainstem-spinal Cord Preparations from Newborn Rodents Allows Neural Respiratory Network Output Recording at JoVE.com

Electrophysiology on Isolated Brainstem-spinal Cord Preparations from Newborn Rodents Allows Neural Respiratory Network Output Recording

The central respiratory drive is located in the brainstem. Spontaneous respiratory motor output from an isolated brainstem-spinal cord is recorded by placing an electrode on the fourth ventral root. This experimental approach is valuable for pharmacological investigations or the assessment of respiratory challenges and genetic manipulations on rhythmic motor behavior.

While it is well known that the central respiratory drive is located in the brainstem, several aspects of its basic function, development, and response to ...

electrophysiology-on-isolated-brainstem-spinal-cord-preparations-from

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

8.5K Views.  Laval University. The central respiratory drive is located in the brainstem. Spontaneous respiratory motor output from an isolated brainstem-spinal cord is recorded by placing an electrode on the fourth ventral root. This experimental approach is valuable for pharmacological investigations or the assessment of respiratory challenges and genetic manipulations on rhythmic motor behavior.

Video: Electrophysiology on Isolated Brainstem-spinal Cord Preparations from Newborn Rodents Allows Neural Respiratory Network Output Recording

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate a method to fabricate suction electrodes that are used to examine CPG activity, or fictive locomotion, in dissected rodent spinal cords. The rodent spinal cords are placed in artificial cerebrospinal fluid and the ventral roots are drawn into the suction electrode. The electrode is constructed by modifying a commercially available suction electrode. A heavier silver wire is used instead of the standard wire given by the commercially available electrode. The glass tip on the commercial electrode is replaced with a plastic tip for increased durability. We prepare hand drawn electrodes and electrodes made from specific sizes of tubing, allowing consistency and reproducibility. Data is collected using an amplifier and neurogram acquisition software. Recordings are performed on an air table within a Faraday cage to prevent mechanical and electrical interference, respectively.

A demonstration of the fabrication and use of an extracellular suction electrode used to measure electrophysiological recordings of neonatal rodent spinal cords in vitro

Development of neural circuitries and locomotion can be studied using neonatal rodent spinal cord central pattern generator (CPG) behavior. We demonstrate ...

Spinal Cord Electrophysiology

The neonatal mouse spinal cord is a model for studying the development of neural circuitries and locomotor movement. We demonstrate the spinal cord dissection and preparation of recording bath artificial cerebrospinal fluid used for locomotor studies. Once dissected, the spinal cord ventral nerve roots can be attached to a recording electrode to record the electrophysiologic signals of the central pattern generating circuitry within the lumbar cord.

A demonstration of the isolation of neonatal mouse spinal cord for electrophysiologic studies.

The neonatal mouse spinal cord is a model for studying the development of neural circuitries and locomotor movement. We demonstrate the spinal cord ...

Biology

Induction of an Isoelectric Brain State to Investigate the Impact of Endogenous Synaptic Activity on Neuronal Excitability In Vivo

The way neurons process information depends both on their intrinsic membrane properties and on the dynamics of the afferent synaptic network. In particular, endogenously-generated network activity, which strongly varies as a function of the state of vigilance, significantly modulates neuronal computation. To investigate how different spontaneous cerebral dynamics impact single neurons&#39; integrative properties, we developed a new experimental strategy in the rat consisting in suppressing in vivo all cerebral activity by means of a systemic injection of a high dose of sodium pentobarbital. Cortical activities, continuously monitored by combined electrocorticogram (ECoG) and intracellular recordings are progressively slowed down, leading to a steady isoelectric profile. This extreme brain state, putting the rat into a deep comatose, was carefully monitored by measuring the physiological constants of the animal throughout the experiments. Intracellular recordings allowed us to characterize and compare the integrative properties of the same neuron embedded into physiologically relevant cortical dynamics, such as those encountered in the sleep-wake cycle, and when the brain was fully silent.

Induction of an Isoelectric Brain State to Investigate the Impact of Endogenous Synaptic Activity on Neuronal Excitability In Vivo

This procedure performs long-lasting in vivo intracellular recordings from single neurons during physiologically relevant cerebral states and after complete abolition of ongoing electrical activities, resulting in an isoelectric brain state. The physiological constants of the animal are carefully monitored during the transition to the artificial comatose condition.

The way neurons process information depends both on their intrinsic membrane properties and on the dynamics of the afferent synaptic network. In ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Preparation of Rhythmically-active In Vitro Neonatal Rodent Brainstem-spinal Cord and Thin Slice

Mammalian inspiratory rhythm is generated from a neuronal network in a region of the medulla called the preB&#246;tzinger complex (pBC), which produces a signal driving the rhythmic contraction of inspiratory muscles. Rhythmic neural activity generated in the pBC and carried to other neuronal pools to drive the musculature of breathing may be studied using various approaches, including en bloc nerve recordings and transverse slice recordings. However, previously published methods have not extensively described the brainstem-spinal cord dissection process in a transparent and reproducible manner for future studies. Here, we present a comprehensive overview of a method used to reproducibly cut rhythmically-active brainstem slices containing the necessary and sufficient neuronal circuitry for generating and transmitting inspiratory drive. This work builds upon previous brainstem-spinal cord electrophysiology protocols to enhance the likelihood of reliably obtaining viable and rhythmically-active slices for recording neuronal output from the pBC, hypoglossal premotor neurons (XII pMN), and hypoglossal motor neurons (XII MN). The work presented expands upon previous published methods by providing detailed, step-by-step illustrations of the dissection, from whole rat pup, to in vitro slice containing the XII rootlets.

This protocol both visually communicates the brainstem-spinal cord preparation and clarifies the preparation of brainstem transverse slices in a comprehensive step-by-step manner. It was designed to increase reproducibility and enhance the likelihood of obtaining viable, long lasting, rhythmically-active slices for recording neural output from the respiratory regions of the brainstem.

Mammalian inspiratory rhythm is generated from a neuronal network in a region of the medulla called the preB&#246;tzinger complex (pBC), which produces a ...

Data Acquisition and Analysis In Brainstem Evoked Response Audiometry In Mice

Brainstem evoked response audiometry (BERA) is of central relevance in the clinical neurophysiology. As other evoked potential (EP) techniques, such as visually evoked potentials (VEPs) or somatosensory evoked potentials (SEPs), the auditory evoked potentials (AEPs) are triggered by the repetitive presentation of identical stimuli, the electroencephalographic (EEG) response of which is subsequently averaged resulting in distinct positive (p) and negative (n) deflections. In humans, both the amplitude and the latency of individual peaks can be used to characterize alterations in synchronization and conduction velocity in the underlying neuronal circuitries. Importantly, AEPs are also applied in basic and preclinical science to identify and characterize the auditory function in pharmacological and genetic animal models. Even more, animal models in combination with pharmacological testing are utilized to investigate for potential benefits in the treatment of sensorineural hearing loss (e.g., age- or noise-induced hearing deficits). Here we provide a detailed and integrative description of how to record auditory brainstem-evoked responses (ABRs) in mice using click and tone-burst application. A specific focus of this protocol is on pre-experimental animal housing, anesthesia, ABR recording, ABR filtering processes, automated wavelet-based amplitude growth function analysis, and latency detection.

Brainstem evoked response audiometry is an important tool in clinical neurophysiology. Nowadays, brainstem evoked response audiometry is also applied in the basic science and preclinical studies involving both pharmacological and genetic animal models. Here we provide a detailed description of how auditory brainstem responses can be successfully recorded and analyzed in mice.

Brainstem evoked response audiometry (BERA) is of central relevance in the clinical neurophysiology. As other evoked potential (EP) techniques, such as ...

In Vivo Intracellular Recording of Type-Identified Rat Spinal Motoneurons During Trans-Spinal Direct Current Stimulation

Intracellular recording of spinal motoneurons in vivo provides a &#8220;gold standard&#8221; for determining the cells&#8217; electrophysiological characteristics in the intact spinal network and holds significant advantages relative to classical in vitro or extracellular recording techniques. An advantage of in vivo intracellular recordings is that this method can be performed on adult animals with a fully mature nervous system, and therefore many observed physiological mechanisms can be translated to practical applications. In this methodological paper, we describe this procedure combined with externally applied constant current stimulation, which mimics polarization processes occurring within spinal neuronal networks. Trans-spinal direct current stimulation (tsDCS) is an innovative method increasingly used as a neuromodulatory intervention in rehabilitation after various neurological injuries as well as in sports. The influence of tsDCS on the nervous system remains poorly understood and the physiological mechanisms behind its actions are largely unknown. The application of the tsDCS simultaneously with intracellular recordings enables us to directly observe changes of motoneuron membrane properties and characteristics of rhythmic firing in response to the polarization of the spinal neuronal network, which is crucial for the understanding of tsDCS actions. Moreover, when the presented protocol includes the identification of the motoneuron with respect to an innervated muscle and its function (flexor versus extensor) as well as the physiological type (fast versus slow) it provides an opportunity to selectively investigate the influence of tsDCS on identified components of spinal circuitry, which seem to be differently affected by polarization. The presented procedure focuses on surgical preparation for intracellular recordings and stimulation with an emphasis on the steps which are necessary to achieve preparation stability and reproducibility of results. The details of the methodology of the anodal or cathodal tsDCS application are discussed while paying attention to practical and safety issues.

This protocol describes in vivo intracellular recording of rat lumbar motoneurons with simultaneous trans-spinal direct current stimulation. The method enables us to measure membrane properties and to record rhythmic firing of motoneurons before, during and after anodal or cathodal polarization of the spinal cord.

Intracellular recording of spinal motoneurons in vivo provides a &#8220;gold standard&#8221; for determining the cells&#8217; electrophysiological ...

Angled Silicon Multielectrode Recordings for Deep Brainstem Structures in Mice

Acute Single-Unit Multi-Electrode Recordings from the Brainstem of Head-Fixed Mice

Silicon multielectrode-based recordings are increasingly popular for studying neuronal activity at the temporal resolution of action potentials in many brain regions. However, recording neuronal activity from deep caudal structures like the brainstem using multi-channel probes remains challenging. A significant concern is finding a trajectory for probe insertion that avoids large blood vessels, such as the superior sagittal venous sinus and the transverse venous sinus. Injuring these large veins can cause extensive bleeding, damage to the underlying brain tissue, and potentially death. This approach describes targeting brainstem structures by coupling anterior coordinates with an angled approach, allowing the recording probe to penetrate the brain below high-risk vascular structures. Compared to a strictly vertical approach, the angled approach maximizes the number of brain regions that can be targeted. Using this strategy, the ventrolateral periaqueductal gray (vlPAG), a brainstem region associated with REM sleep, can be reproducibly and reliably accessed to obtain single-unit, multi-electrode recordings in head-fixed mice before and during sevoflurane anesthesia. The ability to record neuronal activity in the vlPAG and surrounding nuclei with high temporal resolution is a step forward in advancing the understanding of the relationship between REM sleep and anesthesia.

Author Spotlight: Investigating Anesthesia-Induced Sleep Pathways and Neuronal Excitability in Mice

This protocol describes a method to obtain in vivo, high-density single-neuron recordings from the brainstem of head-fixed mice. This approach is deployed to measure the action potential firing of neurons in the ventrolateral periaqueductal gray - a brainstem region inactive during Rapid Eye Movement (REM) sleep - before and during general anesthesia.

Silicon multielectrode-based recordings are increasingly popular for studying neuronal activity at the temporal resolution of action potentials in many ...

Electrophysiology on Isolated Brainstem-spinal Cord Preparations from Newborn Rodents Allows Neural Respiratory Network Output Recording

Summary

Explore More Videos

Chapters in this video

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

Spinal Cord Electrophysiology

Induction of an Isoelectric Brain State to Investigate the Impact of Endogenous Synaptic Activity on Neuronal Excitability In Vivo

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Preparation of Rhythmically-active In Vitro Neonatal Rodent Brainstem-spinal Cord and Thin Slice

Data Acquisition and Analysis In Brainstem Evoked Response Audiometry In Mice

In Vivo Intracellular Recording of Type-Identified Rat Spinal Motoneurons During Trans-Spinal Direct Current Stimulation

Acute Single-Unit Multi-Electrode Recordings from the Brainstem of Head-Fixed Mice