<meta charset="utf8"></head><body>Registrazioni multielettrodo in silicio angolato per strutture profonde del tronco encefalico nei topi

<ol>
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A., Koch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+origin+of+extracellular+fields+and+currents--EEG,+ECoG,+LFP+and+spikes.">The origin of extracellular fields and currents--EEG, ECoG, LFP and spikes.</a> Nat Rev Neurosci. 13 (6), 407-420 (2012).</li><li>Franklin, K. B. J., Paxinos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The mouse brain in stereotaxic coordinates. , (2008).</li><li>. Pythagorean theorem Available from: <a target="_blank" href="https://en.wikipedia.org/w/index.php?title=Pythagorean_theorem&amp;oldid=1222259845">https://en.wikipedia.org/w/index.php?title=Pythagorean_theorem&amp;oldid=1222259845</a> (2024)</li><li>. The Anatomy of the Laboratory Mouse Available from: <a target="_blank" href="https://www.informatics.jax.org/cookbook/chapters/skeleton.shtml">https://www.informatics.jax.org/cookbook/chapters/skeleton.shtml</a> (2024)</li><li>Cecyn, M. N., Abrahao, K. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Where+do+you+measure+the+Bregma+for+rodent+stereotaxic+surgery.">Where do you measure the Bregma for rodent stereotaxic surgery.</a> IBRO Neurosci Rep. 15, 143-148 (2023).</li><li>Fi&#225;th, R., 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=Slow+insertion+of+silicon+probes+improves+the+quality+of+acute+neuronal+recordings.">Slow insertion of silicon probes improves the quality of acute neuronal recordings.</a> Sci Rep. 9, 111 (2019).</li><li>Wu, J., 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=Transcardiac+perfusion+of+the+mouse+for+brain+tissue+dissection+and+fixation.">Transcardiac perfusion of the mouse for brain tissue dissection and fixation.</a> Bio-protocol. 11 (5), e3988 (2021).</li><li>Peters, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> petersaj/AP_histology. , (2024).</li><li>Pachitariu, M., Sridhar, S., Stringer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Solving the spike sorting problem with Kilosort. , (2023).</li><li>Akam, T., Walton, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=pyPhotometry:+Open+source+Python+based+hardware+and+software+for+fiber+photometry+data+acquisition.">pyPhotometry: Open source Python based hardware and software for fiber photometry data acquisition.</a> Sci Rep. 9 (1), 3521 (2019).</li><li>Leiras, R., Cregg, J. M., Kiehn, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brainstem+circuits+for+locomotion.">Brainstem circuits for locomotion.</a> Ann Rev Neurosci. 45 (2022), 63-85 (2022).</li><li>Benarroch, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brainstem+integration+of+arousal,+sleep,+cardiovascular,+and+respiratory+control.">Brainstem integration of arousal, sleep, cardiovascular, and respiratory control.</a> Neurology. 91 (21), 958-966 (2018).</li><li>Guyenet, P. G., Bayliss, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+control+of+breathing+and+CO2+homeostasis.">Neural control of breathing and CO2 homeostasis.</a> Neuron. 87 (5), 946-961 (2015).</li><li>Potter, K. A., Buck, A. C., Self, W. K., Capadona, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stab+injury+and+device+implantation+within+the+brain+results+in+inversely+multiphasic+neuroinflammatory+and+neurodegenerative+responses.">Stab injury and device implantation within the brain results in inversely multiphasic neuroinflammatory and neurodegenerative responses.</a> J Neural Eng. 9 (4), 046020 (2012).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1024 channel RHD Recording Controller</td>
			<td>Intan Technologies, Los Angeles, California, USA</td>
			<td>C3008</td>
			<td>Silicon probe recording; recording hardware and software</td>
			<td></td>
		</tr>
		<tr>
			<td>24 mm x 50 mm No. 1.5 VWR coverslip</td>
			<td>VWR, Radnor, Pennsylvania, USA</td>
			<td>48393-081</td>
			<td>Histology</td>
			<td></td>
		</tr>
		<tr>
			<td>4% PFA in PBS</td>
			<td>ThermoFisher Scientific, Waltham, Massachusetts, USA</td>
			<td>J61899.AK</td>
			<td>Histology; perfusion solution</td>
			<td></td>
		</tr>
		<tr>
			<td>C&#38;B metabond</td>
			<td>Patterson Dental, Richmond, Virginia, USA</td>
			<td>powder: 5533559, quick base: 5533492, catalyst: 55335007</td>
			<td>Headplate &#38;Headset Implantation</td>
			<td></td>
		</tr>
		<tr>
			<td>C57/6J mice 4-6 weeks, males</td>
			<td>The Jackson Laboratory, Bar Harbor, Maine, USA</td>
			<td>000664</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Capnomac Ultima</td>
			<td>Datex, Helsinki, Finland&#160;</td>
			<td>ULT-SVi-27-07</td>
			<td>Gas Analyzer; discontinued; alternative gas analyzer can be purchased from Bionet America&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>CM-DiI</td>
			<td>ThermoFisher Scientific, Waltham, Massachusetts, USA</td>
			<td>V22888</td>
			<td>Red fluorescent dye for coating of the silicon probe</td>
			<td></td>
		</tr>
		<tr>
			<td>Connector Header</td>
			<td>DigiKey, Thief River Falls, Minnesota, USA</td>
			<td>1212-1788-ND</td>
			<td>ECoG Headset</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI Fluoromount-G</td>
			<td>SouthernBiotech, Birmingham, Alabama, USA</td>
			<td>0100-20</td>
			<td>Histology</td>
			<td></td>
		</tr>
		<tr>
			<td>iBOND Universal</td>
			<td>Patterson Dental, Richmond, Virginia, USA</td>
			<td>044-1113</td>
			<td>Headplate &#38;Headset Implantation; for&#160; securing stainless steel wires to the skull</td>
			<td></td>
		</tr>
		<tr>
			<td>Low toxicity silicon adhesive</td>
			<td>World Precision Instruments, Sarasota, Florida, USA</td>
			<td>KWIK-SIL</td>
			<td>Headplate</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-Manipulator System</td>
			<td>New Scale Technologies, Victor, New York, USA</td>
			<td>Multi-Probe Manipulator: XYZ Stage Assembly: 06464-0000, MPM System Kit: 06267-3-0001, MPM-Platform-360, MPM ring for MPM Manual Arms, MPM_Ring-72 DEG: 06262-3-0000</td>
			<td>Silicon probe recording; inserting the probe into the brain</td>
			<td></td>
		</tr>
		<tr>
			<td>Microprobes</td>
			<td>UCLA, Los Angeles, California, USA</td>
			<td>256 ANS, 64M</td>
			<td>Discontinued; alternative silicon probes can be purchased from Neuropixels</td>
			<td></td>
		</tr>
		<tr>
			<td>Mineral Oil</td>
			<td>Sigma Aldrich, Saint Luis, Missouri, USA</td>
			<td>M8410-100ML</td>
			<td>Silicon probe recording; preventing the tissue from drying during the recording</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal saline</td>
			<td>ThermoFisher Scientific, Waltham, Massachusetts, USA</td>
			<td>Z1376</td>
			<td>Headplate &#38;Headset Implantation; preventing the brain from drying during the surgery</td>
			<td></td>
		</tr>
		<tr>
			<td>PFA-Coated Stainless Steel Wire-Diameter 0.008 in. coated with striped ends</td>
			<td>A-M systems, Sequim, Washington, USA</td>
			<td>791400</td>
			<td>ECoG Headset &#38; reference electrode for ECoG&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Platinum wire 24AWG&#160;</td>
			<td>World Precision Instruments, Sarasota, Florida, USA</td>
			<td>PTP201</td>
			<td>Reference electrode for the silicon probe recording&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Shandon Colorfrost Plus microscope slides</td>
			<td>ThermoFisher Scientific, Waltham, Massachusetts, USA</td>
			<td>99-910-01</td>
			<td>Histology</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel Headplate</td>
			<td>Star Rapid, China</td>
			<td>custom made part</td>
			<td>Headplate &#38;Headset Implantation; design available upon request</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic apparatus</td>
			<td>KOPF, Tujunga, California, USA</td>
			<td>Model 940 Small Animal Stereotaxic Instrument with Digital Display Console</td>
			<td>Headplate &#38;Headset Implantation</td>
			<td></td>
		</tr>
	</tbody>
</table>

acute single-unit multi-electrode recordings from the brainstem of head-fixed mice

Silicon multielectrode-based recordings are becoming increasingly popular to measure neuronal activity across many brain regions with single action potential resolution1,2,3,4. Over the last decade, high-density recording technology has grown considerably. Current silicon-based recording electrodes can accommodate high channel counts, optical fibers, and electrocorticography (ECoG) recording devices5,6. Moreover, chronic implantation of these electrodes allows for long-term recordings7,8.
Despite recent technological advancements, targeting deep caudal structures like the brainstem with multi-channel probes remains challenging. When targeting brainstem structures such as the ventrolateral periacqueductal gray (vlPAG), one significant obstacle is identifying a probe trajectory that avoids major blood vessels, e.g., the superior sagittal venous sinus and the transverse venous sinus. Injury to these large veins can cause extensive bleeding, damage to the underlying brain tissue, and even death9,10. We propose targeting brainstem structures from anterior coordinates at an angle, allowing the recording probe to penetrate the brain below such high-risk vascular structures (see Figure 1). This angled approach, compared to a vertical one, maximizes the number of brain regions accessible for recording. Additionally, in experimental circumstances wherein ECoG recordings are desired, the angled anterior approach affords more skull surface available for ECoG headset implantation, as the craniotomy window for probe insertion is positioned more anteriorly10,11.
Identifying the specific cell groups and circuits responsible for anesthesia-induced REM sleep changes remains a major goal of anesthesia research. Thus, the objective here was to reproducibly and reliably access the vlPAG - a brainstem region associated with REM sleep - to obtain single unit, multi-electrode recordings in head-fixed mice before and during sevoflurane anesthesia12,13. Previous studies have used electrophysiological local field potential (LFP) measurements of the vlPAG in awake mice to identify neural state changes associated with anesthesia14,15. However, LFP measurements are primarily sensitive to synaptic activity, not action potentials, within the recorded area16. Consequently, there remains a limited understanding of how anesthetics directly affect the neural activity patterns produced by vlPAG neurons. Here, a method is described to obtain high-density single-neuron recordings from the brainstem of head-fixed mice. This method can also be adapted to record single-neuron activity from various other deep and posterior brainstem structures.

<meta charset="utf8"></head><body>Autore in primo piano: Studio delle vie del sonno indotte dall'anestesia e dell'eccitabilità neuronale nei topi

Registrazioni acute multi-elettrodo a unità singola dal tronco encefalico di topi fissati alla testa

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

acute-single-unit-multi-electrode-recordings-from-brainstem-head

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Research

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

JoVE Journal

Neuroscience

437 Views.  University of Virginia. Il nostro laboratorio studia gli effetti degli anestetici comunemente usati sui neuroni legati al sonno e sulle loro vie di comunicazione. Il nostro obiettivo è quello di comprendere meglio come gli anestetici influiscono sui percorsi endogeni del sonno e influenzano il sonno perioperatorio e la cognizione. Recentemente abbiamo identificato i substrati anatomici per alcuni dei cambiamenti del sonno causati da alcuni tipi di anestetici generali.Lo abbi...

Video: Autore in primo piano: Studio delle vie del sonno indotte dall'anestesia e dell'eccitabilità neuronale nei topi

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.

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.