<meta charset="utf8"></head><body>Angled Silicon Multielectrode Recordings for Deep Brainstem Structures in Mice

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<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>저자 스포트라이트: Investigating Anesthesia-Induced Sleep Pathways and Neuronal Excitability in Mice (생쥐의 마취 유도 수면 경로 및 신경 흥분성 조사)

Head-Fixed Mice의 뇌간에서 발생한 급성 단일 단위 다중 전극 기록

Our laboratory investigates the effects of commonly used anesthetics on sleep-related neurons and their communication pathways. We aim to better understand how anesthetics impact endogenous sleep pathways and affect perioperative sleep and cognition. Recently we identified the anatomical substrates for some of the sleep changes caused by certain types of general anesthetics.We did so by using genetically modified mice in which there is endogenous expression of c-Fos, a marker of neuronal activation. However, using this new technique, we will be able to directly measure cellular excitability in groups of neurons in the brainstem that are associated with sleep. This is a major advantage over simply quantifying c-Fos expression as a surrogate marker of neuronal activation.Our next step will be to pursue the cellular and molecular mechanisms underlying the sleep changes induced by certain types of general anesthetics. This will entail measuring cellular excitability in brainstem neurons associated with sleep using neuropixel probes, as well as using spatial transcriptomic approaches to quantify and map the expression of genes related to sleep in these areas.

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

399 조회수.  University of Virginia. 우리 연구실은 일반적으로 사용되는 마취제가 수면 관련 뉴런과 그 전달 경로에 미치는 영향을 조사합니다. 우리는 마취제가 내인성 수면 경로에 어떻게 영향을 미치고 수술 전후 수면 및 인지에 어떤 영향을 미치는지 더 잘 이해하는 것을 목표로 합니다. 최근에 우리는 특정 유형의 전신 마취제에 의해 유발된 수면 변화의 일부에 대한 해부학적 기?...