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במאמר זה SEEG שימש כדי לחקור את הפעילות של אוכלוסיות נוירונים המקומיות בdACC במהלך משימת קבלת החלטות בבני אדם. העבודה קודמת חקרה את הפעילות של נוירונים בודדים בdACC באמצעות הקלטות microelectode במהלך ניתוח 14 והוכיחה כי פעילות dACC היא מווסתת על ידי פעילות קודמת. מחקרי microelectro...

אפילפסיה, הפרעת נוירולוגית שכיחה המתאפיינת בהתקפים חוזרים ונשנים מרובים לאורך זמן, מהווה 1% מהנטל העולמי של מחלות 1. תרופות נגד אפילפסיה לא מצליחות לשלוט בהתקפים ב -20 - 30% מחולי 2,3. בחולים רפואיים סוררים הללו, ניתוח אפילפסיה מצויינים לעתים קרובות 4,5. ההחלטה להמשיך עם ניתוח דורשת איתור מוקד התפיסה, תנאי מוקדם לגיבוש תכנית כירורגית. בתחילה, טכניקות לא פולשנית משמשות לlateralize ולמקם מוקד התפיסה. Electroencephalography (EEG), למשל, פעילות חשמלית בקליפת המוח אמצעים שנרשם מאלקטרודות המונחות על הקרקפת, ולעתים קרובות יכולה לספק מספיק מידע על מיקומו של מוקד התפיסה. בנוסף, הדמיה בתהודה מגנטית (MRI) יכולה להוכיח נגעים בדידים, כגון טרשת בהיפוקמפוס, פתולוגיה הקלאסית ראתה בצורה הנפוצה ביותר של אפילפסיה רפואית סוררת, לא mesialאפילפסיה emporal אונה (MTLE). לעתים קרובות, עם זאת, הבירור לא פולשנית אינו יכול לזהות מוקד תפיסה. במקרים אלה, electrocorticography פולשנית (ECOG) עם אלקטרודות תוך-מוחיות נדרש למקם את המיקוד ולהנחות את טיפול כירורגים נוסף 6. ECOG הוא טכניקת neurophysiological המשמשת למדידת פעילות חשמלית באמצעות אלקטרודות המונחות במגע ישיר עם המוח. רשתות או רצועות של משטח אלקטרודות (subdural) ממוקמות על פני השטח של המוח, תהליך שדורש פתיחת גולגולת (הסרת דש עצם) ופתיחה גדולה של הדורה. ניתן להציב אלקטרודות משטח אלה מעל האזור המשוער (ים) של תחילת התקף. הקצוות המרוחקים של האלקטרודות מנהרה דרך פתחים קטנים בעור ומחובר לציוד ההקלטה ביחידת ניטור אפילפסיה (EMU). בEMU, החולה נתון למעקב לפעילות התקפים קליני באמצעות וידאו רציף והקלטות ECOG. i טכניקה זושימושי לאיסוף לטווח ארוך (ימים עד שבועות) הקלטות של ictal והתפרקויות חשמליות interictal על פני שטחים גדולים יחסית של פני השטח של קליפת המוח של. בעוד הקלטות תוך גולגולתי אלה לא יסולא בפז קליני לחקירת מוקדים של התקפים והתפשטות, הם גם מספקים לנו את ההזדמנות כדי לחקור את תפקוד הקוגניטיבי ונוירופיזיולוגיה בבני אדם עוברים משימות התנהגותיות שתוכננו במיוחד. ECOG באמצעות אלקטרודות רשת subdural נעשה שימוש כדי לחקור היבטים שונים של תפקוד קליפת המוח, לרבות עיבוד חושי ושפה. כדוגמה אחת מיני רבות, Bouchard et al הראה התיאום הזמני של שרירי הפה בהיווצרותה של הברות לשפה מדוברת בקליפת המוח הסנסורית הגחון, אזור שזוהה כקליפה הסנסורית הדיבור האנושי 7. יתר על כן, ECOG עם מיקום רשת subdural גם נוצל כדי לחקור את המנגנונים שבאמצעותם בני האדם מסוגלים attenד לקול מסוים בתוך קהל: מה שנקרא &quot;אפקט מסיבת הקוקטייל &#39;8,9. הקלטות ECOG הוכיחו כי יש שתי להקות שונות עצביות שאופן דינמי לעקוב אחר זרמי דיבור, תנודות שני שלב בתדר נמוך וגבוה גמא האמפליטודה, וכי יש אתרי עיבוד שונים - אתר אחד &#39;אפנון&#39; העוקב אחר שני הרמקולים, ו&#39;בחירה &#39;אחד אתר העוקב אחר דברן השתתף 5. יישום אחר המתעוררים מECOG עם מיקום האלקטרודה subdural הוא הפוטנציאל לשימוש עם מוח ממשקי מחשב (BCIs), שבו &quot;לפענח&quot; פעילות עצבית כדי לנהוג פלט חיצוני. טכנולוגיה זו יש את הפוטנציאל של מתן חולים עם מוח חמור או פגיעות בעמוד השדרה כדי לתקשר עם העולם ולטפל תותבות 10,11. בעוד מיקום רשת subdural תרם רב להבנת העל שלנואזורים בקליפת המוח ficial ושימושי בזיהוי מוקדי epileptogenic קליפת המוח, טכניקה זו דורשת פתיחת גולגולת וסיכונים הכרוכים בו, והוא מוגבל בדרך כלל ללימוד על פני השטח החיצוניים של המוח. electroencephalography stereotactic (SEEG) הוא טכניקה המאפשרת ההערכה של מוקדי epileptogenic עמוקים 12. עם היסטוריה של שימוש בצרפת ובאיטליה ארוכה, הוא גם יותר ויותר בשימוש בארה&quot;ב 13. SEEG כרוך המיקום של אלקטרודות מרובות (בדרך כלל 10-16) עמוק בתוך החומר של המוח באמצעות (כמה מ&quot;מ) חורים בר תרגיל טוויסט קטנים. יתרונות של SEEG על מיקום רשת subdural כוללים טבעה פחות פולשניות, את הקלות של בחינת ההמיספרות דו-צדדיות בעת צורך, ואת היכולת ליצור מפות תלת-ממדיות של התפשטות התקף. יתר על כן, אלקטרודות אלה מאפשרים זיהוי של מוקדי epileptogenic העמוקים שהיו קשים בעבר להזדהות עם אלקטרודות פני השטח. הליך זה גם מעמיד לes ההזדמנות לחקור נוירופיזיולוגיה ותפקוד של מבנים בקליפת המוח עמוקים, כגון מערכת הלימבית, קליפת mesoparietal, קליפת mesotemporal, וקליפת orbitofrontal, שכולן היו קשה בעבר לחקור ישירות בבני אדם. מסמך זה מדגים כיצד ניתן לנצל SEEG לחקור את תפקוד הקוגניטיבי בקליפת המוח הקדמית גב cingulate (dACC). DACC הוא אזור במוח שנחקר באופן נרחב, אבל זה גם אחד מהבין הכי גרוע. נחשב אזור משמעותי להכרה אנושית, סביר להניח כי dACC הוא מרכזי לעיבוד העצבי הדינמי של החלטות בהקשר של דרישות משתנות ללא הרף שהוטלו על ידי הסביבה 14. מחקרים בשני הקופים ובני האדם 15,16 17 מצביעים על כך שdACC משלב סיכונים והטבות פוטנציאליים של פעולה מסוימת, במיוחד במצבים של סותר בו זמנית דורש 18-21, ומטרodulates החלטות אלה בהקשר של פעולות קודמות והתוצאות שלהם 14,22,23. התערבות רב-מקור המשימות (MSIT), משימה התנהגותיות כמו שטרופ, משמשת לעתים קרובות כדי לחקור עיבוד סכסוך בdACC. משימת MSIT מפעילה את dACC על ידי גיוס נוירונים מעורבים במספר תחומים של עיבוד מוסדרים על ידי dACC 24,25. משימה זו באופן ספציפי מפעילה את dACC על ידי בדיקת תכונות של קבלת החלטות, איתור היעד, זיהוי חידוש, זיהוי שגיאות, בחירת תגובה, ותחרות גירוי / תגובה. בנוסף, משימת MSIT מציגה ממדים רבים של הפרעות קוגניטיביות, אשר מנוצלות במחקר זה כדי לחקור תגובות עצביות dACC לגירויים סותרים בו זמנית באמצעות SEEG.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Trigger I/O cable</td>
			<td>Natus Medical Inc.</td>
			<td>5029</td>
			<td>PS2 to BNC cable</td>
		</tr>
		<tr>
			<td>BNC cables for analog pulses</td>
			<td></td>
			<td></td>
			<td>Can be ordered from most electronics stores.</td>
		</tr>
		<tr>
			<td>Power strip with surge protection and battery backup</td>
			<td>Tripp Lite</td>
			<td>SMART500RT1U UPC</td>
			<td>Power source and backup</td>
		</tr>
		<tr>
			<td>National instruments multifunctional daq data acquisition box NI PCIe-6382 DAQ cards</td>
			<td>National Instruments</td>
			<td>PCIe-6382 w/ BNC 2090A</td>
			<td>PCI cards for behavioral control interface</td>
		</tr>
		<tr>
			<td>Custom made button box - human interface device</td>
			<td></td>
			<td></td>
			<td>Any human interface device with three buttons may be used. Alternatively, 3 keyboard buttons may be used.</td>
		</tr>
		<tr>
			<td>Xltek 128 channel clinical intracranial EEG monitoring system EMU128FS</td>
			<td>Natus Medical Inc.</td>
			<td>002047c</td>
			<td>Clinical recording system</td>
		</tr>
		<tr>
			<td>Subject monitor and associated cables for visual stimulus presentation</td>
			<td>Dell</td>
			<td>U2212HMc</td>
			<td>Most Monitors are adequate here.</td>
		</tr>
		<tr>
			<td>Personal comptuer running behavioral software with DAQ cards installed</td>
			<td>Superlogics</td>
			<td>SL-2U-PD-Q87SLQ-BA</td>
			<td>Computer for recording neural data</td>
		</tr>
		<tr>
			<td>Mains cable for monitor</td>
			<td></td>
			<td></td>
			<td>Usually comes with the monitor, can be purchased at any electronics store.</td>
		</tr>
		<tr>
			<td>Monkey Logic software which runs on Matlab 2010A</td>
			<td></td>
			<td></td>
			<td>Free from MonkeyLogic website</td>
		</tr>
		<tr>
			<td>MATLAB 2010a software with data acquisition toolbox</td>
			<td>Mathworks</td>
			<td></td>
			<td>Matlab software</td>
		</tr>
		<tr>
			<td>sEEG electrodes AD TECH or PMT</td>
			<td>AD TECH</td>
			<td>2102-##-101</td>
			<td>Platinum tip, diameter (0.89 mm, 1 mm, 1.1 mm), uninsulated length 2.3 mm; The ## in the catalog number indicates the number of contacts on the electrode (08, 10, 12, or 16)</td>
		</tr>
		<tr>
			<td>Cabrio connectors</td>
			<td>PMT</td>
			<td>2125-##-01</td>
			<td>The ## in the catalog number indicates the number of contacts on the electrode (08, 10, 12, or 16)</td>
		</tr>
		<tr>
			<td>Tucker Davis Technologies Amplifier</td>
			<td>Tucker Davs Technologies</td>
			<td>PZ5</td>
			<td>preamplifier for neural data</td>
		</tr>
		<tr>
			<td>Tucker Davis Technologies processor</td>
			<td>Tucker Davs Technologies</td>
			<td>RZ2</td>
			<td>Neural signal processor for neural data</td>
		</tr>
		<tr>
			<td>TuckerDavis Technologies data streamer</td>
			<td>Tucker Davs Technologies</td>
			<td>RS4</td>
			<td>Data streamer and storage</td>
		</tr>
		<tr>
			<td>Fiber optics cables to connect TDT systems</td>
			<td>Tucker Davs Technologies</td>
			<td>F05</td>
			<td>Fiber optic cables for connecting Tucker Davis Technologies&#39; prodcuts.</td>
		</tr>
		<tr>
			<td>ribbon cable and snap serial connector for digital markers</td>
			<td></td>
			<td></td>
			<td>Can be ordered from ost electronics stores.</td>
		</tr>
		<tr>
			<td>personal computer fro running TDT RPvdsEx and OpenEx software</td>
			<td>Superlogics</td>
			<td>SL-2U-PD-Q87SLQ-BA</td>
			<td>computer for behavioral control</td>
		</tr>
		<tr>
			<td>middle atlantics server cabinet with casters</td>
			<td>Middle Atlantic Products</td>
			<td>PTRK-21</td>
			<td>Server case to house all of the research items</td>
		</tr>
		<tr>
			<td>Tucker Davis Technologies splitter box to split clinical and research recrodings</td>
			<td>Tucker Davs Technologies</td>
			<td></td>
			<td>This splitter box is a semi-custom device. Researchers should consult the attending neurologists about splitting the research and clinical recordings in a way that doesn&#39;t interfere with clinical care.</td>
		</tr>
		<tr>
			<td>Researcher monitor with requisite cables</td>
			<td>Dell</td>
			<td>U2212HMc</td>
			<td>Most Monitors are adequate here.</td>
		</tr>
		<tr>
			<td>button box power source - 5 volts, 2 amperes</td>
			<td></td>
			<td></td>
			<td>Can be purchased at any electronics store.</td>
		</tr>
		<tr>
			<td>TDT optical interface PCI card</td>
			<td>Tucker Davs Technologies</td>
			<td>P05</td>
			<td></td>
		</tr>
	</tbody>
</table>

investigating the function of deep cortical and subcortical structures using stereotactic electroencephalography: lessons from the anterior cingulate cortex

Stereotactic Electroencephalography (SEEG) הוא טכניקה המשמשת למקם מוקדים של התקפים בחולים עם אפילפסיה רפואית סוררת. הליך זה כרוך המיקום הכרוני של אלקטרודות עומק מרובה לתוך האזורים של המוח בדרך כלל אינו נגיש באמצעות מיקום האלקטרודה רשת subdural. SEEG כך מספקת הזדמנות ייחודית לחקור את תפקוד מוח. במאמר זה אנו מדגימים כיצד SEEG יכול לשמש כדי לחקור את תפקידו של רכס החגורה קדמית הגבי (dACC) בשליטה קוגניטיבית. אנו כוללים תיאור של הליך SEEG, הממחיש את המיקום כירורגית של אלקטרודות. אנו מתארים את המרכיבים ותהליך הנדרשים כדי להקליט פוטנציאל שדה מקומי נתונים (LFP) מההסכמה נבדקת בעת שעוסקות במשימה התנהגותית. בדוגמא בתנאי, נושאים לשחק משימת הפרעות קוגניטיביות, ואנו מדגימים כיצד אותות נרשמים ונותחו מאלקטרודות ברכס החגורה הקדמית גב, intim אזורמעורב המצוי בתהליך קבלת החלטות. נסיים בהצעות נוספות של דרכים שבהן שיטה זו יכולה לשמש לחקירת תהליכים קוגניטיביים אנושיים.

ברגע שמטופל נבחר למיקום האלקטרודה SEEG, הוא / היא עוברת ניגוד T2 וT1 נפח המשופר MRI. לאחר מכן מסלולי אלקטרודה SEEG מתוכננים באמצעות ניווט stereotactic של רצפי MRI נפח (איור 1). טכניקה זו מאפשרת לאוסף של פוטנציאל בתחום מקומי ממבנים עמוקים בתוך קליפת המוח כגון קליפת מוח קדמי גב ...

Watch this Scientific Journal Video about Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex a

Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex

Stereotactic Electroencephalography (SEEG) הוא טכניקה ניתוחית המשמשת בניתוחי אפילפסיה לעזור למקם מוקדים של התקפים. זה גם מאפשר הזדמנות ייחודית לחקור את תפקוד מוח. כאן אנו מתארים כיצד SEEG יכול לשמש כדי לחקור תהליכים קוגניטיביים בבני אדם.

חוקרים את הפונקציה של עומק קליפת מוח ומבנים קורטיקליים שימוש stereotactic Electroencephalography: לקחים מCortex Anterior cingulate

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure ...

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Neuroscience

12.1K Views.  Columbia University Medical Center, New York Presbyterian Hospital.  Stereotactic Electroencephalography (SEEG) הוא טכניקה ניתוחית המשמשת בניתוחי אפילפסיה לעזור למקם מוקדים של התקפים. זה גם מאפשר הזדמנות ייחודית לחקור את תפקוד מוח. כאן אנו מתארים כיצד SEEG יכול לשמש כדי לחקור תהליכים קוגניטיביים בבני אדם. 

Video: Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex

Investigating Social Cognition in Infants and Adults Using Dense Array Electroencephalography (dEEG)

Dense array electroencephalography (dEEG), which provides a non-invasive window for measuring brain activity and a temporal resolution unsurpassed by any other current brain imaging technology1,2, is being used increasingly in the study of social cognitive functioning in infants and adults. While dEEG is enabling researchers to examine brain activity patterns with unprecedented levels of sensitivity, conventional EEG recording systems continue to face certain limitations, including 1) poor spatial resolution and source localization3,4,2) the physical discomfort for test subjects of enduring the individual application of numerous electrodes to the surface of the scalp, and 3) the complexity for researchers of learning to use multiple software packages to collect and process data. Here we present an overview of an established methodology that represents a significant improvement on conventional methodologies for studying EEG in infants and adults. Although several analytical software techniques can be used to establish indirect indices of source localization to improve the spatial resolution of dEEG, the HydroCel Geodesic Sensor Net (HCGSN) by Electrical Geodesics, Inc. (EGI), a dense sensory array that maintains equal distances among adjacent recording electrodes on all surfaces of the scalp, further enhances spatial resolution4,5,6 compared to standard dEEG systems. The sponge-based HCGSN can be applied rapidly and without scalp abrasion, making it ideal for use with adults7,8, children9,10,11, and infants12, in both research and clinical4,5,6,13,14,15 settings. This feature allows for considerable cost and time savings by decreasing the average net application time compared to other dEEG systems. Moreover, the HCGSN includes unified, seamless software applications for all phases of data, greatly simplifying the collection, processing, and analysis of dEEG data.
The HCGSN features a low-profile electrode pedestal, which, when filled with electrolyte solution, creates a sealed microenvironment and an electrode-scalp interface. In all Geodesic dEEG systems, EEG sensors detect changes in voltage originating from the participant's scalp, along with a small amount of electrical noise originating from the room environment. Electrical signals from all sensors of the Geodesic sensor net are received simultaneously by the amplifier, where they are automatically processed, packaged, and sent to the data-acquisition computer (DAC). Once received by the DAC, scalp electrical activity can be isolated from artifacts for analysis using the filtering and artifact detection tools included in the EGI software. Typically, the HCGSN can be used continuously for only up to two hours because the electrolyte solution dries out over time, gradually decreasing the quality of the scalp-electrode interface.
In the Parent-Infant Research Lab at the University of Toronto, we are using dEEG to study social cognitive processes including memory, emotion, goals, intentionality, anticipation, and executive functioning in both adult and infant participants.

Investigating Social Cognition in Infants and Adults Using Dense Array Electroencephalography dEEG

Dense array electroencephalography is being used increasingly to study social cognitive functions in infants and adults. Here we present an established methodology that represents a significant improvement on conventional methodologies for studying EEG in infants and adults.

חקירת קוגניציה חברתית תינוקות ומבוגרים שימוש Electroencephalography מערך דחוס ( ד EEG)

Dense array electroencephalography (dEEG), which provides a non-invasive window for measuring brain activity and a temporal resolution unsurpassed by any ...

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

Electroencephalography סימולטני, מדידה בזמן אמת של הריכוז לקטט ומניפולצית Optogenetic של פעילות עצבית בקליפת המוח המכרסמת

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

Juxtasomal Biocytin Labeling to Study the Structure-function Relationship of Individual Cortical Neurons

The cerebral cortex is characterized by multiple layers and many distinct cell-types that together as a network are responsible for many higher cognitive functions including decision making, sensory-guided behavior or memory. To understand how such intricate neuronal networks perform such tasks, a crucial step is to determine the function (or electrical activity) of individual cell types within the network, preferentially when the animal is performing a relevant cognitive task. Additionally, it is equally important to determine the anatomical structure of the network and the morphological architecture of the individual neurons to allow reverse engineering the cortical network. Technical breakthroughs available today allow recording cellular activity in awake, behaving animals with the valuable option of post&#160;hoc identifying the recorded neurons. Here, we demonstrate the juxtasomal biocytin labeling technique, which involves recording action potential spiking in the extracellular (or loose-patch) configuration using conventional patch pipettes. The juxtasomal recording configuration is relatively stable and applicable across behavioral conditions, including anesthetized, sedated, awake head-fixed, and even in the freely moving animal. Thus, this method allows linking cell-type specific action potential spiking during animal behavior to reconstruction of the individual neurons and ultimately, the entire cortical microcircuit. In this video manuscript, we show how individual neurons in the juxtasomal configuration can be labeled with biocytin in the urethane-anaesthetized rat for post&#160;hoc identification and morphological reconstruction.

To understand the structure of neuronal networks, functional and morphological characterization of individual neurons is a necessity. Here, we demonstrate juxtasomal biocytin labeling, which allows electrophysiological recordings in the extracellular configuration, yet maintaining the ability to intracellularly label the neuron for post&#160;hoc reconstruction of dendritic and axonal architecture.

Juxtasomal Biocytin תיוג לחקר קשר בין מבנה לתפקוד של פרט נוירונים קליפתיים

The cerebral cortex is characterized by multiple layers and many distinct cell-types that together as a network are responsible for many higher cognitive ...

Analysis of Gene Expression Changes in the Rat Hippocampus After Deep Brain Stimulation of the Anterior Thalamic Nucleus

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective treatment for several movement disorders that have failed to respond to medication. Recent progress in the field of DBS surgery has begun to extend the application of this surgical technique to other conditions as diverse as morbid obesity, depression and obsessive compulsive disorder. Despite these expanding indications, little is known about the underlying physiological mechanisms that facilitate the beneficial effects of DBS surgery. One approach to this question is to perform gene expression analysis in neurons that receive the electrical stimulation. Previous studies have shown that neurogenesis in the rat dentate gyrus is elicited in DBS targeting of the anterior nucleus of the thalamus1. DBS surgery targeting the ATN is used widely for treatment refractory epilepsy. It is thus of much interest for us to explore the transcriptional changes induced by electrically stimulating the ATN. In this manuscript, we describe our methodologies for stereotactically-guided DBS surgery targeting the ATN in adult male Wistar rats. We also discuss the subsequent steps for tissue dissection, RNA isolation, cDNA preparation and quantitative RT-PCR for measuring gene expression changes. This method could be applied and modified for stimulating the basal ganglia and other regions of the brain commonly clinically targeted. The gene expression study described here assumes a candidate target gene approach for discovering molecular players that could be directing the mechanism for DBS.

The mechanism underlying the therapeutic effects of Deep Brain Stimulation (DBS) surgery needs investigation. The methods presented in this manuscript describe an experimental approach to examine the cellular events triggered by DBS by analyzing the gene expression profile of candidate genes that can facilitate neurogenesis post DBS surgery.

ניתוח שינויי ביטוי הגנים בהיפוקמפוס העכברוש לאחר Deep Brain Stimulation של Anterior התלמוס הגרעין

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective ...

Imaging Serotonergic Fibers in the Mouse Spinal Cord Using the CLARITY/CUBIC Technique

Long descending fibers to the spinal cord are essential for locomotion, pain perception, and other behaviors. The fiber termination pattern in the spinal cord of the majority of these fiber systems have not been thoroughly investigated in any species. Serotonergic fibers, which project to the spinal cord, have been studied in rats and opossums on histological sections and their functional significance has been deduced based on their fiber termination pattern in the spinal cord. With the development of CLARITY and CUBIC techniques, it is possible to investigate this fiber system and its distribution in the spinal cord, which is likely to reveal previously unknown features of serotonergic supraspinal pathways. Here, we provide a detailed protocol for imaging the serotonergic fibers in the mouse spinal cord using the combined CLARITY and CUBIC techniques. The method involves perfusion of a mouse with a hydrogel solution and clarification of the tissue with a combination of clearing reagents. Spinal cord tissue was cleared in just under two weeks, and the subsequent immunofluorescent staining against serotonin was completed in less than ten days. With a multi-photon fluorescent microscope, the tissue was scanned and a 3D image was reconstructed using Osirix software.

Supraspinal projections are important for pain perception and other behaviors, and serotonergic fibers are one of these fiber systems. The present study focused on the application of the combined CLARITY/CUBIC protocol to the mouse spinal cord in order to investigate the termination of these serotonergic fibers.

הדמית סיבי serotonergic בחוט שדרת עכבר שימוש CLARITY / טכניקה מעוקבת

Long descending fibers to the spinal cord are essential for locomotion, pain perception, and other behaviors. The fiber termination pattern in the spinal ...

Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System

During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or postmitotic neurons. Later, some progenitors switch to a gliogenic fate, adding to the astrocyte and oligodendrocyte populations. Using time-lapse video-microscopy of primary cerebral cortex cell cultures, it is possible to study the cellular and molecular mechanisms controlling the mode of cell division and cell cycle parameters of progenitor cells. Similarly, the fate of postmitotic cells can be examined using cell-specific fluorescent reporter proteins or post-imaging immunocytochemistry. More importantly, all these features can be analyzed at the single-cell level, allowing the identification of progenitors committed to the generation of specific cell types. Manipulation of gene expression can also be performed using viral-mediated transfection, allowing the study of cell-autonomous and non-cell-autonomous phenomena. Finally, the use of fusion fluorescent proteins allows the study of symmetric and asymmetric distribution of selected proteins during division and the correlation with daughter cells fate. Here, we describe the time-lapse video-microscopy method to image primary cerebral cortex murine cells for up to several days and analyze the mode of cell division, cell cycle length and fate of newly generated cells. We also describe a simple method to transfect progenitor cells, which can be applied to manipulate genes of interest or simply label cells with reporter proteins.

Live imaging is a powerful tool to study cellular behaviors in real time. Here, we describe a protocol for time-lapse video-microscopy of primary cerebral cortex cells that allows a detailed examination of the phases enacted during the lineage progression from primary neural stem cells to differentiated neurons and glia.

חיים הדמיה של קליפת המוח ראשי תאים באמצעות מערכת תרבות 2D

During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or ...

Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this challenging. Because the ventricles are negative spaces located deep within the brain, the only way to understand their anatomy is by appreciating their boundaries formed by related structures. Looking at a 2D representation of these spaces, in any of the cardinal planes, will not enable visualisation of all of the structures that form the boundaries of the ventricles. Thus, using 2D sections alone requires students to compute their own mental image of the 3D ventricular spaces. The aim of this study was to develop a reproducible method for dissecting the human brain to create an educational resource to enhance student understanding of the intricate relationships between the ventricles and periventricular structures. To achieve this, we created a video resource that features a step-by-step guide using a fiber dissection method to reveal the lateral and third ventricles together with the closely related limbic system and basal ganglia structures. One of the advantages of this method is that it enables delineation of the white matter tracts that are difficult to distinguish using other dissection techniques. This video is accompanied by a written protocol that provides a systematic description of the process to aid in the reproduction of the brain dissection. This package offers a valuable anatomy teaching resource for educators and students alike. By following these instructions educators can create teaching resources and students can be guided to produce their own brain dissection as a hands-on practical activity. We recommend that this video guide be incorporated into neuroanatomy teaching to enhance student understanding of the morphology and clinical relevance of the ventricles.

This paper demonstrates the effective use of a fiber dissection method to reveal the superficial white matter tracts and periventricular structures of the human brain, in three-dimensional space, to aid student comprehension of ventricular morphology.

חקר החלל העמוק - לחשוף את האנטומיה של מבנים Periventricular כדי לחשוף את החדרים הצדדיים של המוח האנושי

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this ...

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.

הקלטות סימולטני של פוטנציאל קורטיקלית שדה מקומי, רל, Electromyogram נושם קצב מעכברוש נעה בחופשיות

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

Combined Transcranial Magnetic Stimulation and Electroencephalography of the Dorsolateral Prefrontal Cortex

Transcranial magnetic stimulation (TMS) is a non-invasive method that produces neural excitation in the cortex by means of brief, time-varying magnetic field pulses. The initiation of cortical activation or its modulation depends on the background activation of the neurons of the cortical region activated, the characteristics of the coil, its position and its orientation with respect to the head. TMS combined with simultaneous electrocephalography (EEG) and neuronavigation (nTMS-EEG) allows for the assessment of cortico-cortical excitability and connectivity in almost all cortical areas in a reproducible manner. This advance makes nTMS-EEG a powerful tool that can accurately assess brain dynamics and neurophysiology in test-retest paradigms that are required for clinical trials. Limitations of this method include artifacts that cover the initial brain reactivity to stimulation. Thus, the process of removing artifacts may also extract valuable information. Moreover, the optimal parameters for dorsolateral prefrontal (DLPFC) stimulation are not fully known and current protocols utilize variations from the motor cortex (M1) stimulation paradigms. However, evolving nTMS-EEG designs hope to address these issues. The protocol presented here introduces some standard practices for assessing neurophysiological functioning from stimulation to the DLPFC that can be applied in patients with treatment resistant psychiatric disorders that receive treatment such as transcranial direct current stimulation (tDCS), repetitive transcranial magnetic stimulation (rTMS), magnetic seizure therapy (MST) or electroconvulsive therapy (ECT).

The protocol presented here is for TMS-EEG studies utilizing intracortical excitability test-retest design paradigms. The intent of the protocol is to produce reliable and reproducible cortical excitability measures for assessing neurophysiological functioning related to therapeutic interventions in the treatment of neuropsychiatric diseases such as major depression.

גירוי מגנטי טראנס המשולבת ואת אלקטרואנצפלוגרם של קליפת המוח הקדם חזיתית דורסולטרלי

Transcranial magnetic stimulation (TMS) is a non-invasive method that produces neural excitation in the cortex by means of brief, time-varying magnetic ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

השרשה כרונית של מערך שלם-קורטיקליים Electrocorticographic ב המרמוסט נפוצות

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...