אנו מודים לחברים של רובינסון ואח &#39;. 12 על תרומתם לכתב היד שממנו הפרוטוקול זה נגזר באופן חלקי.

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The authors declare that they have no competing financial interests.

פרוטוקול זה מתאר כיצד ליישם גישת pharmacogenetic (DREADD) כדי לחקור כיצד אזור ספציפי במוח תורם למשימה למידה מורכבת רב-שלב. עם היכולת באופן זמני ומרחוק כדי להשתיק את הפעילות עצבית באזורי המוח הבדידים פני שלבים של למידה, זה שילוב של גישות מספק פלטפורמה לחקור מגוון רחב של התנהגויו?...

אתגר מרגש בתחום מדעי מוח התנהגות הוא לקבוע את מצעים העצביים של התנהגויות מורכבות. מספר הטכניקות כגון נגעים קבועים, איון מוח זמני באמצעות שתלי cannulae וoptogenetics להיות מועסק לזהות את תרומתם של אזורים במוח הבדידים למשנה של התנהגויות מורכבות. בעוד גישות אלה להודיע ​​ההבנה ספציפיות האזורי שלנו בלמידה, כל טכניקה היא לא בלי מגבלות. באופן ספציפי, נגעים קבועים מתבצעים בדרך כלל לפני בדיקות התנהגותיות, כך ההשפעות שלהם נמצאות בכל משך הזמן של הפרדיגמה. מחקרי Cannulation שכרוכים בהצגת inactivator טווח קצר עצבי (למשל, tetrodotoxin) יכול לייצר נזק משמעותי לרקמת מוח ויכול לגרום ללחץ בנושאים רק לפני בדיקות התנהגותיות. יתר על כן, איון באמצעות cannulation מוגבל לאזור של רקמה המקיף אתקצה cannulae. לבסוף, בעוד optogenetics מציע מגוון רחב של גמישות לשליטה הזמנית של פעילות באזורי מוח מסוימים, זה העלות ותובענית מבחינה טכנית. ניתן להתגבר על המגבלות האלה באמצעות גישה pharmacogenetic (מעצב-הופעל באופן בלעדי על ידי מעצב-בסמים רצפטורים, DREADDs) 1,2. חשוב לציין, בזמן שמושג Pharmacogenetics הוא מתוחכם, ביצוע הטכניקה הוא פשוט. בדומה לשיטות ניתוחיות stereotaxic מסורתיים הכוללות עירוי של רעלן (למשל, NMDA, חומצת ibotenic) לאזורים במוח בדידים, טכניקה זו כרוכה בהחדרת וירוס adeno הקשורים (AAV) שמכיל קטע DNA לקולטן G- חלבון מעכב שונה בשילוב (HM4Di; קולט המעצב) לאזור של עניין של מכרסמים מעבדה סטנדרטיים (ראה איור 1). הווקטור הנגיפי מכיל גם כתב ניאון (mcitrine). ברגע שהתאגד בלתאים, קולט המעצב (וחלבון כתב) הם מקסימאלי הביעו ~ 3 שבועות לאחר עירוי-ויכולים להיות מופעל באופן סלקטיבי במשך 2-5 שעות על ידי ממשל מערכתי של תרופת מעצב אחר אינרטי מבחינה ביולוגית, קלוזאפין-N-תחמוצת (CNO) 1 , 3. בגלל ניסוי ניחן בשליטה זמנית מדויקת, עדיין רחוקה מעל הפעילות עצבית באזורים מסוימים במוח, Pharmacogenetics משלב טוב במיוחד עם פרדיגמות התנהגות שנערכות בשלבים מרובים. בדוגמא זו, התרומה של קליפת retrosplenial (RSC) ללמידה בהשוואה לתפקידה בלמידה הפבלובית-גירוי גירוי, לעומת זאת, השילוב של גישות הוא גם מתאים לכל מספר של שאלות שמבקשים לזהות כיצד ספציפי אזורים במוח תורמים ל התנהגות מורכבת. בנוסף, בזמן שאינו מתואר בפרוטוקול הנוכחי, ניתן להשתמש בגישות ויראלי ומהונדסות כדי להשיג ביטוי DREADD סוג ספציפי תא 2. כפי שים הגלום בפרדיגמות התנהגות שכרוכות תרופתיים ו / או סוגים אחרים של מניפולציות ניסוייות, שיקול זהיר של תכנון ניסוי וניתוח כמותי שלאחר מכן נדרש כאשר העסקת גישת DREADD. הנסיינים חדשים לגישת DREADD מכונים סקירה מקיפה של טכנולוגית DREADD הנוכחית 2. בכל יום, אורגניזמים ללמוד על גירויים חדשים ואירועים ומערכות היחסים שלהם אחד לשני. גם בסביבה מוכרת, כגון בית, אחד הוא מהיר כדי לזהות שינויים במערכות היחסים בין גירויים, כי שינויים אלה עשויים להיות חזוי של אירועים משמעותיים. גירוי-גירוי כזה (כלומר, יחסים) למידה כרוכה איחוד של גירויים מרובים ובאופן מסורתי נקשר עם ההיפוקמפוס, שמתגורר במרכז בתוך האונה הטמפורלית מדיאלית 4. עם זאת, ההיפוקמפוס אינו קיימת ולא לפעול בבידוד; אזורים בקליפת המוח הן בתוך ומחוץצד של האונה הטמפורלית מדיאלית לספק מידע חושי קריטי להיווצרות בהיפוקמפוס 5-7. מחקרי נגע קבועים מסורתיים לספק ראיות משכנעות למעורבותם של מספר האזורים בקליפת המוח (למשל, הקורטקס retrosplenial, postrhinal וentorhinal) בלמידה בהיפוקמפוס תלוי אך מוגבלים ביכולתם להבחין בתפקיד של אזור מסוים בשלבים בדידים למידת 8-10. הפרוטוקול הנוכחי בוחן את ההשערה שRSC הכרחי ללמידת גירוי-גירוי על ידי השתקת RSC בשלב אחד של פרדיגמה נפשית מראש חושי 3-שלב 11,12. בקצרה, חולדות לקבל עירויים של AAV שמכיל את הקולטן המעצב ו~ 3 שבועות לאחר מכן מנוהלים סמים מעצב (CNO) 30 דקות לפני תחילת בדיקות התנהגותיות. בפרוטוקול הנוכחי, חולדות ניסוי לקבל CNO במהלך השלב הראשון של בדיקות (כאשר ליר גירוי-גירוינינג מתרחש) והם מקבלים רכב במהלך 2 השלבים הבאים של בדיקות. כדי לשלוט על השפעות מכוונות של CNO על התנהגות, להחדיר חולדות עם קולט המעצב (hM4Di) ולהזריק עם רכב במקום CNO. כדי להסביר את תופעות כלליות של עירוי נגיפי וביטוי הקולטן, להחדיר וירוס שליטה שאינו מכיל את הקולטן המעצב ולנהל CNO. מספר קפסיד השונים של AAV משמש כדי לספק חומר גנטי. הנחיות NIH הנוכחיות למחקר מעורב רקומביננטי או מולקולות סינטטיים שומרת AAV ש( כל קפסיד) ומבני AAV רקומביננטי או סינטטיים, שבו transgene לא לקודד או מוצר גן פוטנציאל סרטני או מולקולת הרעלן ומיוצרים בהעדר וירוס עוזר, דורש BSL-1 אמצעי זהירות (נספח B-1. סיכון קבוצת 1 (RG1) סוכנים) 13. מספר ביקורות הנוגעות למבנה AAV, שירות ובטיחות זמין 14,15. יש לציין, אם כי, בשל חששות הנוגעים ל16,17 ופוטנציאל מנגנונים אפשריים רבייה מסרטנת 18-20 במכרסמים ל, חלק ממוסדות דורשים שימוש ב- 2 מנהלת הליגה אמצעי זהירות בעת עבודה עם AAV. ודא מנהלת הליגה המתאימה לפני השימוש על ידי התייעצות עם ועדות פיקוח במוסדות בודדים שבי המחקר ייערך, המרכז לבקרת מחלות והנחיות NIH למחקר מעורב רקומביננטי DNA מולקולות 13 בעת שימוש בוקטורים ויראליים למניפולציה גנטית בארצות הברית. הגנה אישית, הכשרת חוקר, בלימת וקטור, טיהור, סילוק חומרי חיטוי, ובעלי חיים לאחר הזרקת דרישות דיור שצוינו על ידי הנחיות אלה. בנוסף, להתייעץ ועיקבו אחרי מוסדי טיפול בבעלי חיים מתאים ובהתאם להנחיות וועדה או הנחיות ועדת ביקורת מוסדיות שווה ערך על מנת להבטיח את הטיפול, הממשל וסילוק של AAV הבטוח.

<table border="0" cellpadding="0" cellspacing="0" >
	
	
	<tbody>
		<tr >
			<td >Male, Long Evans Rats, 55-60 d</td>
			<td >Hilltop Lab Animals Inc</td>
			<td ></td>
			<td ></td>
		</tr>
		<tr >
			<td >rAAV8/hSyn-HA-hM4D(Gi)-IRES-mCitrine</td>
			<td >Virus Vector Core</td>
			<td ></td>
			<td>Caution:&#160; This is a BSL-1 compound</td>
		</tr>
		<tr >
			<td >rAAV8/hSyn-GFP</td>
			<td >Virus Vector Core</td>
			<td ></td>
			<td>Caution:&#160; This is a BSL-1 compound</td>
		</tr>
		<tr >
			<td >Clozapine-N-Oxide</td>
			<td >R&#38;D Systems</td>
			<td >4936-10</td>
			<td>Designer Drug</td>
		</tr>
		<tr >
			<td >Rat Cage lid (Polycarbonate)</td>
			<td >Alternative Design&#160;</td>
			<td >FT 8XL-PC</td>
			<td>Used to cover animal cages 48-72 hours post infusion</td>
		</tr>
		<tr >
			<td >Filer Paper (Replacement)</td>
			<td >Alternative Design&#160;</td>
			<td >FP-R-1018XAD</td>
			<td>Filter paper that goes with cage lids</td>
		</tr>
		<tr >
			<td >Table Top Vise</td>
			<td >JETS</td>
			<td >2201-265</td>
			<td>For holding microscentrifuge tubes containing AAV in the hood</td>
		</tr>
		<tr >
			<td >Medline Biohazard liners&#160;</td>
			<td >Staples</td>
			<td >113444</td>
			<td>Biohazard Trash Bags</td>
		</tr>
		<tr >
			<td >United Solutions 34 gallon rectangular wheeled trashcan with hook and lock handle&#160;</td>
			<td >Amazon.com</td>
			<td ></td>
			<td>Biohazard Trash Can</td>
		</tr>
		<tr >
			<td >Isoflurane, 100 mL</td>
			<td >Patterson Veterinary Supply Inc.&#160;</td>
			<td >07-890-8540</td>
			<td>Anesthetic</td>
		</tr>
		<tr >
			<td >Dual Small Animal Stereotaxic with Digital Display Readout Console</td>
			<td >David Kopf</td>
			<td >Model 942</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >Non-rupture Ear Bars, set of 2 (Rat)</td>
			<td >David Kopf</td>
			<td >Model 955</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >Anesthesia Mask (Rat)</td>
			<td >David Kopf</td>
			<td >Model 906</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >High speed Stereotaxic drill includes table top motor controller, foot pedal, handpiece, stereotaxic handpiece holder</td>
			<td >David Kopf</td>
			<td >Model 1474</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >Microdrill burrs, 0.9 mm</td>
			<td >Fine Science Tools Inc</td>
			<td >19007-09</td>
			<td>Surgical supply</td>
		</tr>
		<tr >
			<td >Automated Syringe pump with Micro4 Controller&#160;</td>
			<td >David Kopf</td>
			<td >Model UMP3-1</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >Pro-animal detachable Ceramic Blade Clipper Kit</td>
			<td >Ahdis</td>
			<td >21420</td>
			<td>Surgical supply</td>
		</tr>
		<tr >
			<td >Betadine Skin Cleanser</td>
			<td >Perdue&#160; Products L.P</td>
			<td >67618-149-04</td>
			<td>Surgical supply</td>
		</tr>
		<tr >
			<td >Triple Antiobiotic Ointment</td>
			<td >Medline Supply</td>
			<td >53329-087-01</td>
			<td>Surgical supply</td>
		</tr>
		<tr >
			<td >Puralube Vet Ointment</td>
			<td >Only Veterinary Supply</td>
			<td >17033-211-38</td>
			<td>Surgical supply</td>
		</tr>
		<tr >
			<td >Dino-lite&#160;</td>
			<td >Microscope</td>
			<td >AD7013MTL</td>
			<td>An alternative to the traditional disection scope</td>
		</tr>
		<tr >
			<td >Dino-lite Rigid TableTop Boom Stand</td>
			<td >Microscope</td>
			<td >MS36B</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >28 Gauge 10uL Syringe</td>
			<td >Hamilton&#160;</td>
			<td >80308-701SN</td>
			<td>Surgical equipment</td>
		</tr>
		<tr >
			<td >Extra Tall MDF Sound attenuating Cubicle</td>
			<td >Med Associates, Inc</td>
			<td >ENV-018MD</td>
			<td>22&#39;Wx22&#34;Hx16&#34;D</td>
		</tr>
		<tr >
			<td >Extra Tall Modular Test Chamber</td>
			<td >Med Associates, Inc</td>
			<td >ENV-007</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Stainless Steel Grid Floor&#160;</td>
			<td >Med Associates, Inc</td>
			<td >ENV-005</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >House Light</td>
			<td >Med Associates, Inc</td>
			<td >ENV-215M</td>
			<td>Used as the house light and stimulus light</td>
		</tr>
		<tr >
			<td >Modular Pellet Dispenser&#160;</td>
			<td >Med Associates, Inc</td>
			<td >ENV-203M-45</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Pellet Recetacle, Cup Type</td>
			<td >Med Associates, Inc</td>
			<td >ENV-200R1M</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Head Entry Detector for Rat</td>
			<td >Med Associates, Inc</td>
			<td >ENV-254-CB</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Dustless precision food pellets, 45 mg</td>
			<td >Bio-Serv</td>
			<td >F0165</td>
			<td >Behavioral supply</td>
		</tr>
		<tr >
			<td >Cage Speaker for Rat Chamber</td>
			<td >Med Associates, Inc</td>
			<td >ENV-224AM</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Programmable Audio Generator&#160;</td>
			<td >Med Associates, Inc</td>
			<td >ANL-926</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Smart Ctrl 8 Input/16 output Package</td>
			<td >Med Associates, Inc</td>
			<td >DIG-716P2</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Large Table Top Cabinet and Power Supply</td>
			<td >Med Associates, Inc</td>
			<td >SG-6510D</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >PCI Interface Package</td>
			<td >Med Associates, Inc</td>
			<td >DIG-700P2-R2</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >MED Intel core Computer Pkg with X Pro 19&#34; Monitor</td>
			<td >Med Associates, Inc</td>
			<td >COM-103V</td>
			<td>Behavioral equipment</td>
		</tr>
		<tr >
			<td >Paraformaldehyde (grannular), 1 kg</td>
			<td >Electron Microsopy Sciences</td>
			<td >19210</td>
			<td>Hazard:&#160; carcinogen, weigh in hood</td>
		</tr>
		<tr >
			<td >Rabbit Monoclonal antibody (HA-Tag)</td>
			<td >Cell Signaling Technologies&#160;</td>
			<td >3724S</td>
			<td>Histology reagent</td>
		</tr>
		<tr >
			<td >XP Rabbit monoclonal antibody (GFP)</td>
			<td >Cell Signaling Technologies&#160;</td>
			<td >2956S</td>
			<td>Histology reagent</td>
		</tr>
		<tr >
			<td >Anti-Rabbit IgG</td>
			<td >Cell Signaling Technologies&#160;</td>
			<td >4412S</td>
			<td>Histology supplies</td>
		</tr>
		<tr >
			<td >Superfrost Plus slides&#160;</td>
			<td >VWR international</td>
			<td >483111-703</td>
			<td>Histology supplies</td>
		</tr>
	</tbody>
</table>

a method for remotely silencing neural activity in rodents during discrete phases of learning

פרוטוקול זה מתאר כיצד באופן זמני ומרחוק כדי להשתיק את הפעילות עצבית באזורי המוח בדידים בעוד בעלי החיים עוסקים במשימות למידה וזיכרון. הגישה משלבת Pharmacogenetics (מעצב-רצפטורים-הופעל באופן בלעדי על ידי מעצב-בסמים) עם הפרדיגמה התנהגות (נפשית מראש חושי) שנועד להבחין בין צורות שונות של למידה. באופן ספציפי, משלוח בתיווך נגיפי משמש להביע קולט G-חלבון מעכב מהונדס גנטי בשילוב (מעצב קולט) לאזור במוח בדיד במכרסמים. שלושה שבועות לאחר מכן, כאשר רמות ביטוי קולט מעצב הן גבוהות, סוכן תרופתי (מעצב התרופות) מנוהל מערכתי 30 דקות לפני פגישת התנהגות ספציפית. יש תרופת זיקה לקולטן מעצב ובכך גורמת לעיכוב של נוירונים המבטאים את הקולטן המעצב, אבל אחרת אינרטי מבחינה ביולוגית. האזור במוח נשאר מושתק במשך 2-5 שעות (depenדינג על המינון ודרך המתן). עם השלמת הפרדיגמה ההתנהגות, רקמת המוח נבחנת למיקום נכון וביטוי הקולטן. גישה זו שימושית במיוחד לקביעת התרומה של אזורים במוח אדם לרכיבים ספציפיים של התנהגות וניתן להשתמש בו על פני כל מספר של פרדיגמות התנהגות.

תוצאות התנהגות עם השלמת הניסוי, את האפקטיביות של איון הזמני ספציפי לאזור יש להעריך כמותית ואיכותית. הדוגמא הנוכחית כרוכה הפרדיגמה 3-שלב התנהגות (נפשית מראש חושי), שבו היה מנוהל CNO כדי להחליש פעילות עצבית בRSC במהלך פגישות אכשור קד לבח...

Watch this Scientific Journal Video about A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning at JoVE.com

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

שיטה להשתקה מרחוק עצבית פעילות במכרסמים בשלבים בדידים של למידה

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

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Research

JoVE Journal

Behavior

14.6K Views.  Oberlin College. This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

Video: A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

Meal Duration as a Measure of Orofacial Nociceptive Responses in Rodents

A lengthening in meal duration can be used to measure an increase in orofacial mechanical hyperalgesia having similarities to the guarding behavior of humans with orofacial pain. To measure meal duration unrestrained rats are continuously kept in sound attenuated, computerized feeding modules for days to weeks to record feeding behavior. These sound-attenuated chambers are equipped with chow pellet dispensers. The dispenser has a pellet trough with a photobeam placed at the bottom of the trough and when a rodent removes a pellet from the feeder trough this beam is no longer blocked, signaling the computer to drop another pellet. The computer records the date and time when the pellets were taken from the trough and from this data the experimenter can calculate the meal parameters. When calculating meal parameters a meal was defined based on previous work and was set at 10 min (in other words when the animal does not eat for 10 min that would be the end of the animal&#39;s meal) also the minimum meal size was set at 3 pellets. The meal duration, meal number, food intake, meal size and inter-meal interval can then be calculated by the software for any time period that the operator desires. Of the feeding parameters that can be calculated meal duration has been shown to be a continuous noninvasive biological marker of orofacial nociception in male rats and mice and female rats. Meal duration measurements are quantitative, require no training or animal manipulation, require cortical participation, and do not compete with other experimentally induced behaviors. These factors distinguish this assay from other operant or reflex methods for recording orofacial nociception.

A lengthening in meal duration represents orofacial nociceptive behavior in rodents similar to the guarding behavior of humans with orofacial pain. Eating is a behavior that requires no training or animal manipulation, requires cortical participation, and is not competing with other experimentally induced behaviors, distinguishing this assay from alternative reflex or operant measurements.

משך הארוחה כמדד לתגובות הנוצ'ית האורופציאליות במכרסמים

A lengthening in meal duration can be used to measure an increase in orofacial mechanical hyperalgesia having similarities to the guarding behavior of ...

Automated Visual Cognitive Tasks for Recording Neural Activity Using a Floor Projection Maze

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have developed visual cognitive tasks for rats using the Floor Projection Maze1,2&#160;that are optimized for visual abilities of rats permitting stronger comparisons of experimental findings with other species.
In order to investigate neural correlates of learning and memory, we have integrated electrophysiological recordings into fully automated cognitive tasks on the Floor Projection Maze1,2. Behavioral software interfaced with an animal tracking system allows monitoring of the animal's behavior with precise control of image presentation and reward contingencies for better trained animals. Integration with an in vivo electrophysiological recording system enables examination of behavioral correlates of neural activity at selected epochs of a given cognitive task.
We describe protocols for a model system that combines automated visual presentation of information to rodents and intracranial reward with electrophysiological approaches. Our model system offers a sophisticated set of tools as a framework for other cognitive tasks to better isolate and identify specific mechanisms contributing to particular cognitive processes.

We describe protocols for training rats for chronic electrophysiological recordings in fully automated cognitive tasks on a Floor Projection Maze.

משימות אוטומטיות חזותית קוגניטיבית להקלטה עצבית פעילות באמצעות הקרנה מקיר לקיר מבוך

Neuropsychological tasks used in primates to investigate mechanisms of learning and memory are typically visually guided cognitive tasks. We have ...

Measuring Neural and Behavioral Activity During Ongoing Computerized Social Interactions: An Examination of Event-Related Brain Potentials

Social exclusion is a complex social phenomenon with powerful negative consequences. Given the impact of social exclusion on mental and emotional health, an understanding of how perceptions of social exclusion develop over the course of a social interaction is important for advancing treatments aimed at lessening the harmful costs of being excluded. To date, most scientific examinations of social exclusion have looked at exclusion after a social interaction has been completed. While this has been very helpful in developing an understanding of what happens to a person following exclusion, it has not helped to clarify the moment-to-moment dynamics of the process of social exclusion. Accordingly, the current protocol was developed to obtain an improved understanding of social exclusion by examining the patterns of event-related brain activation that are present during social interactions. This protocol allows greater precision and sensitivity in detailing the social processes that lead people to feel as though they have been excluded from a social interaction. Importantly, the current protocol can be adapted to include research projects that vary the nature of exclusionary social interactions by altering how frequently participants are included, how long the periods of exclusion will last in each interaction, and when exclusion will take place during the social interactions. Further, the current protocol can be used to examine variables and constructs beyond those related to social exclusion. This capability to address a variety of applications across psychology by obtaining both neural and behavioral data during ongoing social interactions suggests the present protocol could be at the core of a developing area of scientific inquiry related to social interactions.

Research on social exclusion has grown tremendously in recent years. As the field expands, it is imperative to develop sophisticated methodologies allowing for the simultaneous measurement of neural and behavioral outcomes during social exclusion. This protocol utilizes event-related brain potentials to record ongoing neural activity during computerized social interactions.

מדידה עצבית והתנהגות פעילות במהלך אינטראקציות חברתיות ממוחשבות מתמשכות: בחינה של פוטנציאל מוח הקשור לאירוע

Social exclusion is a complex social phenomenon with powerful negative consequences. Given the impact of social exclusion on mental and emotional health, ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

זוגי H-המבוך: מבחן חזק התנהגות למידה וזיכרון במכרסמים

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

Quantifying Learning in Young Infants: Tracking Leg Actions During a Discovery-learning Task

Task-specific actions emerge from spontaneous movement during infancy. It has been proposed that task-specific actions emerge through a discovery-learning process. Here a method is described in which 3-4 month old infants learn a task by discovery and their leg movements are captured to quantify the learning process. This discovery-learning task uses an infant activated mobile that rotates and plays music based on specified leg action of infants. Supine infants activate the mobile by moving their feet vertically across a virtual threshold. This paradigm is unique in that as infants independently discover that their leg actions activate the mobile, the infants&#8217; leg movements are tracked using a motion capture system allowing for the quantification of the learning process. Specifically, learning is quantified in terms of the duration of mobile activation, the position variance of the end effectors (feet) that activate the mobile, changes in hip-knee coordination patterns, and changes in hip and knee muscle torque. This information describes infant exploration and exploitation at the interplay of person and environmental constraints that support task-specific action. Subsequent research using this method can investigate how specific impairments of different populations of infants at risk for movement disorders influence the discovery-learning process for task-specific action.

A method is described in which 3-4 month old infants learn a task by discovery and their leg movements are captured to quantify the learning process.

כימות למידה בתינוקות צעירים: פעולות מעקב רגלו במהלך משימה למידת גילוי

Task-specific actions emerge from spontaneous movement during infancy. It has been proposed that task-specific actions emerge through a discovery-learning ...

Asymmetric Walkway: A Novel Behavioral Assay for Studying Asymmetric Locomotion

Behavioral assays are commonly used for the assessment of sensorimotor impairment in the central nervous system (CNS). The most sophisticated methods for quantifying locomotor deficits in rodents is to measure minute disturbances of unconstrained gait overground (e.g., manual BBB score or automated CatWalk). However, cortical inputs are not required for the generation of basic locomotion produced by the spinal central pattern generator (CPG). Thus, unconstrained walking tasks test locomotor deficits due to motor cortical impairment only indirectly. In this study, we propose a novel, precise foot-placement locomotor task that evaluates cortical inputs to the spinal CPG. An instrumented peg-way was used to impose symmetrical and asymmetrical locomotor tasks mimicking lateralized movement deficits. We demonstrate that shifts from equidistant inter-stride lengths of 20% produce changes in the forelimb stance phase characteristics during locomotion with preferred stride length. Furthermore, we propose that the asymmetric walkway allows for measurements of behavioral outcomes produced by cortical control signals. These measures are relevant for the assessment of impairment after cortical damage.

Here, we present a protocol to quantify precise stepping in rodents. Cortical and the spinal central pattern generator signals are required for precise foot-placement during obstructed locomotion. We report here the novel constrained walking task that directly examines precise stepping behavior.

א-סימטרי טיילת: רומן התנהגות Assay עבור לימוד אסימטרית תנועה

Behavioral assays are commonly used for the assessment of sensorimotor impairment in the central nervous system (CNS). The most sophisticated methods for ...

Using the Activity-based Anorexia Rodent Model to Study the Neurobiological Basis of Anorexia Nervosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A challenging illness to treat, due to the lack of understanding of the underlying neurobiology, AN has the highest mortality rate among psychiatric illnesses. To address this need, neuroscientists are using an animal model to study how neural circuits may contribute toward vulnerability to AN and may be affected by AN. Activity-based anorexia (ABA) is a bio-behavioral phenomenon described in rodents that models the key symptoms of anorexia nervosa. When rodents with free access to voluntary exercise on a running wheel experience food restriction, they become hyperactive &#8211; running more than animals with free access to food. Here, we describe the procedures by which ABA is induced in adolescent female C57BL/6 mice. On postnatal day 36 (P36), the animal is housed with access to voluntary exercise on a running wheel. After 4 days of acclimation to the running wheel, on P40, all food is removed from the cage. For the next 3 days, food is returned to the cage (allowing animals free food access) for 2 hr daily. After the fourth day of food restriction, free access to food is returned and the running wheel is removed from the cage to allow the animals to recover. Continuous multi-day analysis of running wheel activity shows that mice become hyperactive within 24 hr following the onset of food restriction. The mice run even during the limited time during which they have access to food. Additionally, the circadian pattern of wheel running becomes disrupted by the experience of food restriction. We have been able to correlate neurobiological changes with various aspects of the animals&#8217; wheel running behavior to implicate particular brain regions and neurochemical changes with resilience and vulnerability to food-restriction induced hyperactivity.

Here we present a protocol to induce activity-based anorexia (ABA) in female adolescent mice. ABA is a condition of hyperactivity evoked by imposing food restriction on rodents with access to a running wheel. This phenomenon is being used as a model to study the underlying neurobiology of anorexia nervosa.

באמצעות מודל אנורקסיה מכרסמים מבוסס פעילות לחקר הבסיס הנוירוביולוגי של אנורקסיה נרבוזה

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A ...

A Novel Experimental and Analytical Approach to the Multimodal Neural Decoding of Intent During Social Interaction in Freely-behaving Human Infants

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental paradigms and analysis tools have been limited to constrained laboratory tasks and contexts due to technical limitations imposed by the available set of measuring and analysis techniques and the age of the subjects. These limitations severely limit the study of developmental neural dynamics and associated neural networks engaged in cognition, perception and action in infants performing &#8220;in action and in context&#8221;. This protocol presents a novel approach to study infants and young children as they freely organize their own behavior, and its consequences in a complex, partly unpredictable and highly dynamic environment. The proposed methodology integrates synchronized high-density active scalp electroencephalography (EEG), inertial measurement units (IMUs), video recording and behavioral analysis to capture brain activity and movement non-invasively in freely-behaving infants. This setup allows for the study of neural network dynamics in the developing brain, in action and context, as these networks are recruited during goal-oriented, exploration and social interaction tasks.

This protocol presents a novel methodology for the neural decoding of intent from freely-behaving infants during unscripted social interaction with an actor. Neural activity is acquired using non-invasive high-density active scalp electroencephalography (EEG). Kinematic data is collected with inertial measurement units and supplemented with synchronized video recording.

גישה חדשנית ניסויית ואנליטית למולטימודליות עצביות פענוח כוונות במהלך אינטראקציה חברתית באופן חופשי על-מתנהג תינוקות אנושיים

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental ...

Feeding Experimentation Device (FED): Construction and Validation of an Open-source Device for Measuring Food Intake in Rodents

Food intake measurements are essential for many research studies. Here, we provide a detailed description of a novel solution for measuring food intake in mice: the Feeding Experimentation Device (FED). FED is an open-source system that was designed to facilitate flexibility in food intake studies. Due to its compact and battery powered design, FED can be placed within standard home cages or other experimental equipment. Food intake measurements can also be synchronized with other equipment in real-time via FED's transistor-transistor logic (TTL) digital output, or in post-acquisition processing as FED timestamps every event with a real-time clock. When in use, a food pellet sits within FED's food well where it is monitored via an infrared beam. When the pellet is removed by the mouse, FED logs the timestamp onto its internal secure digital (SD) card and dispenses another pellet. FED can run for up to 5 days before it is necessary to charge the battery and refill the pellet hopper, minimizing human interference in data collection. Assembly of FED requires minimal engineering background, and off-the-shelf materials and electronics were prioritized in its construction. We also provide scripts for analysis of food intake and meal patterns. Finally, FED is open-source and all design and construction files are online, to facilitate modifications and improvements by other researchers.

Feeding Experimentation Device FED: Construction and Validation of an Open-source Device for Measuring Food Intake in Rodents

Feeding Experimentation Device (FED) is an open-source device for measuring food intake in mice. FED can also synchronize food intake measurements with other techniques via a real-time digital output. Here, we provide a step-by-step tutorial for the construction, validation, and usage of FED.

האכלת התקני ניסויים (FED): בנייה ותיקוף של התקן קוד פתוח עבור מדידת צריכת מזון במכרסמים

Food intake measurements are essential for many research studies. Here, we provide a detailed description of a novel solution for measuring food intake in ...

A Computational Method to Quantify Fly Circadian Activity

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. Fundamental mechanisms that underlie this temporal control are widely studied using the fruit fly Drosophila melanogaster as a model organism. In flies, the clock is typically studied by analyzing multiday locomotor recording. Such a recording shows a complex bimodal pattern with two peaks of activity: a morning peak that happens around dawn, and an evening peak that happens around dusk. These two peaks together form a waveform that is very different from sinusoidal oscillations observed in clock genes, suggesting that mechanisms in addition to the clock have profound effects in producing the observed patterns in behavioral data. Here we provide instructions on using a recently developed computational method that mathematically describes temporal patterns in fly activity. The method fits activity data with a model waveform that consists of four exponential terms and nine independent parameters that fully describe the shape and size of the morning and evening peaks of activity. The extracted parameters can help elucidate the kinetic mechanisms of substrates that underlie the commonly observed bimodal activity patterns in fly locomotor rhythms.

A method to quantify the main temporal features seen in fly circadian locomotor rhythms is presented. The quantification is achieved by fitting fly activity with a multi-parametric model waveform. The model parameters describe the shape and size of the morning and evening peaks of daily activity.