Wellcome Trust עבור מימון גישה פתוחה אוניברסיטת אוקספורד. רוברט דיקון הוא חבר בקבוצה OXION אוקספורד, הממומן על ידי Wellcome Trust מענק WT084655MA.

<ol>
	<li>Cunningham, C., Deacon, R. M. J., Chan, K., Boche, D., Rawlins, J. N. P., Perry, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropathologically+distinct+prion+strains+give+rise+to+similar+temporal+profiles+ofbehavioural+deficits.">Neuropathologically distinct prion strains give rise to similar temporal profiles ofbehavioural deficits.</a> Neurobiol. Dis. 18, 258-269 (2005).</li><li>Cunningham, C., Deacon, R., Wells, H., Boche, D., Waters, S., Picanco Diniz, C., Scott, H., Rawlins, J. N. P., Perry, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+changes+characterize+early+behavioural+signs+in+the+ME7+model+of+murine+prion+disease.">Synaptic changes characterize early behavioural signs in the ME7 model of murine prion disease.</a> Eur. J. Neurosci. 17, 2147-2215 (2003).</li><li>Damiani, R., Modesto, S., Yates, A., Neveling, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Earliest+evidence+of+cynodont+burrowing.">Earliest evidence of cynodont burrowing.</a> Proc. R. Soc. Lond. B. 270, 1747-1751 (2003).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrowing+in+rodents:+a+sensitive+method+for+detecting+behavioral+dysfunction.">Burrowing in rodents: a sensitive method for detecting behavioral dysfunction.</a> Nat. Protocols. 1, 118-121 (2006).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+nest+building+in+mice.">Assessing nest building in mice.</a> Nat. Protocols. 1, 1117-1119 (2006).</li><li>Deacon, R. M. J., Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digging+in+Mice:+Marble+Burying,+Burrowing,+and+Direct+Observation+Reveal+Changes+in+Mouse+Behavior.">Digging in Mice: Marble Burying, Burrowing, and Direct Observation Reveal Changes in Mouse Behavior.</a> Mood and anxiety related phenotypes in mice. , (2009).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+hoarding+in+mice.">Assessing hoarding in mice.</a> Nat. Protocols. 1, 2828-2830 (2006).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrowing:+A+sensitive+behavioural+assay,+tested+in+five+species+of+laboratory+rodents.">Burrowing: A sensitive behavioural assay, tested in five species of laboratory rodents.</a> Behav. Brain. Res. 200, 128-133 (2009).</li><li>Deacon, R. M. J., Bannerman, D. M., Kirby, B. P., Croucher, A., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+cytotoxic+hippocampal+lesions+in+mice+on+a+cognitive+test+battery.">Effects of cytotoxic hippocampal lesions in mice on a cognitive test battery.</a> Behav. Brain. Res. 133, 57-68 (2002).</li><li>Deacon, R. M. J., Brook, R. C., Meyer, D., Haeckel, O., Ashcroft, F. M., Miki, T., Seino, S., Liss, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+of+mice+lacking+the+KATP+channel+subunit+Kir6.2.">Behavioral phenotyping of mice lacking the KATP channel subunit Kir6.2.</a> Physiol. Behav. 87, 723-733 (2006).</li><li>Deacon, R. M. J., Croucher, A., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+cytotoxic+lesion+effects+on+species-typical+behaviors+in+mice.">Hippocampal cytotoxic lesion effects on species-typical behaviors in mice.</a> Behav. Brain. Res. 132, 203-213 (2002).</li><li>Deacon, R. M. J., Penny, C., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+medial+prefrontal+cortex+cytotoxic+lesions+in+mice.">Effects of medial prefrontal cortex cytotoxic lesions in mice.</a> Behav. Brain. Res. 139, 139-155 (2003).</li><li>Deacon, R. M. J., Raley, J. M., Perry, V. H., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrowing+into+prion+disease.">Burrowing into prion disease.</a> Neuroreport. 12, 2053-2057 (2001).</li><li>Deacon, R. M. J., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+lesions,+species-typical+behaviours+and+anxiety+in+mice.">Hippocampal lesions, species-typical behaviours and anxiety in mice.</a> Behav. Brain. Res. 156, 241-249 (2005).</li><li>Deacon, R. M. J., Thomas, C. L., Rawlins, J. N. P., Morley, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+the+behavior+of+C57BL/6+and+C57BL/10.">A comparison of the behavior of C57BL/6 and C57BL/10.</a> Behav. Brain. Res. 179, 239-247 (2007).</li><li>Guenther, K., Deacon, R. M. J., Perry, V. H., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+behavioural+changes+in+scrapie-affected+mice+and+the+influence+of+dapsone.">Early behavioural changes in scrapie-affected mice and the influence of dapsone.</a> Eur. J. Neurosci. 14, 401-409 (2001).</li><li>Mallucci, G. R., White, F. a. r. m. e. r., Dickinson, M., Khatun, A., Powell, H., Brandner, A. D., Jefferys, S., R, J. G., Collinge, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+Cellular+Prion+Protein+Reverses+Early+Cognitive+Deficits+and+Neurophysiological+Dysfunction+in+Prion-Infected+Mice.">Targeting Cellular Prion Protein Reverses Early Cognitive Deficits and Neurophysiological Dysfunction in Prion-Infected Mice.</a> Neuron. 53, 325-335 (2007).</li><li>Russell, W. M. S., Burch, R. L. <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 principles of humane experimental technique. , (1959).</li><li>Schmid-Holmes, S., Drickamer, L. C., Robinson, A. S., Gillie, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrows+and+burrow+cleaning+behaviour+of+house+mice.">Burrows and burrow cleaning behaviour of house mice.</a> Am. Mid. Nat. 146, 53-62 (2001).</li><li>Teeling, J. L., Felton, L. M., Deacon, R. M. J., Cunningham, C., Rawlins, J. N. P., Perry, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sub-pyrogenic+systemic+inflammation+impacts+on+brain+and+behavior,+independent+of+cytokines.">Sub-pyrogenic systemic inflammation impacts on brain and behavior, independent of cytokines.</a> Brain Behav. Immun. 21, 836-850 (2007).</li></ol>

אין ניגודי אינטרסים הכריז.

<html> בדיקות אלו הינן פשוטות ביותר להפעלה אך בגדלים השפעה הן גדולות מאוד, מה שהופך אותם רגישים במיוחד לטיפולים, הבדלים זן, וכו &#39;הם תואמים את ההיבט &quot;חידוד&quot; של ראסל Burch של 3 Rs &quot;18. ואכן, ביצועים טובים במבחנים כמו נבירה כנראה מעיד על בריאות כללית טובה אצל בעלי חיים. לכן זה יכול להיות מבחן טוב עבור הערכה של רווחת בעלי חיים. החשיבות של העשרה סביבתית מכרסמים במעבדה מוכרת היטב בימים אלה. מתן חומר קינון כיום מקובל בבתים בעלי חיים רבים, ואת זה קל לעשות. ההתלהבות שבה מאורה עכברים עושה נבירה העשרה מצוין בפוטנציה, אך למרבה הצער &quot;עצמית מחדש&quot; המאורה שבה תהיה אפשרות מעשית לספק זה טרם פותחו. אם העכברים למצוא אגירה להעשיר טרם נקבע. _content &quot;&gt; חסרון אחד הוא כי הנתונים שנוצר על ידי בדיקות אלה היא בדרך כלל אי ​​- פרמטרית (לא גאוס בחלוקת), המתאימה כל כך אי - פרמטרית סטטיסטיקה יש צורך לנתח אותם, למשל את מבחן Mann-Whitney U להשוואות pairwise, Kruskal- ואליס ANOVA עבור הקבוצה מרובת נתונים. אם חזר עיצובים אמצעים המשמשים, שכן אין מבחן ANOVA מתאים שאינם פרמטרית הנתונים, התוצאות ניתן להפוך, למשל על ידי שורש ריבועי או נהלים להתחבר כדי לגרום להם לציית הפצה נורמלית יותר . המדידות של המנגנון יכול להיות שונה במקצת לאלה שצוינו בטקסט (שימושי עבור הנסיינים ארה&quot;ב) אבל ההבדלים רחב יש להימנע. עם זאת, במקרים מסוימים נסיינים אחרים עשויים למצוא מבנים שונים לעבוד טוב יותר מאלו שתוארו כאן. באשר כל המדידות התנהגותיות, העכברים צריך להיבדק כדי לראות אם רמת הפעילות שלהם הן נורמליות, כמו חוסר פעילות יהיה ברור יש detrimאפקט ental על מינים טיפוסי התנהגויות. כאשר scrapie הנגועים בעכברים הראו נבירה לקויי קינון, רמות הפעילות שלהם היו תקינות (נבדק המנגנון שדה פתוח). אכן, תקופה של היפראקטיביות מתרחשת סביב 2-4 שבועות לאחר מינים אלה טיפוסי נפגעות התנהגויות הראשון 2. ליקוי חזק המיוצר על ידי נגעים בהיפוקמפוס על מתחפרת, קינון אגירה, כל אלה הם מינים טיפוסי התנהגויות, משלים את ליקויי למידה מרחבית memory9 שהם כאלה ומבוססת ההשפעות של נגעים אלה. הם נראים שווה את ליקויי בפעילויות היומיומיות האופיינית כל כך 14 לספירה. אלו זנים טיפוסי בדיקות עשוי להתברר שימוש רב ההקרנה פרה טיפולים למחלת אלצהיימר ומחלות ניווניות אחרות 14. ברבות הפרעות המוח האנושי יכולת אובדן או מופחת לבצע ערב רגילמשימות ryday חשובה כמו תכונות אחרות של המחלה כמו ליקוי קוגניטיבי. במיוחד, הפסד זה עשוי להצריך מעורבותו של המטפלים ואת נטל כלכלי גדול כל כך קרובות זו מטילה.

assessing burrowing, nest construction, and hoarding in mice

הידרדרות ביכולת לבצע &quot;פעולות יומיומיות&quot; (ADL) הוא סימן מוקדם של מחלת אלצהיימר (AD). ההקרנה של טיפולים התנהגותיים Preclinical אפשרי לספירה כיום מתמקד בעיקר בדיקות קוגניטיביות, אשר לעתים קרובות דורש ציוד יקר והרבה זמן הניסוי. עם זאת, זיכרון אפיזודי האדם (ההיבט מושפעת באופן חמור ביותר של הזיכרון לספירה) שונה לזיכרון מכרסם, אשר נראה כי במידה רבה לא אפיזודי. לכן הדרכים הנוכחי של סינון עבור טיפולים לספירה חדשה לספירה במכרסמים סביר מהותית להצליח. גישה חדשה ההקרנה פרה יהיה לאפיין את הליגה נגד השמצה של עכברים. למזלו, כגון מספר מבחני לאחרונה פותחה באוניברסיטת אוקספורד, והנה שלוש הרגישים ביותר היטב מאופיין מוצגים. נבירה פותחה לראשונה באוקספורד 13. היא התפתחה מתוך צורך לפתח פרדיגמה אגירה העכבר. רוב לפרסםאגירה ed פרדיגמות מכרסם נדרש מקור מזון מרוחק להיות מקושרים הכלוב הביתה בקטע המחבר. זה יהיה כרוך שינוי בכלוב הביתה, כמו גם ביצוע עכבר הוכחה מעבר חיבור מקור מזון. אז זה היה נחשב אם אפשר יהיה לשים את מקור המזון בתוך הכלוב. נמצא כי אם המכולה הוצבה על הרצפה זה היה מרוקן עד הבוקר הבא., כדורי המזון, לעומת זאת, פשוט שהופקדו בערימה בפתח המכולה, ולא להציב במקום בדידה מן המיכל, כפי שניתן היה לצפות אם העכברים היו באמת לאגור אותם. מבט מקרוב הראה כי העכברים היו ביצוע חפירה (&quot;נבירה&quot;), תנועות, לא נושאת את כדורי בפיהם למקום שנבחר כפי שהם היו אם באמת לאגור אותם. 6 כופתיות מזון אינם מצע חיוני מתחפרת, עכברים שפופרות הריק יהיה מלא חול וחצץ, אפילו מצעים מלוכלכים מהכלוב שלהם. יותרושוב, הם יהיו ריקים צינור מלא גם אם ריקה מונחת לידו 8. קיימים מספר פרוטוקולים קינון בספרות. אוקספורד להציג את אחד מפשטת את ההליך יש מערכת מוגדרת היטב ניקוד על איכות הקן 5. הפרדיגמה אגירה פותחה מאוחר יותר שבו עכברים, ולא אגירה בחזרה לכלוב בית אמיתי, הותאמו החיים &quot;בסיס הבית&quot; של מנגנון אגירה. בסיס זה הבית היה מחובר צינור עשוי רשת תיל, סוף דיסטלי של שהכיל את מקור המזון. הסדר זה הוכיח התנהגות אגירה תשואה טובה, כל עוד עכברים הותאמו החיים &quot;בסיס הבית&quot; במהלך היום מותר רק כדי להזין את הצינור אגירה בלילה.

Watch this Scientific Journal Video about Assessing Burrowing, Nest Construction, and Hoarding in Mice at JoVE.com

Assessing Burrowing, Nest Construction, and Hoarding in Mice

מתחפרת, קינון אגירה הם מינים טיפוסי פעילויות עכברים בקלות לבצע במעבדה. מאמר זה מתאר כיצד הם יכולים להעריך בקלות ובזול. פרוטוקולים אלה רגישים במיוחד זן העכבר, נגעים במוח ומחלות. יתר על כן הם מהווים &quot;העשרה סביבתית&quot; לעכברים, ואת לגלם את &quot;חידוד&quot; היבט של &quot;3 Rs&quot; 18.

הערכת מתחפרת, בניית קן, ו אגירה של עכברים

Deterioration in the ability to perform "Activities of daily living" (ADL) is an early sign of Alzheimer's disease (AD). Preclinical behavioural screening ...

assessing-burrowing-nest-construction-and-hoarding-in-mice

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

31.9K Views.  University of Oxford. מתחפרת, קינון אגירה הם מינים טיפוסי פעילויות עכברים בקלות לבצע במעבדה. מאמר זה מתאר כיצד הם יכולים להעריך בקלות ובזול. פרוטוקולים אלה רגישים במיוחד זן העכבר, נגעים במוח ומחלות. יתר על כן הם מהווים "העשרה סביבתית" לעכברים, ואת לגלם את "חידוד" היבט של "3 R...

Video: Assessing Burrowing, Nest Construction, and Hoarding in Mice

Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial products are often expensive, not easily customized, and not submersible. Here we describe a method to design and build a customized, integrated circuit headstage for simultaneous 4-channel neural recording and 2-channel simulation in awake, behaving animals. The headstage is designed using a free, commercially available CAD-type design package, and can be modified easily to accommodate different scales (e.g. to add channels). A customized printed circuit board is built using surface mount resistors, capacitors and operational amplifiers to construct the unity gain source follower circuit. The headstage is made water-proof with a combination of epoxy, parafilm and a synthetic rubber putty. We have successfully used this device to record local field potentials and stimulate different brain regions simultaneously via independent channels in rats swimming in a water maze. The total cost is &lt; $30/unit and can be manufactured readily.

We present a low-cost method to design and construct a light headstage pre-amplifier system with simultaneous neural recording and stimulation capability. This device can be waterproofed for use in swimming animals.

עיצוב ובנייה של Headstage העלות האפקטיבית של גירוי עצבי סימולטני הקלטה של ​​מבוך המים

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

T-מבוך לסירוגין בכפייה של שמאל וימין משימות אפליה להערכת עבודה וזיכרון העזר עכברים

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.
The Drosophila larva is an established model system to study thermal nociception, a sensory response to potentially harmful temperatures that is evolutionarily conserved across species1,2. The advantages of Drosophila for such studies are the relative simplicity of its nervous system and the sophistication of the genetic techniques that can be used to dissect the molecular basis of the underlying biology4-6 In Drosophila, as in all metazoans, the response to noxious thermal stimuli generally involves a "nocifensive" aversive withdrawal to the presented stimulus7. Such stimuli are detected through free nerve endings or nociceptors and the amplitude of the organismal response depends on the number of nociceptors receiving the noxious stimulus8. In Drosophila, it is the class IV dendritic arborization sensory neurons that detect noxious thermal and mechanical stimuli9 in addition to their recently discovered role as photoreceptors10. These neurons, which have been very well studied at the developmental level, arborize over the barrier epidermal sheet and make contacts with nearly all epidermal cells11,12. The single axon of each class IV neuron projects into the ventral nerve cord of the central nervous system11 where they may connect to second-order neurons that project to the brain.
Under baseline conditions, nociceptive sensory neurons will not fire until a relatively high threshold is reached. The assays described here allow the investigator to quantify baseline behavioral responses or, presumably, the sensitization that ensues following tissue damage. Each assay provokes distinct but related locomotory behavioral responses to noxious thermal stimuli and permits the researcher to visualize and quantify various aspects of thermal nociception in Drosophila larvae. The assays can be applied to larvae of desired genotypes or to larvae raised under different environmental conditions that might impact nociception. Since thermal nociception is conserved across species, the findings gleaned from genetic dissection in Drosophila will likely inform our understanding of thermal nociception in other species, including vertebrates.

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.

שיטות מקומיים וכלל עולמיים של הערכת Nociception תרמי ב<em&gt; תסיסנית</em&gt; רימה

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

שיטה פשוטה וזולה לקביעה קרה רגישות והסתגלות בעכברים

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

Primary neurogenesis is a dynamic and complex process during embryonic development that sets up the initial layout of the central nervous system. During this process, a portion of neural stem cells undergo differentiation and give rise to the first populations of differentiated primary neurons within the nascent central nervous system. Several vertebrate model organisms have been used to explore the mechanisms of neural cell fate specification, patterning, and differentiation. Among these is the African clawed frog, Xenopus, which provides a powerful system for investigating the molecular and cellular mechanisms responsible for primary neurogenesis due to its rapid and accessible development and ease of embryological and molecular manipulations. Here, we present a convenient and rapid method to observe the different populations of neuronal cells within Xenopus central nervous system. Using antibody staining and immunofluorescence on sections of Xenopus embryos, we are able to observe the locations of neural stem cells and differentiated primary neurons during primary neurogenesis.

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

This article presents a convenient and rapid method for visualizing different neuronal cell populations in the central nervous system of Xenopus embryos using immunofluorescent staining on sections.

הערכת Neurogenesis יסודי<em&gt; Xenopus</em&gt; עוברים באמצעות Immunostaining

Primary neurogenesis is a dynamic and complex process during embryonic development that sets up the initial layout of the central nervous system. During ...

Assessing Spatial Learning and Memory in Small Squamate Reptiles

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, interest in the cognitive processes of nonmodel species, typically within an ecological context, has also become an emerging field of study. In particular, interest in the cognitive processes in reptiles is growing although experimental studies on reptilian cognition are sparse. The few reptilian studies that have experimentally tested for spatial learning and memory have used rodent paradigms modified for use in reptiles. However, ecologically important aspects of the physiology and behavior of this taxonomic group must be taken into account when testing for spatially based cognition. Here, we describe modifications of the dry land Barnes maze and associated testing protocol that can improve performance when probing for spatial learning and memory ability in small squamate reptiles. The described paradigm and procedures were successfully used with male side-blotched lizards (Uta stansburiana), demonstrating that spatial learning and memory can be assessed in this taxonomic group with an ecologically relevant apparatus and protocol.

This paper describes a modification of the Barnes maze, a standard rodent paradigm used to assess spatial memory and learning, for use in small squamate reptiles.

הערכת למידה וזיכרון המרחבי קטן קַשׂקַשִׂי זוחל

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

מבחן אובייקטיבי, לשחזור של למידה חוש הריח ואפליה בעכברים

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation

Current scientific literature provides evidence that trigeminal sensorimotor activity associated with chewing may affect arousal, attention, and cognitive performance. These effects may be due to widespread connections of the trigeminal system to the ascending reticular activating system (ARAS), to which noradrenergic neurons of the locus coeruleus (LC) belongs. LC neurons contain projections to the whole brain, and it is known that their discharge co-varies with pupil size. LC activation is necessary for eliciting task-related mydriasis. If chewing effects on cognitive performance are mediated by the LC, it is reasonable to expect that changes in cognitive performance are correlated to changes in task-related mydriasis. Two novel protocols are presented here to verify this hypothesis and document that chewing effects are not attributable to aspecific motor activation. In both protocols, performance and pupil size changes observed during specific tasks are recorded before, soon after, and half an hour following a 2 min period of either: a) no activity, b) rhythmic, bilateral handgrip, c) bilateral chewing of soft pellet, and d) bilateral chewing of hard pellet. The first protocol measures level of performance in spotting target numbers displayed within numeric matrices. Since pupil size recordings are recorded by an appropriate pupillometer that impedes vision to ensure constant illumination levels, task-related mydriasis is evaluated during a haptic task. Results from this protocol reveal that 1) chewing-induced changes in performance and task-related mydriasis are correlated and 2) neither performance nor mydriasis are enhanced by handgrip. In the second protocol, use of a wearable pupillometer allows measurement of pupil size changes and performance during the same task, thus allowing even stronger evidence to be obtained regarding LC involvement in the trigeminal effects on cognitive activity. Both protocols have been run in the historical office of Prof. Giuseppe Moruzzi, the discoverer of ARAS, at the University of Pisa.

To verify whether trigeminal effects on cognitive performance involve locus coeruleus activity, two protocols are presented that aim to evaluate possible correlations between the performance and task-related pupil size changes induced by chewing. These protocols may be applied to conditions in which locus coeruleus contribution is suspected.

הערכת שינויי האישון בליקום כפיית-עוררות בתיווך באמצעות גירוי מעורר

Current scientific literature provides evidence that trigeminal sensorimotor activity associated with chewing may affect arousal, attention, and cognitive ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

מדידה כמותית של מתוך מסונתז חלבונים בעכברים

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

Electrocorticographic Recording of Cerebral Cortex Areas Manipulated Using an Adeno-Associated Virus Targeting Cofilin in Mice

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. Adeno-associated viruses (AAVs) are increasingly used to improve understanding of brain circuits and their functions. The AAV-mediated manipulation of specific cell populations and/or of precise molecular components has been tremendously useful to identify new sleep regulatory circuits/molecules and key proteins contributing to the adverse effects of sleep loss. For instance, inhibiting activity of the filamentous actin-severing protein&#160;cofilin&#160;using AAV prevents sleep deprivation-induced memory impairment. Here, a protocol is&#160;described that combines the manipulation of cofilin function in a cerebral cortex area with the recording of ECoG activity to examine whether cortical cofilin modulates the wakefulness and sleep ECoG signals. AAV injection is performed during the same surgical procedure as the implantation of ECoG and electromyographic (EMG) electrodes in adult male and female mice. Mice are anesthetized, and their heads are shaved. After skin cleaning and incision, stereotaxic coordinates of the motor cortex are determined, and the skull is pierced at this location. A cannula prefilled with an AAV expressing cofilinS3D, an inactive form of cofilin, is slowly positioned in the cortical tissue. After AAV infusion, gold-covered screws (ECoG electrodes) are screwed through the skull and cemented to the skull with gold wires inserted in the neck muscles (EMG electrodes). The animals are allowed three weeks to recover and to ensure sufficient expression of cofilinS3D. The infected area and cell type are verified using immunohistochemistry, and the ECoG is analyzed using visual identification of vigilance states and spectral analysis. In summary, this combined methodological approach allows the investigation of the precise contribution of molecular components regulating neuronal morphology and connectivity to the regulation of synchronized cerebral cortex activity during wakefulness and sleep.

This article describes a protocol for the manipulation of molecular targets in the cerebral cortex using adeno-associated viruses and for monitoring the effects of this manipulation during wakefulness and sleep using electrocorticographic recordings.

הקלטה אלקטרוקורטיקוגרפית של אזורי קליפת המוח מניפולציה באמצעות וירוס הקשור אדנו מיקוד Cofilin בעכברים

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. ...