1) הכנה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כדי להתחיל, להפעיל את מערכת NeuroStar, להיכנס פנימה, לבצע את הבדיקה סליל אוטומטיות. </li><li style=";text-align:right;direction:rtl"> לאחר מכן, לגשת לקובץ המטופל מתאים מהרשימה זמין (כל המטופלים באמצעות מערכת NeuroStar מחובר למחשב המרכזי) </li><li style=";text-align:right;direction:rtl"> בסופו של דבר, להבטיח את ההגדרות כיסא אינם מגבילים ולהרים את זרוע כיסא כדי להקל על החולה להיכנס ולשבת. </li></ol> 2) ישיבה יישור החולה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כאשר החולה מגיע, מושב אותו / אותה בנוחות על הכיסא. </li><li style=";text-align:right;direction:rtl"> ודא הן את החולה ואת הכנס טכנאי אטמי אוזניים. </li><li style=";text-align:right;direction:rtl"> באמצעות פס דבק, להצמיד את הרצועה מיקום הראש מעל הגבות של המטופל. ודא הנקודה המרכזית ממוקמת מעל nasion. </li><li style=";text-align:right;direction:rtl"> להשעין את הכסא ולהתאים את הראש תמיכה לפי הצורך. </li><li style=";text-align:right;direction:rtl"> קלות לצרף את רצועת ראש הדף סקוטש לעיל nasion של המטופל. </li><li style=";text-align:right;direction:rtl"> הפעל את הלייזר להבטיח חוצה nasion של המטופל. </li><li style=";text-align:right;direction:rtl"> שאל את נושא לבהות בנקודה על הקיר ולהתאים את בר / P עד שהוא רמה עם העיניים. </li><li style=";text-align:right;direction:rtl"> ודא את החולה מרוכז לחלוטין להשתמש סקוטש כדי לאבטח את הראש לצד רצועות מיצוב לכיסא טיפול. </li><li style=";text-align:right;direction:rtl"> קלות להעסיק את כרית הראש לצד על מנת להבטיח יציבות גולגולתי במהלך קביעת סף המנוע. </li><li style=";text-align:right;direction:rtl"> לבסוף, להקליט את התמיכה בראש מערכת ההגדרות באמצעות ממשק מסך מגע. </li></ol> 3) קביעת סף מוטורי <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כדי לקבוע סף המנוע, להעביר את הסליל אל ראש כזה הסליל נמצא הקודקוד ואת הכרטיסייה SenStar מיושר מעל החלק העליון של האוזן של המטופל. </li><li style=";text-align:right;direction:rtl"> הזז את / בר P קדימה עד שהוא מתיישר עם הצד של הסליל. </li><li style=";text-align:right;direction:rtl"> יישור קו מרכז על סליל עם 0 לסמן את התואר על מחוון זווית סליל ולעבור בחזרה את הבר / P. </li><li style=";text-align:right;direction:rtl"> לאחר מגע ואת המיקום הוקמו, לשאול את הנושא לחוקק את המיקום טרמפיסט: זרוע כפוף 90 מעלות במרפק, אגרוף שנערך רופף, האגודל כלפי חוץ המורחבת. </li><li style=";text-align:right;direction:rtl"> בגין גירוי ולהתמקד כל תנועה של האגודל. </li><li style=";text-align:right;direction:rtl"> בהדרגה להעלות את רמת MT 0.68-1.10 עד האגודל או האצבע תואמים תנועה נתפסת. </li><li style=";text-align:right;direction:rtl"> פעם הוא הקים תנועה, להתחיל לחפש hotspot את המנוע. הזז את החוט סביב ברשת כמו דפוס, התאמת רמת MT כנדרש. המשך לחפש עד שתמצא את המיקום ואת רמת הגדרת כוח מעורר בדיוק 5 עוויתות האגודל מתוך 10 פעימות רצופות. </li><li style=";text-align:right;direction:rtl"> לאחר hotspot המנוע נקבע, הזז את / בר P קדימה עד, שוב, מתיישר עם הצד של הסליל. </li><li style=";text-align:right;direction:rtl"> השתמש ממשק מסך מגע כדי להקליט את זווית קויל / P מיקום, מיקום ה-SOA, ואת רמת צריכת חשמל. </li><li style=";text-align:right;direction:rtl"> לבסוף, לחצו על לחצן MT נמצאו. כעת אנחנו מוכנים לטיפול. </li></ol> 4) החולה טיפול <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כדי להתחיל, הזיזו את סליל 5.5 ס&quot;מ קדמית למיקום MT. זה יהיה מקום הטיפול. </li><li style=";text-align:right;direction:rtl"> ודא סליל הוא ליצור קשר עם ראשו של המטופל לשאול את החולה להישאר עדיין למשך הטיפול. </li><li style=";text-align:right;direction:rtl"> הפרמטרים הטיפול מראש לתוך המכשיר. גירוי יופק על סף מנוע 120% עם רצף הדופק של הרץ 10 למשך 4 שניות, ואחריו תקופת 26 שקט השנייה. הטיפול יימשך עבור סכום כולל של 37.5 דקות זה סך של 3,000 פעימות. </li><li style=";text-align:right;direction:rtl"> לחצו על הלחצן רצף אשר דופק ולחץ על לחצן התחל. </li><li style=";text-align:right;direction:rtl"> במהלך הטיפול, להישאר בחדר עם המטופל לבדוק מעת לעת על מנת להבטיח סליל הוא יוצר קשר עם המטופל. </li><li style=";text-align:right;direction:rtl"> לאחר רצף הדופק הושלמה, להעביר את סליל למעלה מראשו של המטופל להתרחק בראש הצד כרית תמיכה. </li><li style=";text-align:right;direction:rtl"> תרים את החלק האחורי של הכיסא הרם את היד של הכיסא. </li><li style=";text-align:right;direction:rtl"> Assist המטופל כפי שהוא / היא נעמדת. </li><li style=";text-align:right;direction:rtl"> הסר אטמי אוזניים ולגעת התנתקות כפתור לסיים את הפגישה הטיפול. </li></ol> 5) נציג תוצאות TMS מיושם בדרך כלל מדי יום במשך תקופה של 4-6 שבועות. נתונים אחרונים מראה חלק מהחולים עשויים להיות נטייה להישנות סימפטומטי לאחר 4-6 חודשים. 8 במקרה זה, המתרגלים לשקול הפעלות תחזוקה להאריך את האפקט. טיפול זה היה יעיל כ -60% מהחולים שטופלו. <img src="/files/ftp_upload/2345/2345fig1.jpg" alt="לוח 5" />

<ol>
	<li>Pascual-Leone, A., Davey, M., Wassermann, E. M., Rothwell, J., Puri, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Transcranial Magnetic Stimulation. , (2002).</li><li>Walsh, V., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Transcranial Magnetic Stimulation: A Neurochronometrics of Mind. , (2005).</li><li>Horvath, J. C., Perez, J., Forrow, L., Fregni, F., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcranial+Magnetic+Stimulation:+An+Historical+Evaluation+and+Future+Prognosis+of+Therapeutically+Relevant+Ethical+Concerns.">Transcranial Magnetic Stimulation: An Historical Evaluation and Future Prognosis of Therapeutically Relevant Ethical Concerns.</a> , .</li><li>O'Reardon, J. P., Solvason, H. B., Janicak, P. G., Sampson, S., Isenberg, K. E., Nahas, Z., McDonald, W. M., Avery, D., Fitzgerald, P. B., Loo, C., Demitrack, M. A., George, M. S., Sackheim, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+and+safety+of+transcranial+magnetic+stimulation+in+the+acute+treatment+of+major+depression:+a+multisite+randomized+control+trial.">Efficacy and safety of transcranial magnetic stimulation in the acute treatment of major depression: a multisite randomized control trial.</a> Biological Psychiatry. 62, 1208-1216 (2007).</li><li>George, M. S., Lisanby, S. H., Avery, D., McDonald, W. M., Durkalski, V., Pavlicova, M., Anderson, B., Nahas, Z., Bulow, P., Zarkowski, P., Holtzheimer, P. D., Schwartz, T., Sackheim, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+prefrontal+transcranial+magnetic+stimulation+therapy+for+major+depressive+disorder:+a+sham-controlled+randomized+study.">Daily prefrontal transcranial magnetic stimulation therapy for major depressive disorder: a sham-controlled randomized study.</a> Arch Gen Psychiatry. 67, 507-516 (2010).</li><li>Lisanbym, S. H., Husain, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+Left+Prefrontal+Repetitive+Transcranial+Magnetic+Stimulation+(rTMS)+in+the+Acute+Treatment+of+Major+Depression:+Clinical+Predictors+of+Outcome+in+a+Multisite,+Randomized+Controlled+Clinical+Trial.">Daily Left Prefrontal Repetitive Transcranial Magnetic Stimulation (rTMS) in the Acute Treatment of Major Depression: Clinical Predictors of Outcome in a Multisite, Randomized Controlled Clinical Trial.</a> Neuropsychopharmacology. 34, 522-534 (2008).</li><li>Thase, M., Demitrack, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+clinical+significance+of+treatment+outcomes+in+studies+of+resistant+major+depression.">Evaluating clinical significance of treatment outcomes in studies of resistant major depression.</a> Biological Psychiatry. 63, 138-138 (2008).</li><li>Demirtas-Tatlidede, A., Mechanic-Hamilton, D., Press, D. Z., Pearlman, C., Stern, W. M., Thall, M., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+open-label,+prospective+study+of+repetitive+transcranial+magnetic+stimulation+(rTMS)+in+the+long-term+treatment+of+refractory+depression:+reproducibility+and+duration+of+the+antidepressant+effect+in+medication-free+patients.">An open-label, prospective study of repetitive transcranial magnetic stimulation (rTMS) in the long-term treatment of refractory depression: reproducibility and duration of the antidepressant effect in medication-free patients.</a> Journal of Clinical Psychiatry. 69, 930-934 (2008).</li></ol>

הפקת וידאו של מאמר זה מומן על ידי Neuronetics, אשר מייצרת מכשירים המשמשים במחקרים אלה.

כאשר שוקלים TMS התערבות טיפולית פוטנציאל לדיכאון, חשוב לזהות את ה-FDA אישר פרוטוקול מוגדר מאוד. כדי להיות קונקרטי, אישור ה-FDA מוגבל גירוי הרץ 10 suprathreshold להחיל יומי למשך 4-6 שבועות שימוש במכשיר NeuroStar על בחולים שנכשלו להשיג שיפור משביע רצון מאחד, אבל לא יותר, מספיק טיפול תרופתי נגד דיכאון במהלך ניסויים דיכאון הנוכחית פרק. בנקודה זו, ניצול של כל דפוס גירוי חלופי, כמובן זמן חלופי, מכשיר חלופי, או האוכלוסייה המטופלת חלופי נחשב מותווים. כאשר התוצאות ממשיכות להיווצר תמיכה היעילות של טיפול TMS 5,6,7, נתונים חדשים מגלה את האפשרות של הישנות סימפטומטית מתרחש 4-6 חודשים לאחר הפסקת הטיפול. החל בפרסום זה, אין פרוטוקולים תחזוקה אושרה. לכן, נושאים הקשורים טיפול נוסף ו / או תחזוקה השפעה יש לשקול פיקוח הדוק על ידי מתרגלים בודדים.

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<td>Neuronetics Neurostar TMS Device</td>
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<td>SenStar Treatment Link</td>
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<td>Head Pad Sanitary Sheets</td>
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<td>Alignment Head Strap</td>
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the neurostar tms device: conducting the fda approved protocol for treatment of depression

NeuroStar Neuronetics Transcranial מערכת מגנטית (TMS) גירוי היא השנייה בכיתה המכשור הרפואי שמייצרת זמן קצר, פעמו שדות מגנטיים. אלו שדות במהירות לסירוגין להשרות זרמים חשמליים בתוך, אזורים נקודתיים ממוקד של קליפת המוח אשר משויכים שונים במוח שינויים פיזיולוגיים ותפקודיים. 1,2,3 בשנת 2007, O&#39;Reardon et al. ניצול המכשיר NeuroStar, פרסם את תוצאות בתעשייה בחסות, multisite, אקראי, זיוף גירוי מבוקר הניסוי הקליני שבו 301 חולים עם דיכאון מג&#39;ורי, שנכשל בעבר להגיב על אחד לפחות טיפול הולם במשפט נגד דיכאון, עבר גם TMS פעיל או אחיזת עיניים מעל הקורטקס הקדם חזיתית שמאל דורסולטרלי (DLPFC). מטופלים, שהיו תרופות ללא בזמן המחקר, קיבל TMS חמש פעמים בשבוע במשך 4-6 שבועות. 4 התוצאות הראו כי אוכלוסיית משנה של חולים (אלה שהיו יחסית פחות עמיד לתרופות, לאחר שנכשל לא יותר משני משפטים טובים תרופתי) הראה שיפור משמעותי סטטיסטית על מונטגומרי-Asberg דיכאון סולם (MADRS), את דירוג דיכאון המילטון סולם (HAMD), וכן אמצעים שונים אחרים התוצאה. בחודש אוקטובר 2008, נתמך על ידי אלה תוצאות דומות אחרות 5,6,7, השיג Neuronetics המינהל הראשונה והיחידה המזון והתרופות האמריקני (FDA) אישור לטיפול קליני של צורה מסוימת של דיכאון, תרופות עקשן באמצעות מכשיר לטיפול TMS ( ה-FDA אישור K061053). במאמר זה, נבחן שצוין FDA אישר NeuroStar פרוטוקול טיפול בדיכאון (להינתן רק תחת מרשם במקצועו רפואי מורשה או הגדרה ב-או אשפוז).

Watch this Scientific Journal Video about The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression at JoVE.com

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

במאמר זה, נבחן את המתודולוגיה שיקולים רלוונטיים פרוטוקול ה-FDA אישר טיפול בדיכאון באמצעות מכשיר NeuroStar Neuronetics TMS.

התקן TMS NeuroStar: ניהול ה-FDA פרוטוקול מאושר לטיפול דיכאון

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

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Neuroscience

39.5K Views.  Beth Israel Deaconess Medical Center. במאמר זה, נבחן את המתודולוגיה שיקולים רלוונטיים פרוטוקול ה-FDA אישר טיפול בדיכאון באמצעות מכשיר NeuroStar Neuronetics TMS.

Video: The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS: שימוש בפרוטוקול תטא-Burst לחקור Mechasnism של פלסטיות של אנשים עם תסמונת X שביר אוטיזם ו

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression (SD), as a means to develop novel therapeutic targets for episodic and chronic migraine. SD is the likely cause of migraine aura and migraine pain. It is a paroxysmal loss of neuronal function triggered by initially increased neuronal activity, which slowly propagates within susceptible brain regions. Normal brain function is exquisitely sensitive to, and relies on, coincident low-level immune signaling. Thus, neural immune signaling likely affects electrical activity of SD, and therefore migraine. Pain perception studies of SD in whole animals are fraught with difficulties, but whole animals are well suited to examine systems biology aspects of migraine since SD activates trigeminal nociceptive pathways. However, whole animal studies alone cannot be used to decipher the cellular and neural circuit mechanisms of SD. Instead, in vitro preparations where environmental conditions can be controlled are necessary. Here, it is important to recognize limitations of acute slices and distinct advantages of hippocampal slice cultures. Acute brain slices cannot reveal subtle changes in immune signaling since preparing the slices alone triggers: pro-inflammatory changes that last days, epileptiform behavior due to high levels of oxygen tension needed to vitalize the slices, and irreversible cell injury at anoxic slice centers. 
In contrast, we examine immune signaling in mature hippocampal slice cultures since the cultures closely parallel their in vivo counterpart with mature trisynaptic function; show quiescent astrocytes, microglia, and cytokine levels; and SD is easily induced in an unanesthetized preparation. Furthermore, the slices are long-lived and SD can be induced on consecutive days without injury, making this preparation the sole means to-date capable of modeling the neuroimmune consequences of chronic SD, and thus perhaps chronic migraine. We use electrophysiological techniques and non-invasive imaging to measure neuronal cell and circuit functions coincident with SD. Neural immune gene expression variables are measured with qPCR screening, qPCR arrays, and, importantly, use of cDNA preamplification for detection of ultra-low level targets such as interferon-gamma using whole, regional, or specific cell enhanced (via laser dissection microscopy) sampling. Cytokine cascade signaling is further assessed with multiplexed phosphoprotein related targets with gene expression and phosphoprotein changes confirmed via cell-specific immunostaining. Pharmacological and siRNA strategies are used to mimic and modulate SD immune signaling.

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Migraine and its transformation to chronic migraine are immense healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression in hippocampal slice cultures, as a means to develop novel therapeutic targets.

דוגמנות איתות החיסון עצבית של מיגרנה אפיזודי כרוני באמצעות הפצת דיכאון במבחנה</em

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options.  We seek to define how neural immune ...

A Comprehensive Protocol for Manual Segmentation of the Medial Temporal Lobe Structures

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, hippocampus, and the associated parahippocampal regions (perirhinal, entorhinal,&#160;and parahippocampal proper). Unlike most other tracing protocols available, typically focusing on certain MTL areas (e.g., amygdala and/or hippocampus), the integrative perspective adopted by the present tracing guidelines allows for clear localization of all MTL subregions. By integrating information from a variety of sources, including extant tracing protocols separately targeting various MTL structures, histological reports, and brain atlases, and with the complement of illustrative visual materials, the present protocol provides an accurate, intuitive, and convenient guide for understanding the MTL anatomy. The need for such tracing guidelines is also emphasized by illustrating possible differences between automatic and manual segmentation protocols. This knowledge can be applied toward research involving not only structural MRI investigations but also structural-functional colocalization and fMRI signal extraction from anatomically defined ROIs, in healthy and clinical groups alike.

The present work provides a comprehensive set of guidelines for manually tracing the medial temporal lobe (MTL) structures. This protocol can be applied to research involving structural and/or combined structural-functional magnetic resonance imaging (MRI) investigations of the MTL, in both healthy and clinical groups.

מקיף פרוטוקול לפילוח ידני של מבנים המדיאלי Temporal האונה

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, ...

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol (TRAP) Assay

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and lifespan. In addition, telomerase protects the mitochondria from oxidative stress and confers resistance to apoptosis, suggesting its possible importance for the surviving of non-mitotic, highly active cells such as neurons. We previously demonstrated the ability of novel telomerase activators to increase telomerase activity and expression in the various mouse brain regions and to protect motor neurons cells from oxidative stress. These results strengthen the notion that telomerase is involved in the protection of neurons from various lesions. To underline the role of telomerase in the brain, we here compare the activity of telomerase in male and female mouse brain and its dependence on age. TRAP assay is a standard method for detecting telomerase activity in various tissues or cell lines. Here we demonstrate the analysis of telomerase activity in three regions of the mouse brain by non-denaturing protein extraction using CHAPS lysis buffer followed by modification of the standard TRAP assay.
In this 2-step assay, endogenous telomerase elongates a specific telomerase substrate (TS primer) by adding TTAGGG 6 bp repeats (telomerase reaction). The telomerase reaction products are amplified by PCR reaction creating a DNA ladder of 6 bp increments. The analysis of the DNA ladder is made by 4.5% high resolution agarose gel electrophoresis followed by staining with highly sensitive nucleic acid stain.
Compared to the traditional TRAP assay that utilize 32P labeled radioactive dCTP&#39;s for DNA detection and polyacrylamide gel electrophoresis for resolving the DNA ladder, this protocol offers a non-toxic time saving TRAP assay for evaluating telomerase activity in the mouse brain, demonstrating the ability to detect differences in telomerase activity in the various female and male mouse brain region.

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol TRAP Assay

Telomerase is expressed in the neonatal brain and also in distinct regions of the adult brain. We present a non-toxic time saving TRAP assay for the analysis of telomerase activity in various regions of the mouse brain and detection of differences in telomerase activity between male and female mouse brains.

פעילות telomerase באזורים שונים של מוח עכבר: הטלומרז לא רדיואקטיבי חזור הגברה פרוטוקול Assay (TRAP)

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and ...

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have validated the utility of this method for enhancing neuronal preservation and overall brain slice viability. The implementation of this technique by early adopters has facilitated detailed investigations into brain function using diverse experimental applications and spanning a wide range of animal ages, brain regions, and cell types. Steps are outlined for carrying out the protective recovery brain slice technique using an optimized NMDG artificial cerebrospinal fluid (aCSF) media formulation and enhanced procedure to reliably obtain healthy brain slices for patch clamp electrophysiology. With this updated approach, a substantial improvement is observed in the speed and reliability of gigaohm seal formation during targeted patch clamp recording experiments while maintaining excellent neuronal preservation, thereby facilitating challenging experimental applications. Representative results are provided from multi-neuron patch clamp recording experiments to assay synaptic connectivity in neocortical brain slices prepared from young adult transgenic mice and mature adult human neurosurgical specimens. Furthermore, the optimized NMDG protective recovery method of brain slicing is compatible with both juvenile and adult animals, thus resolving a limitation of the original methodology. In summary, a single media formulation and brain slicing procedure can be implemented across various species and ages to achieve excellent viability and tissue preservation.

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol demonstrates the implementation of an optimized N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. A single media formulation is used to reliably obtain healthy brain slices from animals of any age and for diverse experimental applications.

הכנת פרוסות המוח חריפה באמצעות אופטימיזציה N-מתיל-D-שיטת שחזור מגן glucamine

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have ...

Receptor Autoradiography Protocol for the Localized Visualization of Angiotensin II Receptors

This protocol describes receptor binding patterns for Angiotensin II (Ang II) in the rat brain using a radioligand specific for Ang II receptors to perform receptor autoradiographic mapping.
Tissue specimens are harvested and stored at -80 &#176;C. A cryostat is used to coronally section the tissue (brain) and thaw-mount the sections onto charged slides. The slide-mounted tissue sections are incubated in 125I-SI-Ang II to radiolabel Ang II receptors. Adjacent slides are separated into two sets: &#39;non-specific binding&#39; (NSP) in the presence of a receptor saturating concentration of non-radiolabeled Ang II, or an AT1 Ang II receptor subtype (AT1R) selective Ang II receptor antagonist, and &#39;total binding&#39; with no AT1R antagonist. A saturating concentration of AT2 Ang II receptor subtype (AT2R) antagonist (PD123319, 10 &#181;M) is also present in the incubation buffer to limit 125I-SI-Ang II binding to the AT1R subtype. During a 30 min pre-incubation at ~22 &#176;C, NSP slides are exposed to 10 &#181;M PD123319 and losartan, while &#39;total binding&#39; slides are exposed to 10 &#181;M PD123319. Slides are then incubated with 125I-SI-Ang II in the presence of PD123319 for &#39;total binding&#39;, and PD123319 and losartan for NSP in assay buffer, followed by several &#39;washes&#39; in buffer, and water to remove salt and non-specifically bound radioligand. The slides are dried using blow-dryers, then exposed to autoradiography film using a specialized film and cassette. The film is developed and the images are scanned into a computer for visual and quantitative densitometry using a proprietary imaging system and a spreadsheet. An additional set of slides are thionin-stained for histological comparisons.
The advantage of using receptor autoradiography is the ability to visualize Ang II receptors in situ, within a section of a tissue specimen, and anatomically identify the region of the tissue by comparing it to an adjacent histological reference section.

Here we present a protocol to describe the localization of angiotensin II Type 1 receptors in the rat brain by quantitative, densitometric, in vitro receptor autoradiography using an iodine-125 labeled analog of angiotensin II.

קולטן autoradiography פרוטוקול עבור ויזואליזציה המרוכזת של אנגיוטנסין II רצפטורים

This protocol describes receptor binding patterns for Angiotensin II (Ang II) in the rat brain using a radioligand specific for Ang II receptors to ...

Protocol for Isolating the Mouse Circle of Willis

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and hypothalamus that supplies blood to the brain and surrounding structures. It has been implicated in several cerebrovascular disorders, including cerebral amyloid angiopathy (CAA)-associated vasculopathies, intracranial atherosclerosis and intracranial aneurysms. Studies of the molecular mechanisms underlying these diseases for the identification of novel drug targets for their prevention require animal models. Some of these models may be transgenic, whereas others will involve isolation of the cerebro-vasculature, including the CoW.The method described here is suitable for CoW isolation in any mouse lineage and has considerable potential for screening (expression of genes, protein production, posttranslational protein modifications, secretome analysis, etc.) studies on the large vessels of the mouse cerebro-vasculature. It can also be used for ex vivo studies, by adapting the organ bath system developed for isolated mouse olfactory arteries.

We describe here a reproducible protocol for isolating the mouse circle of Willis.

פרוטוקול בידוד לחוג עכבר של וויליס

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

צעד-אחד פשוט פרוטוקול לנתיחה עבור כל הר הכנה למבוגרים<em&gt; תסיסנית</em&gt; Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

A Protocol for the Administration of Real-Time fMRI Neurofeedback Training

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control neuroplastic processes and correct abnormal function, or even shift functions from damaged tissue to physiologically healthy brain regions, hold the potential to dramatically improve overall health. Of the current neuroplastic interventions in development, neurofeedback training (NFT) from functional Magnetic Resonance Imaging (fMRI) has the advantages of being completely non-invasive, non-pharmacologic, and spatially localized to target brain regions, as well as having no known side effects. Furthermore, NFT techniques, initially developed using fMRI, can often be translated to exercises that can be performed outside of the scanner without the aid of medical professionals or sophisticated medical equipment. In fMRI NFT, the fMRI signal is measured from specific regions of the brain, processed, and presented to the participant in real-time. Through training, self-directed mental processing techniques, that regulate this signal and its underlying neurophysiologic correlates, are developed. FMRI NFT has been used to train volitional control over a wide range of brain regions with implications for several different cognitive, behavioral, and motor systems. Additionally, fMRI NFT has shown promise in a broad range of applications such as the treatment of neurologic disorders and the augmentation of baseline human performance. In this article, we present an fMRI NFT protocol developed at our institution for modulation of both healthy and abnormal brain function, as well as examples of using the method to target both cognitive and auditory regions of the brain.

The ability to induce and/or control neural plasticity may be critical in future treatments for neurologic disorders and the recovery from brain injury. In this paper, we present a protocol on the use of neurofeedback training with functional magnetic resonance imaging to modulate human brain function.

פרוטוקול עבור fMRI ניהול בזמן אמת Neurofeedback אימונים

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control ...

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the influence of mechanical stress on cellular function is still poorly understood. In part, this knowledge gap exists because few tools enable simultaneous deformation of tissue and cells, imaging of cellular activity in live animals, and efficient restriction of motility in otherwise highly mobile model organisms, such as the nematode Caenorhabditis elegans. The small size of C. elegans makes them an excellent match to microfluidics-based research devices, and solutions for immobilization have been presented using microfluidic devices. Although these devices allow for high-resolution imaging, the animal is fully encased in polydimethylsiloxane (PDMS) and glass, limiting physical access for delivery of mechanical force or electrophysiological recordings. Recently, we created a device that integrates pneumatic actuators with a trapping design that is compatible with high-resolution fluorescence microscopy. The actuation channel is separated from the worm-trapping channel by a thin PDMS diaphragm. This diaphragm is deflected into the side of a worm by applying pressure from an external source. The device can target individual mechanosensitive neurons. The activation of these neurons is imaged at high-resolution with genetically-encoded calcium indicators. This article presents the general method using C. elegans strains expressing calcium-sensitive activity indicator (GCaMP6s) in their touch receptor neurons (TRNs). The method, however, is not limited to TRNs nor to calcium sensors as a probe, but can be expanded to other mechanically-sensitive cells or sensors.

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

New tools for mechanobiology research are needed to understand how mechanical stress activates biochemical pathways and elicits biological responses. Here, we showcase a new method for selective mechanical stimulation of immobilized animals with a microfluidic trap allowing high-resolution imaging of cellular responses.