1. קביעת סף מוטורי מנוחה: <ol class="lalpha" style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כדי להתחיל, מושב בנושא שלך על כיסא נוח. </li><li style=";text-align:right;direction:rtl"> הבא, להחיל אלקטרודות EMG על פני שריר inerosseus הראשון הגבי של יד ימין של הנושא. (INSERT FIGURE1) אם אתה מתקשה באיתור השריר, לשאול את הנושא כדי ליצור תנועה צובט או להעביר שלו / שלה האצבע הלוך ושוב - זה צריך לעשות שריר קל יותר לאתר. </li><li style=";text-align:right;direction:rtl"> מניחים אלקטרודה הקרקע על החלק הבשרני של האמה של הנושא. </li><li style=";text-align:right;direction:rtl"> קבע את המיקום של הקורטקס המוטורי שמאל של הנבדק. אמנם אזור זה שונה במקצת עבור כולם, זה יהיה בדרך כלל להיות סביב 3.5 ס&quot;מ rostrolateral של קודקוד הגולגולת. אל תפחד לחקור רשת קטן סביב האזור הזה ולהתאים את זווית גירוי עד למצוא את המנוע &quot;נקודה חמה&quot;. </li><li style=";text-align:right;direction:rtl"> לאחר שמצאת M1, החל כוח של 50%, להתחיל גירוי הדופק יחיד באזור ולמדוד את חברי הפרלמנט האירופי עורר כתוצאה. </li><li style=";text-align:right;direction:rtl"> כוון את העוצמה למעלה או למטה עד שאתה מסוגל לייצר MEP μV של יותר מ 50 בדיוק ב 5 של 10 פעימות רצופות. רמה זו היא סף כוח המנוע מנוחה של הנושא. </li></ol> 2. קביעת סף מנוע פעיל <ol class="lalpha" style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> באמצעות אלקטרודה אותו הגדרת כמתואר לעיל, לשאול את נושא לצבוט שלו / שלה לאצבע אגודל ימין יחד באמצעות לחץ על 20%. </li><li style=";text-align:right;direction:rtl"> בעוד הנושא מחזיק בעמדה זו צובט, להתחיל הדופק יחיד גירוי בהספק 50% מעל M1 ולמדוד את חברי הפרלמנט האירופי שנוצר. </li><li style=";text-align:right;direction:rtl"> כוון את העוצמה למעלה או למטה עד שאתה מסוגל לייצר MEP μV של יותר מ 200 על בדיוק 5 של 10 פעימות רצופות. רמה זו היא סף כוח המנוע פעיל של הנושא. </li></ol> 3. קבע Baseline MEP <ol class="lalpha" style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כדי לקבוע בסיס MEP של נושא משרעת, לעורר המנוע חמה על סף 120% מנוע מנוחה עם 10 פולסים יחיד. </li><li style=";text-align:right;direction:rtl"> מדוד את חבר הפרלמנט האירופי, אחרי הדופק בכל. </li><li style=";text-align:right;direction:rtl"> ממוצע MEP הערכים משלוש קבוצות של 10 פעימות. </li></ol> 4. הגדרת פרמטרים TMS <ol class="lalpha" style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> אנחנו נהיה ניצול תקן תטא-Burst פרמטרים: שלושה פולסים בתדר 50 הרץ בעצימות של 80% סף פעיל מנוע עבור 200 מילישניות במרווחים. </li><li style=";text-align:right;direction:rtl"> כדי לנהל לסירוגין כפות, ליצור רכבת של שתי שניות סטים (בכל הפרמטרים הנ&quot;ל) ואחריו שמונה שניות של גירוי לא. חזור עבור 190 שניות (סך של 600 פולסים). iTBS הוכח כדי להקל על פעילות קליפת המוח. </li><li style=";text-align:right;direction:rtl"> כדי לנהל רציף תטא-Burst, ליצור רכבת סטים רצוף (על הפרמטרים הנ&quot;ל) במשך 47 שניות (סך של 600 פולסים). cTBS הוכח לעכב פעילות קליפת המוח. </li></ol> 5. TMS גירוי <ol class="lalpha" style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> עבור פרוטוקול זה, לנהל או iTBS או cTBS על hotspot הוקמה קודם לכן מנוע M1. </li></ol> 6. קבע פוסט גירוי MEP <ol class="lalpha" style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> בעקבות TBS, כמו קודם, לעורר המנוע חמה על סף 120% מנוע מנוחה עם 10 פולסים יחיד. </li><li style=";text-align:right;direction:rtl"> אצווה ממוצע של 10 רמות MEP. </li><li style=";text-align:right;direction:rtl"> איסוף חדשים 10-דופק ממוצע אצווה ב 5 דקות, 10 דקות, 20, 30, 40, 50, 60, 75, 90, 105, ו 120 דקות לאחר גירוי. </li></ol> 7. נציג תוצאות כאשר ניצול דפוס iTBS, כדאי לשים לב לעליה ערכים MEP למשך המשקף המצב הטבעי של הנושא של פלסטיות. כמו כן, כאשר ניצול דפוס cTBS, אתה צריך לשים לב לירידה ערכים MEP למשך המשקף המצב הטבעי של הנושא של פלסטיות. <img src="/files/ftp_upload/2272/2272fig1.jpg" alt="איור 1" /> באיור 1. תרשים של מיקום האלקטרודה למדידת MEP. <img src="/files/ftp_upload/2272/2272fig2.jpg" alt="איור 2" /> איור 2. דיאגרמה של דפוס גירוי iTBS. <img src="/files/ftp_upload/2272/2272fig3.jpg" alt="איור 3" /> איור 3. דיאגרמה של דפוס גירוי cTBS.

<ol>
	<li>Pfeiffer, B. E., Huber, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+state+of+synapses+in+fragile+x+syndrome.">The state of synapses in fragile x syndrome.</a> Neuroscientist. 15, 549-567 (2009).</li><li>O'Donnel, W. T., Warren, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+decade+of+molecular+studies+of+fragile+x+syndrome.">A decade of molecular studies of fragile x syndrome.</a> Annual Review of Neuroscience. 25, 315-338 (2002).</li><li>Tassone, F., Hagerman, R. J., Taylor, A. K., Mills, J. B., Wood, S., Gane, L. W., Hagerman, P. 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> American Journal of Human Genetics. 65, A493-A493 (1999).</li><li>Hagerman, R. J., Miller, L. J., McGrath-Clarke, J., Riley, K., Goldson, E., Harris, S. W., Simon, J., Church, K., Bonnell, J., Ognibene, T. C., McIntosh, D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+stimulants+on+electrodermal+studies+in+fragile+x+syndrome.">Influence of stimulants on electrodermal studies in fragile x syndrome.</a> Microscopy Research and Technique. 57, 168-173 (2002).</li><li>Miller, L. J., McIntosh, D. N., McGrath, J., Shyu, V., Lampe, M., Taylor, A. K., Tassone, F., Neitzel, K., Stackhouse, T., Hagerman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrodermal+response+to+sensory+stimuli+in+individuals+with+fragile+x+syndrome:+a+preliminary+report.">Electrodermal response to sensory stimuli in individuals with fragile x syndrome: a preliminary report.</a> American Journal of Medical Genetics. 83, 268-279 (1999).</li><li>Hagerman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fragile+X:+perspectives+from+birth+through+aging.">Fragile X: perspectives from birth through aging.</a> Biological Psychiatry. 57, 73s-73s (2005).</li><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>Thickbroom, G. W. <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+and+synaptic+plasticity:+experimental+framework+and+human+models.">Transcranial magnetic stimulation and synaptic plasticity: experimental framework and human models.</a> Experimental Brain Research. 180, 583-593 (2007).</li><li>Huang, Y. Z., Edwards, M. J., Rounis, E., Bhatia, K. P., Rothwell, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theta+burst+stimulation+of+the+human+motor+cortex.">Theta burst stimulation of the human motor cortex.</a> Neuron. 45, 201-206 (2005).</li><li>Oberman, L., Eldaief, M. C., Gautam, S., Fecteau, S., Tormos, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Abnormal+Cortical+Plasticity+in+Adults+with+Asperger's+Syndrome.">Abnormal Cortical Plasticity in Adults with Asperger's Syndrome.</a> , .</li></ol>

אין ניגודי אינטרסים הכריז.

בידיעה כי משך הזמן הממושך של TMS לאחר לוואי הוא שיקוף של פלסטיות עצבית טיפוסיות, מחקרים כגון זה יכול לשמש כדי ליצור פרוטוקולים אבחון טוב יותר ומדויק יותר האוטיזם והפרעות הקשורות. בדומה לכך, עם נתונים נוספים, נוכל למצוא סמנים ביולוג...

<table><tbody><tr><td>Alcohol Swabs</td><td></td><td></td><td></td></tr><tr><td>EMG Electrodes</td><td></td><td></td><td></td></tr><tr><td>EMG Measuring Equipment/Software</td><td></td><td></td><td></td></tr><tr><td>Ear Plugs</td><td></td><td></td><td></td></tr><tr><td>Marker or Grease Pen</td><td></td><td></td><td></td></tr><tr><td>Any Single Pulse Capable TMS Device</td><td></td><td></td><td></td></tr><tr><td>Any rTMS Capable TMS Device (for TBS)</td><td></td><td></td><td></td></tr><tr><td>Any Compatible Figure-of-Eight Coil</td><td></td><td></td><td></td></tr><tr><td>Frameless Stereotactic Equipment (Optional)</td><td></td><td></td><td></td></tr></tbody></table>

tms: using the theta-burst protocol to explore mechanism of plasticity in individuals with fragile x syndrome and autism

תסמונת X שביר (FXS), הידוע גם בשם תסמונת מרטין בל, היא פגם גנטי שנמצא על כרומוזום X 1,2 אנשים הסובלים הפרעות FXS להציג בביטוי של FMR1 -. חלבון הדרוש לפיתוח, טיפוסי עצביים בריאה. 3 נתונים אחרונים הראו כי אובדן של חלבון זה יכול לגרום בקליפת להיות hyperexcitable ובכך להשפיע על דפוסי הכוללת של פלסטיות עצבית. 4,5 בנוסף, X שביר מראה תחלואה נלווית חזק עם אוטיזם: למעשה, 30% מהילדים עם FXS מאובחנים עם אוטיזם, ו 2 - 5% של ילדים אוטיסטים סובלים FXS 6. גירוי מגנטי Transcranial (א neurostimulatory פולשני וטכניקה neuromodulatory שיכולים transiently לאורך זמן רב או לווסת את רגישות קליפת המוח באמצעות היישום של שדה מגנטי מקומי פולסים 7,8) מייצג שיטה ייחודית של חקר הפלסטיות לבין גילויי FXS בתוך אנשים מושפעים. באופן ספציפי יותר, תטא-Burst גירוי (TBS), פרוטוקול גירוי ספציפי הראו לווסת פלסטיות בקליפת המוח למשך עד 30 דקות לאחר הפסקת הגירוי באוכלוסיות בריא, כבר הוכיחה את כלי יעיל בחקר של פלסטיות נורמלי. 9,10 מחקרים שנעשו לאחרונה הראו את ההשפעות של כפות האחרון ניכרת יותר אצל אנשים על הספקטרום האוטיסטי -. עד 90 דקות 11 זה המורחבת השפעה, משך מציע חריגות היסודי במצב פלסטיות הטבעית של המוח אצל אנשים עם אוטיזם - בדומה לרגשנות יתר הנגרמת על ידי שביר תסמונת X. בניסוי זה, ניצול יחיד הדופק מנוע עורר הפוטנציאלים (חברי הפרלמנט האירופי) כאמת מידה שלנו, נוכל לחקור את ההשפעות של כפות שניהם לסירוגין רציפה על הפלסטיות קליפת המוח אצל אנשים הסובלים FXS ויחידים על הספקטרום האוטיסטי.

Watch this Scientific Journal Video about TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism at JoVE.com

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

במאמר זה, נבחן את ההשפעות של גירוי TMS תטא-Burst על הפלסטיות קליפת המוח אצל אנשים הסובלים מתסמונת ה-X השביר ויחידים על הספקטרום האוטיסטי.

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 ...

tms-using-theta-burst-protocol-to-explore-mechanism-plasticity

Research

JoVE Journal

Neuroscience

16.8K Views.  Beth Israel Deaconess Medical Center. במאמר זה, נבחן את ההשפעות של גירוי TMS תטא-Burst על הפלסטיות קליפת המוח אצל אנשים הסובלים מתסמונת ה-X השביר ויחידים על הספקטרום האוטיסטי.

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

Probing the Brain in Autism Using fMRI and Diffusion Tensor Imaging

Newly emerging theories suggest that the brain does not function as a cohesive unit in autism, and this discordance is reflected in the behavioral symptoms displayed by individuals with autism. While structural neuroimaging findings have provided some insights into brain abnormalities in autism, the consistency of such findings is questionable. Functional neuroimaging, on the other hand, has been more fruitful in this regard because autism is a disorder of dynamic processing and allows examination of communication between cortical networks, which appears to be where the underlying problem occurs in autism. Functional connectivity is defined as the temporal correlation of spatially separate neurological events1. Findings from a number of recent fMRI studies have supported the idea that there is weaker coordination between different parts of the brain that should be working together to accomplish complex social or language problems2,3,4,5,6. One of the mysteries of autism is the coexistence of deficits in several domains along with relatively intact, sometimes enhanced, abilities. Such complex manifestation of autism calls for a global and comprehensive examination of the disorder at the neural level. A compelling recent account of the brain functioning in autism, the cortical underconnectivity theory,2,7 provides an integrating framework for the neurobiological bases of autism. The cortical underconnectivity theory of autism suggests that any language, social, or psychological function that is dependent on the integration of multiple brain regions is susceptible to disruption as the processing demand increases. In autism, the underfunctioning of integrative circuitry in the brain may cause widespread underconnectivity. In other words, people with autism may interpret information in a piecemeal fashion at the expense of the whole. Since cortical underconnectivity among brain regions, especially the frontal cortex and more posterior areas 3,6, has now been relatively well established, we can begin to further understand brain connectivity as a critical component of autism symptomatology.
A logical next step in this direction is to examine the anatomical connections that may mediate the functional connections mentioned above. Diffusion Tensor Imaging (DTI) is a relatively novel neuroimaging technique that helps probe the diffusion of water in the brain to infer the integrity of white matter fibers. In this technique, water diffusion in the brain is examined in several directions using diffusion gradients. While functional connectivity provides information about the synchronization of brain activation across different brain areas during a task or during rest, DTI helps in understanding the underlying axonal organization which may facilitate the cross-talk among brain areas. This paper will describe these techniques as valuable tools in understanding the brain in autism and the challenges involved in this line of research.

Neuroimaging techniques, such as functional MRI and Diffusion Tensor Imaging have become increasingly useful in characterizing the cognitive and neural deficits in autism. An examination of brain connectivity in autism at a network level along with adaptations for scanning children with developmental disabilities is presented.

לחקור את המוח באוטיזם באמצעות fMRI ו דימות מותח דיפוזי

Newly emerging theories suggest that the brain does not function as a cohesive unit in autism, and this discordance is reflected in the behavioral ...

Medicine

A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene

Fragile X syndrome (FXS) and associated disorders are caused by expansion of the cytosine-guanine-guanine (CGG) trinucleotide repeat in the 5&#8217; untranslated region (UTR) of the Fragile X mental retardation-1 (FMR1) gene promoter. Conventionally, capillary electrophoresis fragment analysis on a genetic analyzer is used for the sizing of the CGG repeats of FMR1, but additional Southern blot analysis is required for exact measurement when the repeat number is higher than 200. Here, we present an accurate and robust polymerase chain reaction (PCR)-based method for quantification of the CGG repeats of FMR1. The first step of this test is PCR amplification of the repeat sequences in the 5&#8217;UTR of the FMR1 promoter using a Fragile X PCR kit, followed by purification of the PCR products and fragment sizing on a microfluidic capillary electrophoresis instrument, and subsequent interpretation of the number of CGG repeats by referencing standards with known repeats using the analysis software. This PCR-based assay is reproducible and capable of identifying the full range of CGG repeats of FMR1 promoters, including those with a repeat number of more than 200 (classified as full mutation), 55 to 200 (premutation), 46 to 54 (intermediate), and 10 to 45 (normal). It is a cost-effective method that facilitates classification of the FXS and Fragile X-associated disorders with robustness and rapid reporting time.

An accurate and robust polymerase chain reaction-based assay for quantifying cytosine-guanine-guanine trinucleotide repeats in the Fragile X mental retardation-1 gene facilitates molecular diagnosis and screening of Fragile X syndrome and Fragile X-related disorders with shorter turn-around time and investment in equipment.

בסיס מבוסס על תגובת שרשרת פולימראז חזקה עבור כימות ציטוסינוס-גואנין-גואנין Trinucleotide גאות חוזר בצורה שברירית X פיגור-1 גן

Fragile X syndrome (FXS) and associated disorders are caused by expansion of the cytosine-guanine-guanine (CGG) trinucleotide repeat in the 5&#8217; ...

Genetics

An Electrophysiology Protocol to Measure Reward Anticipation and Processing in Children

We present a protocol designed to measure the neural correlates of reward in children. The protocol allows researchers to measure both reward anticipation and processing. Its purpose is to create a reward task that is appropriate for young children with and without autism while controlling reward properties between two conditions: social and nonsocial. The current protocol allows for comparisons of brain activity between social and nonsocial reward conditions while keeping the reward itself identical between conditions. Using this protocol, we found evidence that neurotypical children demonstrate enhanced anticipatory brain activity during the social condition. Furthermore, we found that neurotypical children anticipate social reward more robustly than children with autism diagnoses. As the task uses snacks as a reward, it is most appropriate for young children. However, the protocol may be adapted for use with adolescent or adult populations if snacks are replaced by monetary incentives. The protocol is designed to measure electrophysiological events (event-related potentials), but it may be customized for use with eye-tracking or fMRI.

This protocol is designed to measure reward anticipation and processing in young children with and without autism. Specifically, the protocol is designed to study the neural correlates of reward during social and nonsocial conditions while controlling for reward between conditions.

פרוטוקול אלקטרופיזיולוגיה אמצעי תגמול ציפייה, עיבוד אצל ילדים

We present a protocol designed to measure the neural correlates of reward in children. The protocol allows researchers to measure both reward anticipation ...

Behavior

Rapid Detection of Neurodevelopmental Phenotypes in Human Neural Precursor Cells (NPCs)

Human brain development proceeds through a series of precisely orchestrated processes, with earlier stages distinguished by proliferation, migration, and neurite outgrowth; and later stages characterized by axon/dendrite outgrowth and synapse formation. In neurodevelopmental disorders, often one or more of these processes are disrupted, leading to abnormalities in brain formation and function. With the advent of human induced pluripotent stem cell (hiPSC) technology, researchers now have an abundant supply of human cells that can be differentiated into virtually any cell type, including neurons. These cells can be used to study both normal brain development and disease pathogenesis. A number of protocols using hiPSCs to model neuropsychiatric disease use terminally differentiated neurons or use 3D culture systems termed organoids. While these methods have proven invaluable in studying human disease pathogenesis, there are some drawbacks. Differentiation of hiPSCs into neurons and generation of organoids are lengthy and costly processes that can impact the number of experiments and variables that can be assessed. In addition, while post-mitotic neurons and organoids allow the study of disease-related processes, including dendrite outgrowth and synaptogenesis, they preclude the study of earlier processes like proliferation and migration. In neurodevelopmental disorders, such as autism, abundant genetic and post-mortem evidence indicates defects in early developmental processes. Neural precursor cells (NPCs), a highly proliferative cell population, may be a suitable model in which to ask questions about ontogenetic processes and disease initiation. We now extend methodologies learned from studying development in mouse and rat cortical cultures to human NPCs. The use of NPCs allows us to investigate disease-related phenotypes and define how different variables (e.g., growth factors, drugs) impact developmental processes including proliferation, migration, and differentiation in only a few days. Ultimately, this toolset can be used in a reproducible and high-throughput manner to identify disease-specific mechanisms and phenotypes in neurodevelopmental disorders.

Rapid Detection of Neurodevelopmental Phenotypes in Human Neural Precursor Cells NPCs

Neurodevelopmental processes such as proliferation, migration, and neurite outgrowth are often perturbed in neuropsychiatric diseases. Thus, we present protocols to rapidly and reproducibly assess these neurodevelopmental processes in human iPSC-derived NPCs. These protocols also allow the assessment of the effects of relevant growth factors and therapeutics on NPC development.

זיהוי מהיר של פנוטיפים התפתחותיות בתאי האדם קודמן עצבית (NPCs)

Human brain development proceeds through a series of precisely orchestrated processes, with earlier stages distinguished by proliferation, migration, and ...

Developmental Biology

Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological impairments 1. Two pervasive and disabling symptoms experienced by TBI survivors, memory deficits and a reduction in seizure threshold, are thought to be mediated by TBI-induced hippocampal dysfunction 2,3. In order to demonstrate how altered hippocampal circuit function adversely affects behavior after TBI in mice, we employ lateral fluid percussion injury, a commonly used animal model of TBI that recreates many features of human TBI including neuronal cell loss, gliosis, and ionic perturbation 4-6.
	Here we demonstrate a combinatorial method for investigating TBI-induced hippocampal dysfunction. Our approach incorporates multiple ex vivo physiological techniques together with animal behavior and biochemical analysis, in order to analyze post-TBI changes in the hippocampus. We begin with the experimental injury paradigm along with behavioral analysis to assess cognitive disability following TBI. Next, we feature three distinct ex vivo recording techniques: extracellular field potential recording, visualized whole-cell patch-clamping, and voltage sensitive dye recording. Finally, we demonstrate a method for regionally dissecting subregions of the hippocampus that can be useful for detailed analysis of neurochemical and metabolic alterations post-TBI.
	These methods have been used to examine the alterations in hippocampal circuitry following TBI and to probe the opposing changes in network circuit function that occur in the dentate gyrus and CA1 subregions of the hippocampus (see Figure 1). The ability to analyze the post-TBI changes in each subregion is essential to understanding the underlying mechanisms contributing to TBI-induced behavioral and cognitive deficits. 
	The multi-faceted system outlined here allows investigators to push past characterization of phenomenology induced by a disease state (in this case TBI) and determine the mechanisms responsible for the observed pathology associated with TBI.

A multi-faceted approach to investigating functional changes to hippocampal circuitry is explained. Electrophysiological techniques are described along with the injury protocol, behavioral testing and regional dissection method. The combination of these techniques can be applied in similar fashion for other brain regions and scientific questions.

חקירות על שינויים בתפקוד המעגל היפוקמפוס בעקבות פגיעה מוחית טראומטית קלה

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological ...

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct electrophysiological characterization of the synaptic connections between individual neurons, but also pharmacological manipulation of either the presynaptic or the postsynaptic neuron. When carried out in organotypic hippocampal slice cultures, the probability that two neurons are synaptically connected is significantly increased. This preparation readily enables identification of cell types, and the neurons maintain their morphology and properties of synaptic function similar to that in native brain tissue. A major advantage of paired whole cell recordings is the highly precise information it can provide on the properties of synaptic transmission and plasticity that are not possible with other more crude techniques utilizing extracellular axonal stimulation. Paired whole cell recordings are often perceived as too challenging to perform. While there are challenging aspects to this technique, paired recordings can be performed by anyone trained in whole cell patch clamping provided specific hardware and methodological criteria are followed. The probability of attaining synaptically connected paired recordings significantly increases with healthy organotypic slices and stable micromanipulation allowing independent attainment of pre- and postsynaptic whole cell recordings. While CA3-CA3 pyramidal cell pairs are most widely used in the organotypic slice hippocampal preparation, this technique has also been successful in CA3-CA1 pairs and can be adapted to any neurons that are synaptically connected in the same slice preparation. In this manuscript we provide the detailed methodology and requirements for establishing this technique in any laboratory equipped for electrophysiology.

Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.

הקלטות תא כל לזווג בפרוסות בהיפוקמפוס Organotypic

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct ...

<meta charset="utf8"></head><body>מבחנים התנהגותיים להפרעת הספקטרום האוטיסטי במודלים של עכברים

Strategies for Assessing Autistic-Like Behaviors in Mice

Autism spectrum disorder (ASD) is a neurobiologically complex condition with a heterogeneous genetic etiology. Clinically, ASD is diagnosed by social communication impairments and restrictive or repetitive behaviors, such as hand flapping or lining up objects. These behavioral patterns can be reliably observed in mouse models with ASD-linked genetic mutations, making them highly useful tools for studying the underlying cellular and molecular mechanisms in ASD. Understanding how genetic changes affect the neurobiology and behaviors observed in ASD will facilitate the development of novel targeted therapeutic compounds to ameliorate core behavioral impairments. Our lab has employed several protocols encompassing well-described training and testing procedures that reflect a wide range of behavioral deficits related to ASD. Here, we detail two assays to study the core features of ASD in mouse models: self-grooming (a measure of repetitive behavior) and the three-chamber social interaction test (a measure of social interaction approach and preference for social novelty).

<meta charset="utf8"></head><body>אסטרטגיות להערכת התנהגויות דמויות אוטיסטיות בעכברים

Rodent models are valuable tools for studying core behaviors related to autism spectrum disorder (ASD). In this article, we expound on two behavioral tests for modeling the core features of ASD in mice: self-grooming, which assesses repetitive behavior, and the three-chamber social interaction test, which documents social impairments.

אסטרטגיות להערכת התנהגויות דמויות אוטיסטיות בעכברים

Autism spectrum disorder (ASD) is a neurobiologically complex condition with a heterogeneous genetic etiology. Clinically, ASD is diagnosed by social ...

Immunohistochemical Visualization of Hippocampal Neuron Activity After Spatial Learning in a Mouse Model of Neurodevelopmental Disorders

Induction of phosphorylated extracellular-regulated kinase (pERK) is a reliable molecular readout of learning-dependent neuronal activation. Here, we describe a pERK immunohistochemistry protocol to study the profile of hippocampal neuron activation following exposure to a spatial learning task in a mouse model characterized by cognitive deficits of neurodevelopmental origin. Specifically, we used pERK immunostaining to study neuronal activation following Morris water maze (MWM, a classical hippocampal-dependent learning task) in Engrailed-2 knockout (En2-/-) mice, a model of autism spectrum disorders (ASD). As compared to wild-type (WT) controls, En2-/- mice showed significant spatial learning deficits in the MWM. After MWM, significant differences in the number of pERK-positive neurons were detected in specific hippocampal subfields of En2-/- mice, as compared to WT animals. Thus, our protocol can robustly detect differences in pERK-positive neurons associated to hippocampal-dependent learning impairment in a mouse model of ASD. More generally, our protocol can be applied to investigate the profile of hippocampal neuron activation in both genetic or pharmacological mouse models characterized by cognitive deficits.

We describe an immunohistochemistry protocol to study the profile of hippocampal neuron activation following exposure to a spatial learning task in a mouse model characterized by cognitive deficits of neurodevelopmental origin. This protocol can be applied to both genetic or pharmacological mouse models characterized by cognitive deficits.

ויזואליזציה immunohistochemical של היפוקמפוס Neuron פעילות לאחר מרחבי למידה במודל עכבר של הפרעות נוירו-התפתחותי

Induction of phosphorylated extracellular-regulated kinase (pERK) is a reliable molecular readout of learning-dependent neuronal activation. Here, we ...

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

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לחקור את המוח באוטיזם באמצעות fMRI ו דימות מותח דיפוזי

בסיס מבוסס על תגובת שרשרת פולימראז חזקה עבור כימות ציטוסינוס-גואנין-גואנין Trinucleotide גאות חוזר בצורה שברירית X פיגור-1 גן

פרוטוקול אלקטרופיזיולוגיה אמצעי תגמול ציפייה, עיבוד אצל ילדים

זיהוי מהיר של פנוטיפים התפתחותיות בתאי האדם קודמן עצבית (NPCs)

חקירות על שינויים בתפקוד המעגל היפוקמפוס בעקבות פגיעה מוחית טראומטית קלה

הקלטות תא כל לזווג בפרוסות בהיפוקמפוס Organotypic

אסטרטגיות להערכת התנהגויות דמויות אוטיסטיות בעכברים

אסטרטגיות להערכת התנהגויות דמויות אוטיסטיות בעכברים

אסטרטגיות להערכת התנהגויות דמויות אוטיסטיות בעכברים

ויזואליזציה immunohistochemical של היפוקמפוס Neuron פעילות לאחר מרחבי למידה במודל עכבר של הפרעות נוירו-התפתחותי