עבודה זו מומנה על ידי איגוד הסוכרת האמריקני (ADA), גרנט JF1-10-14 (MDiF). ברצוננו להודות לכל אנשי המעבדה של בעלי חיים על משאבים WSU. מחבר בתודה להכיר סיוע ניתוח סטטיסטי של נתונים ניל Paton, Ph.D.

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	<li>Baron, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+neuropathic+pain:+From+mechanisms+to+symptoms.">Peripheral neuropathic pain: From mechanisms to symptoms.</a> Clin. J. Pain. 16, S12-S20 (2000).</li><li>Calcutt, N. A., Jorge, M. C., Yaksh, T. L., Chaplan, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tactile+allodynia+and+formalin+hyperalgesia+in+streptozotocin-diabetic+rats:+Effects+of+insulin,+aldose+reductase+inhibition+and+lidocaine.">Tactile allodynia and formalin hyperalgesia in streptozotocin-diabetic rats: Effects of insulin, aldose reductase inhibition and lidocaine.</a> Pain. 68, 293-299 (1996).</li><li>Bolcskei, K., Petho, G., Szolcsanyi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noxious+heat+threshold+measured+with+slowly+increasing+temperatures:+Novel+rat+thermal+hyperalgesia+models.">Noxious heat threshold measured with slowly increasing temperatures: Novel rat thermal hyperalgesia models.</a> Methods Mol. Biol. 617, 57-66 (2010).</li><li>Ankier, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+hot+plate+tests+to+quantify+antinociceptive+and+narcotic+antagonist+activities.">New hot plate tests to quantify antinociceptive and narcotic antagonist activities.</a> Eur. J. Pharmacol. 27, 1-4 (1974).</li><li>Almasi, R., Petho, G., Bolcskei, K., Szolcsanyi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+resiniferatoxin+on+the+noxious+heat+threshold+temperature+in+the+rat:+A+novel+heat+allodynia+model+sensitive+to+analgesics.">Effect of resiniferatoxin on the noxious heat threshold temperature in the rat: A novel heat allodynia model sensitive to analgesics.</a> Br. J. Pharmacol. 139, 49-58 (2003).</li><li>Bolcskei, K., Horvath, D., Szolcsanyi, J., Petho, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heat+injury-induced+drop+of+the+noxious+heat+threshold+measured+with+an+increasing-temperature+water+bath:+A+novel+rat+thermal+hyperalgesia+model.">Heat injury-induced drop of the noxious heat threshold measured with an increasing-temperature water bath: A novel rat thermal hyperalgesia model.</a> Eur. J. Pharmacol. 564, 80-87 (2007).</li><li>Chesler, E. J., Wilson, S. G., Lariviere, W. R., Rodriguez-Zas, S. L., Mogil, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+ranking+of+genetic+and+laboratory+environment+factors+influencing+a+behavioral+trait,+thermal+nociception,+via+computational+analysis+of+a+large+data+archive.">Identification and ranking of genetic and laboratory environment factors influencing a behavioral trait, thermal nociception, via computational analysis of a large data archive.</a> Neurosci. Biobehav. Rev. 26, 907-923 (2002).</li><li>Langford, D. J., Crager, S. E., Shehzad, Z., Smith, S. B., Sotocinal, S. G., Levenstadt, J. S., Chanda, M. L., Levitin, D. J., Mogil, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+modulation+of+pain+as+evidence+for+empathy+in+mice.">Social modulation of pain as evidence for empathy in mice.</a> Science. 312, 1967-1970 (2006).</li><li>Hunt, S. P., Koltzenburg, M. <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 neurobiology of pain. , (2005).</li><li>Willis, W. D. <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 pain system : The neural basis of nociceptive transmission in the mammalian nervous system. , (1985).</li><li>Calcutt, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Modeling diabetic sensory neuropathy in rats. In: Methods in molecular medicine. Pain research: Methods and protocols. , (2004).</li><li>Bars, D. L. e., Gozariu, M., Cadden, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+nociception.">Animal models of nociception.</a> Pharmacol. Rev. 53, 597-652 (2001).</li><li>Shaikh, A. S., Somani, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+and+biomarkers+of+neuropathy+in+diabetic+rodents.">Animal models and biomarkers of neuropathy in diabetic rodents.</a> Indian J. Pharmacol. 42, 129-134 (2010).</li><li>Hargreaves, K., Dubner, R., Brown, F., Flores, C., Joris, 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+new+and+sensitive+method+for+measuring+thermal+nociception+in+cutaneous+hyperalgesia.">A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia.</a> Pain. 32, 77-88 (1988).</li><li>Hardy, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+for+the+rapid+measurement+of+skin+temperature+during+exposure+to+intense+thermal+radiation.">Method for the rapid measurement of skin temperature during exposure to intense thermal radiation.</a> J. Appl. Physiol. 5, 559-566 (1953).</li><li>Sumino, R., Dubner, R., Starkman, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+small+myelinated+&amp;quot;warm&amp;quot;+fibers+to+noxious+heat+stimuli+applied+to+the+monkey's+face.">Responses of small myelinated &amp;quot;warm&amp;quot; fibers to noxious heat stimuli applied to the monkey's face.</a> Brain Res. 62, 260-263 (1973).</li><li>Hammond, D. L., Ruda, 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=Developmental+alterations+in+thermal+nociceptive+threshold+and+the+distribution+of+immunoreactive+calcitonin+gene-related+peptide+and+substance+p+after+neonatal+administration+of+capsaicin+in+the+rat.">Developmental alterations in thermal nociceptive threshold and the distribution of immunoreactive calcitonin gene-related peptide and substance p after neonatal administration of capsaicin in the rat.</a> Neurosci. Lett. 97, 57-62 (1989).</li></ol>

החלטנו לא לחשוף את זהותו של כאבים השתמשו בניסויים אלו. חלק מן התוצאות המוצגות כאן הוצגה ב -71 מושבים רחוב המדעיים של איגוד הסוכרת האמריקני (סן דייגו קליפורניה) ואת הכנס השנתי ה 47 של האגודה האירופית לחקר הסוכרת (ליסבון, פורטוגל).

באופן דומה למבחן הצלחת קלאסי חם לכמת hyperalgesia תרמית 4,13, assay של nociception המתואר כאן מאפשר דרך מהירה ואמינה לכמת התנהגות nocifensive בחולדות ועכברים. עם זאת, בניגוד מבחן קלאסי, השיטה המצטברת צלחת חמה אינו פולשני ולמעשה ללא מתח. למרות שחלק לרסן יש צורך לבצע את הבדיקה (כלומר,...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> שם </td><td> חברה </td><td> מספר קטלוגי </td></tr><tr><td> מצטבר חם פלייט שיכוך כאבים Meter </td><td> IITC מדעי החיים בע&quot;מ </td><td> חלק # PE34 </td></tr><tr><td> סדרת Soft 8 </td><td> IITC מדעי החיים בע&quot;מ </td><td> חלק # Series8 </td></tr><tr><td> Streptozotocin </td><td> Calbiochem </td><td> 572201 </td></tr></tbody></table>

rapid determination of the thermal nociceptive threshold in diabetic rats

נוירופתיה סוכרתית כואבת (PDN) מאופיין hyperalgesia כלומר, רגישות מוגברת לגירוי מזיק, ו allodynia כלומר, רגישות יתר לגירויים לא מזיקים בדרך כלל 1. Hyperalgesia ו allodynia נחקרו רבות מכרסמים מודלים שונים של סוכרת 2. עם זאת, כאמור על ידי Bölcskei ואח&#39;, קביעת &quot;כאב&quot; במודלים של בעלי חיים הוא מאתגר בשל אופי סובייקטיבי שלה 3. יתר על כן, השיטות המסורתיות ששימשו בקביעת תגובות התנהגותיות לגירויים תרמית רעילים בדרך כלל אין שחזור ו 3 רגישות תרופתי. למשל, באמצעות השיטה צלחת חמה של Ankier 4, נרתע, ונסיגה / או ליקוק של או אחורית ו / או האסיפות כפות מכמתים כמו latencies רפלקס ב גירויים קבועים תרמית גבוהה (52-55 מעלות צלזיוס). עם זאת, בעלי חיים, כי הם hyperalgesic לגירוי תרמי לא reproducibly להראות הבדלים מחדשלהגמיש latencies באמצעות אלה לעיל סף בטמפרטורות 3,5. כמו השיטה המתוארת לאחרונה Bölcskei et al. 6, בהתאם לנוהל המתואר מאפשר קביעה מהירה, רגיש לשחזור של ספי nociceptive תרמית (TNTs) בעכברים וחולדות. השיטה משתמשת גירוי תרמית לאט להגדיל מיושם בעיקר על העור של משטח העכבר / עכברוש plantar. השיטה היא רגישה במיוחד ללמוד אנטי nociception במהלך מדינות hyperalgesic כגון PDN. הנהלים שתוארו לשאוג מבוססים על אלה שפורסמו בהרחבה על ידי Almási et al 5 ו Bölcskei ואח&#39; 3. הנהלים שתוארו כאן אושרו על בעלי חיים במעבדה טיפול ועדת שימוש (LACUC), רייט סטייט.

Watch this Scientific Journal Video about Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats at JoVE.com

Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats

כאן אנו מתארים הליך אמין ופשוט מהירה כדי לקבוע את הטמפרטורה הנמוכה ביותר שבה חולדות או עכברים מראים התנהגות nocifensive,<em&gt; כלומר</em&gt;<em&gt; סף nociceptive תרמית</em&gt; (TNT). שיטה זו חל גירוי תרמית לאט להגדיל המאפשר הערכה מדויקת לשחזור של TNTs עם מינימום, אם בכלל, הלחץ על בעלי חיים.

קביעה מהירה של סף nociceptive תרמי בעכברים סוכרתיים

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity ...

rapid-determination-thermal-nociceptive-threshold-diabetic

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Neuroscience

27.4K Views.  Wright State University. כאן אנו מתארים הליך אמין ופשוט מהירה כדי לקבוע את הטמפרטורה הנמוכה ביותר שבה חולדות או עכברים מראים התנהגות nocifensive, כלומר סף nociceptive תרמית (TNT). שיטה זו חל גירוי תרמית לאט להגדיל המאפשר הערכה מדויקת לשחזור של TNTs עם מינימום, אם בכלל, הלחץ על בעלי חיים.

Video: Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

שיטה כללית להערכת דגירה של השתוקקות סוכרוז בחולדות

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

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 ...

Assessment of Morphine-induced Hyperalgesia and Analgesic Tolerance in Mice Using Thermal and Mechanical Nociceptive Modalities

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular mechanisms underlying both phenomena are not well understood and their elucidation should benefit from the study of animal models and from the design of appropriate experimental protocols. 
We describe here a methodological approach for inducing, recording and quantifying morphine-induced hyperalgesia as well as for evidencing analgesic tolerance, using the tail-immersion and tail pressure tests in wild-type mice. As shown in the video, the protocol is divided into five sequential steps. Handling and habituation phases allow a safe determination of the basal nociceptive response of the animals. Chronic morphine administration induces significant hyperalgesia as shown by an increase in both thermal and mechanical sensitivity, whereas the comparison of analgesia time-courses after acute or repeated morphine treatment clearly indicates the development of tolerance manifested by a decline in analgesic response amplitude. This protocol may be similarly adapted to genetically modified mice in order to evaluate the role of individual genes in the modulation of nociception and morphine analgesia. It also provides a model system to investigate the effectiveness of potential therapeutic agents to improve opiate analgesic efficacy.

We describe a protocol to examine the development of opioid-induced hyperalgesia and tolerance in mice. Based on the measurement of thermal and mechanical nociceptive responses of na&#239;ve and morphine-treated animals, it allows to quantify the increase in pain sensitivity (hyperalgesia) and decrease in analgesia (tolerance) associated with chronic opiate administration.

הערכה של סובלנות hyperalgesia ומשככי כאבים הנגרם מורפיום בעכברים באמצעות שיטות nociceptive תרמיים ומכאניות

Opioid-induced hyperalgesia and tolerance severely impact the clinical efficacy of opiates as pain relievers in animals and humans. The molecular ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such behavior is spontaneous locomotor activity. Here we describe our protocol that utilizes Drosophila population monitors and a tracking system that allows continuous monitoring of the spontaneous locomotor activity of flies for several days at a time. This method is simple, reliable, and objective and can be used to examine the effects of aging, sex, changes in caloric content of food, addition of drugs, or genetic manipulations that mimic human diseases.

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.&#160; Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.&#160;

קביעת הפעילות של תנועה הספונטנית ב<em&gt; Melanogaster דרוזופילה</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome

3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) toxicity may cause region-specific changes in serotonergic mRNA expression due to acute serotonin (5-hydroxytryptamine; 5-HT) syndrome. This hypothesis can be tested using in situ hybridization to detect the serotonin 5-HT2A receptor gene htr2a. In the past, such procedures, utilizing radioactive riboprobe, were difficult because of the complicated workflow that needs several days to perform and the added difficulty that the technique required the use of fresh frozen tissues maintained in an RNase-free environment. Recently, the development of short oligonucleotide probes has simplified in situ hybridization procedures and allowed the use of paraformaldehyde-prefixed brain sections, which are more widely available in laboratories. Here, we describe a detailed protocol using non-radioactive oligonucleotide probes on the prefixed brain tissues. Hybridization probes used for this study include dapB (a bacterial gene coding for dihydrodipicolinate reductase), ppiB (a housekeeping gene coding for peptidylprolyl isomerase B), and htr2a (a serotonin gene coding for 5-HT2A receptors). This method is relatively simply, cheap, reproducible and requires less than two days to complete.

Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome

This protocol describes a rapid and simplified in situ hybridization method ideal forparaformaldehyde-prefixed brain, thus reducing the need for prolonged complex steps while using fresh frozen tissues. The method is validated using the identification of the serotonin 5-HT2A receptor gene htr2a in rats.

מהיר<em&gt; באתר</em&gt; הכלאה באמצעות oligonucleotide בדיקות במוח-התחילית Paraformaldehyde של חולדות עם סרוטונין תסמונת

3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) toxicity may cause region-specific changes in serotonergic mRNA expression due to acute serotonin ...

Measurement &amp; Analysis of the Temporal Discrimination Threshold Applied to Cervical Dystonia

The temporal discrimination threshold (TDT) is the shortest time interval at which an observer can discriminate two sequential stimuli as being asynchronous (typically 30-50 ms). It has been shown to be abnormal (prolonged) in neurological disorders, including cervical dystonia, a phenotype of adult onset idiopathic isolated focal dystonia. The TDT is a quantitative measure of the ability to perceive rapid changes in the environment and is considered indicative of the behavior of the visual neurons in the superior colliculus, a key node in covert attentional orienting. This article sets out methods for measuring the TDT (including two hardware options and two modes of stimuli presentation). We also explore two approaches of data analysis and TDT calculation. The application of the assessment of temporal discrimination to the understanding of the pathogenesis of cervical dystonia and adult onset idiopathic isolated focal dystonia is also discussed.

Methods for the measurement and analysis of the temporal discrimination threshold are presented, and its application to the study of the pathogenesis of cervical dystonia are discussed.

מדידה &amp; ניתוח של הסף אפליה זמני המוחל פוקאלית צוואר הרחם

The temporal discrimination threshold (TDT) is the shortest time interval at which an observer can discriminate two sequential stimuli as being ...

Rapid and Specific Immunomagnetic Isolation of Mouse Primary Oligodendrocytes

The efficient and robust isolation and culture of primary oligodendrocytes (OLs) is a valuable tool for the in vitro study of the development of oligodendroglia as well as the biology of demyelinating diseases such as multiple sclerosis and Pelizaeus-Merzbacher-like disease (PMLD). Here, we present a simple and efficient selection method for the immunomagnetic isolation of stage three O4+ preoligodendrocytes cells from neonatal mice pups. Since immature OL constitute more than 80% of the rodent-brain white matter at postnatal day 7 (P7) this isolation method not only ensures high cellular yield, but also the specific isolation of OLs already committed to the oligodendroglial lineage, decreasing the possibility of isolating contaminating cells such as astrocytes and other cells from the mouse brain. This method is a modification of the techniques reported previously, and provides oligodendrocyte preparation purity above 80% in about 4 h.

We describe the immunomagnetic isolation of primary mouse oligodendrocytes, which allows the rapid and specific isolation of the cells for in vitro culture.

בידוד Immunomagnetic מהירה, מדויקת של העכבר הראשי oligodendrocytes להפוך

The efficient and robust isolation and culture of primary oligodendrocytes (OLs) is a valuable tool for the in vitro study of the development of ...

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided here is a protocol for successfully establishing a rat model of acute pontine infarction. Rats weighing about 250 g are used, and a probe with an insulated sheath is injected into the pons using a stereotaxic apparatus. A lesion is produced by the electrical stimulation with a single pulse. The Longa score, Berderson score, and beam balance test are used to assess neurological deficits. Additionally, the adhesive-removal somatosensory test is used to determine sensorimotor function, and the limb placement test is used to evaluate proprioception. MRI scans are then used to assess the infarction in vivo, and TTC staining is used to confirm the infarction in vitro. Here, a successful infarction is identified that is located in the anterolateral basis of the rostral pons. In conclusion, a new method is described to establish an acute pontine infarction rat model.

Presented here is a protocol for establishing acute pontine infarction in a rat model via electrical stimulation with a single pulse.

הקמת אוטם פונטינה חריפה בחולדות על ידי גירוי חשמלי

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided ...

Laser-Induced Brain Injury in the Motor Cortex of Rats

A common technique for inducing stroke in experimental rodent models involves the transient (often denoted as MCAO-t) or permanent (designated as MCAO-p) occlusion of the middle cerebral artery (MCA) using a catheter. This generally accepted technique, however, has some limitations, thereby limiting its extensive use. Stroke induction by this method is often characterized by high variability in the localization and size of the ischemic area, periodical occurrences of hemorrhage, and high death rates. Also, the successful completion of any of the transient or permanent procedures requires expertise and often lasts for about 30 minutes. In this protocol, a laser irradiation technique is presented that can serve as an alternative method for inducing and studying brain injury in rodent models.
When compared to rats in the control and MCAO groups, the brain injury by laser induction showed reduced variability in body temperature, infarct volume, brain edema, intracranial hemorrhage, and mortality. Furthermore, the use of a laser-induced injury caused damage to the brain tissues only in the motor cortex unlike in the MCAO experiments where destruction of both the motor cortex and striatal tissues is observed.
Findings from this investigation suggest that laser irradiation could serve as an alternative and effective technique for inducing brain injury in the motor cortex. The method also shortens the time for completing the procedure and does not require expert handlers.

The protocol presented here shows a technique to create a rodent model of brain injury. The method described here uses laser irradiation and targets motor cortex.

פגיעה מוחית כתוצאה בלייזר בקליפת המוח המוטורית של חולדות

A common technique for inducing stroke in experimental rodent models involves the transient (often denoted as MCAO-t) or permanent (designated as MCAO-p) ...

Induction of Diffuse Axonal Brain Injury in Rats Based on Rotational Acceleration

Traumatic brain injury (TBI) is a major cause of death and disability. Diffuse axonal injury (DAI) is the predominant mechanism of injury in a large percentage of TBI patients requiring hospitalization. DAI involves widespread axonal damage from shaking, rotation or blast injury, leading to rapid axonal stretch injury and secondary axonal changes that are associated with a long-lasting impact on functional recovery. Historically, experimental models of DAI without focal injury have been difficult to design. Here we validate a simple, reproducible and reliable rodent model of DAI that causes widespread white matter damage without skull fractures or contusions.

This protocol validates a reliable, easy-to-perform and reproducible rodent model of brain diffuse axonal injury (DAI) that induces widespread white matter damage without skull fractures or contusions.

אינדוקציה של פגיעה מוחית Axonal בחולדות מבוסס על האצת הסיבוב

Traumatic brain injury (TBI) is a major cause of death and disability. Diffuse axonal injury (DAI) is the predominant mechanism of injury in a large ...