נתמך על ידי אוניברסיטת קנטקי, החוג לביולוגיה, משרד לימודי הסמכה ו במכללה לאמנויות ומדעים.

<ol>
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אין ניגודי אינטרסים הכריז.

הפרטים הניתנים בסרט הקשורים טקסט סיפקו שלבים עיקריים על מנת מספיק להקליט את הפעילות MRO של סרטנים באתרם. אחת ממטרות הדו&quot;ח שלנו היא להגדיל את המודעות לפוטנציאל של הכנה זו התלמיד להפעיל מעבדות חקירה ללמד מושגי יסוד בפיזיולוגיה של החושים. ההכנות הם חזקים מאוד כדאיות בעת היותו שטוף מלוחים מינימלי. שליטה מוטורית על שרירי MRO זוהתה אך תקנה בשידור ואת הפוטנציאל של הפלסטיות הסינפטית כמו גם היעילות של שידור עבור נוירונים מעוררים ו מעכבות נותר שטח פתוח לחקירה (Elekes ו Florey, 1987a, b; Florey ו Florey, 1955; Kuffler, 1954; Kuffler ו Eyzaguirre, 1955). הכנה זו יכולה לשמש כדי לחקור מספר תנאי הניסוי, כמו גם את מגוון הטבעי של תנועה בתוך החיה כדי להבין טוב יותר את הביולוגיה של MRO למחקר ראשוני, כמו גם למטרות הפגנתי. המאפיינים הללו biophysical עצב סנסורי יש בחלק טופלו בטבע שלהם הסתגלות בפעילות העצבית עם גירוי שמר (Brown et al, 1978; Edwards et al, 1981;. Purali, 1997; Rydqvist ו Purali, 1991; Rydqvist ו Swerup, 1991). עם זאת, רק מעטים כתובת neuromodulation דיווחים על קולטנים אלה חושי סיבי שריר הקשורים (Cooper et al, 2003;. פסטור ו מקמילן, 1990). הדוחות להתמודד עם רק כמה תרכובות רבות שידוע כי הם להיות נוכחים hemolymph. מאפננים רבים של קוקטיילים מאפננים להישאר להיבדק על מורכבות MRO (השרירים נוירונים). פסטור ו מקמילן (1990) היה לבחון את neuromodulators 5-HT ו octopamine על פעילות MROs בין מינים שונים סרטנאים וציין כי ישנם הבדלים מינים. הם לא לבחון בפירוט את ההשפעות לטווח ארוך של neuromodulators אלה ולא את ההשפעה על הפעילות בעמדות שונות של MRO. זה סוג של הכנה יכול לסייע בהבנת הבסיס של תפיסה ויסות חושי של עיבוד עצביים אשר חשוב בשיקום וניהול המחלה לבני אדם עם abnormities יחידת המנוע (פאטל et al, 2009;. רבין et al, 2009;. מרינו ואח&#39; . 2010). סוגים שונים של קלט דפוסי ירי לצורך ניטור תנועות המפרק בהכנות חוליות נגיש ניתן להשתמש רובוטיקה / תותבת (מקמילן Patullo, 2001). יש עדיין שאלות רבות ממתינה לתשובות כהכנה זה יכול להועיל במספר דרכים.

<ol>
<li><a href="http://www.adinstruments.com/products/hardware/corporate/product/ML856/" target="_blank">PowerLab 26T (AD Instruments)</a> </li>
<li><a href="http://www.adinstruments.com/products/software/research/LabChart-Software/" target="_blank">LabChart 7 (ADI Instruments, Colorado Springs, CO, USA)</a> </li>
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muscle receptor organs in the crayfish abdomen: a student laboratory exercise in proprioception

המטרה העיקרית של הניסוי היא להוכיח העיקרי להעברת מידע עצב סנסורי של תנועות ותנוחות המשותפת מידע פרופריוצפטיביות עבור חיה. מטרה נוספת של הניסוי הזה היא ללמוד אנטומיה של התכשיר על ידי לנתיחה מכתים ועל ההצגה של נוירונים ומבנים חושי תחת מיקרוסקופ לנתח. זו מתבצעת באמצעות ציוד בסיסי neurophysiological כדי להקליט את הפעילות החשמלית מאיבר קולטן משותף וטכניקות צביעה. קולטן שריר איבר (MRO) מערכת סרטנים משול כישור השריר intrafusal ביונקים, אשר מסייע לשמש מודל השוואתי כי הוא יותר נגיש עבור הקלטות אלקטרו. בנוסף, אלה הם עצב סנסורי לזיהוי בין ההכנות. ההכנה היא קיימא מלוחים מינימלית במשך שעות וזה מקובל על תרגילי מעבדה סטודנט. MRO היא גם רגישים neuromodulation המעודדת שאלות מסקרנות באתרים פעולה modulatory ושילוב דינמי של אותות של תנועות ומיקום סטטי יחד עם רווח כי ניתן לשנות את המערכת.

Watch this Scientific Journal Video about Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception at JoVE.com

Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception

המטרה העיקרית של הניסוי היא להבין כיצד עצב סנסורי עיקרי להעברת מידע על תנועות ותנוחות המשותפת מידע פרופריוצפטיביות עבור חיה. מטרה נוספת של דו&quot;ח זה היא להציג את האנטומיה של התכשיר על ידי לנתיחה וצפייה של נוירונים תחת מיקרוסקופ לנתח.

קולטן שרירים איברים בבטן סרטן הנהרות: מעבדה הסטודנטים תרגיל הפרופריוספציה

The primary purpose of this experiment is to demonstrate primary sensory neurons conveying information of joint movements and positions as proprioceptive ...

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22.1K Views.  University of Kentucky. המטרה העיקרית של הניסוי היא להבין כיצד עצב סנסורי עיקרי להעברת מידע על תנועות ותנוחות המשותפת מידע פרופריוצפטיביות עבור חיה. מטרה נוספת של דו"ח זה היא להציג את האנטומיה של התכשיר על ידי לנתיחה וצפייה של נוירונים תחת מיקרוסקופ לנתח.

Video: Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception

Recording Behavioral Responses to Reflection in Crayfish

Social behavior depends on sensory input from the visual, mechanical and olfactory systems. One important issue concerns the relative roles of each sensory modality in guiding behavior. The role of visual inputs has been examined by isolating visual stimuli from mechanical and chemosensory stimuli. In some studies (Bruski &amp; Dunham, 1987: Delgado-Morales et al., 2004) visual inputs have been removed with blindfolds or low light intensity, and effects of remaining sensory modalities have been elucidated. An alternative approach is to study the effects of visual inputs in the absence of any appropriate mechanical and chemosensory cues. This approach aims to identify the exclusive role of visual inputs. 
We have used two methods to provide visual stimuli to crayfish without providing chemical and mechanical cues. In one method, crayfish are videotaped in an aquarium where half of the walls are covered in mirrors to provide a reflective environment, and the other half are covered in a non-reflective (matte finish) plastic. This gives the crayfish a choice between reflective and non-reflective environments. The reflective environment provides visual cues in the form of reflected images of the crayfish as it moves throughout half of the tank; these visual cues are missing from the non-reflective half of the tank. An alternative method is to videotape the behavior of crayfish in an aquarium separated by a smaller chamber at each end, with a crayfish in one small chamber providing visual cues and an inert object in the opposite small chamber providing visual input from a non-moving, non-crayfish source. 
Our published results indicate that responses of crayfish to the reflective environment depend on socialization and dominance rank. Socialized crayfish spent more time in the reflective environment and exhibited certain behaviors more frequently there than in the non-reflective environment; isolated crayfish showed no such differences. Crayfish that were housed in same-sex pairs developed a social rank of either dominant or subordinate. Responses to reflection differed between dominant and subordinate crayfish (May &amp; Mercier, 2006; May &amp; Mercier, 2007). Dominant crayfish spent more time on the reflective side, entered reflective corners more frequently and spent more time in reflective corners compared to the non-reflective side. Subordinate crayfish walked in reverse more often on the reflective side than on the non-reflective side. Preliminary data suggest similar effects from visual cues provided by a crayfish in a small adjoining chamber (May et al., 2008).

We have developed two methods for studying effects of visual cues on behavior in the absence of tactile and chemical cues. One method involves videotaping responses of crayfish to reflective walls in an aquarium; the other examines effects of visual inputs provided by a live crayfish behind a transparent partition.

הקלטה תגובות התנהגותיות כדי השתקפות של סרטן הנהרות

Social behavior depends on sensory input from the visual, mechanical and olfactory systems.  One important issue concerns the relative roles of each ...

A Procedure for Studying the Footshock-Induced Reinstatement of Cocaine Seeking in Laboratory Rats

The most insidious aspect of drug addiction is the high propensity for relapse. Animal models of relapse, known as reinstatement procedures, have been used extensively to study the neurobiology and phenomenology of relapse to drug use. Although procedural variations have emerged over the past several decades, the most conventional reinstatement procedures are based on the drug self-administration (SA) model. In this model, an animal is trained to perform an operant response to obtain drug. Subsequently, the behavior is extinguished by withholding response-contingent reinforcement. Reinstatement of drug seeking is then triggered by a discrete event, such as an injection of the training drug, re-exposure to drug-associated cues, or exposure to a stressor 1.
Reinstatement procedures were originally developed to study the ability of acute non-contingent exposure to the training drug to reinstate drug seeking in rats and monkeys 1, 2. Reinstatement procedures have since been modified to study the role of environmental stimuli, including drug-associated cues and exposure to various forms of stress, in relapse to drug seeking 1, 3, 4.
Over the past 15 years, a major focus of the reinstatement literature has been on the role of stress in drug relapse. One of the most commonly used forms of stress for studying this relationship is acute exposures to mild, intermittent, electric footshocks. The ability of footshock stress to induce reinstatement of drug seeking was originally demonstrated by Shaham and colleagues (1995) in rats with a history of intravenous heroin SA5. Subsequently, the effect was generalized to rats with histories of intravenous cocaine, methamphetamine, and nicotine SA, as well as oral ethanol SA 3, 6.
Although footshock-induced reinstatement of drug seeking can be achieved reliably and robustly, it is an effect that tends to be sensitive to certain parametrical variables. These include the arrangement of extinction and reinstatement test sessions, the intensity and duration of footshock stress, and the presence of drug-associated cues during extinction and testing for reinstatement. Here we present a protocol for footshock-induced reinstatement of cocaine seeking that we have used with consistent success to study the relationship between stress and cocaine seeking.

Animal models of relapse, known as reinstatement procedures, have been used extensively to study the role of stress in relapse to drug seeking. Here, we report on a method for inducing the reinstatement of cocaine seeking in laboratory rats via acute exposures to mild, intermittent electric footshock.

נוהל לימוד החזרה Footshock-induced של קוקאין מחפשים אצל חולדות מעבדה

The most insidious aspect of drug addiction is the high propensity for relapse.  Animal models of relapse, known as reinstatement procedures, have been ...

Physiological Recordings of High and Low Output NMJs on the Crayfish Leg Extensor Muscle

We explain in detail how to expose and conduct electrophysiological recordings of synaptic responses for high (phasic) and low (tonic) output motor neurons innervating the extensor muscle in the walking leg of a crayfish. Distinct differences are present in the physiology and morphology of the phasic and tonic nerve terminals. The tonic axon contains many more mitochondria, enabling it to take a vital stain more intensely than the phasic axon. The tonic terminals have varicosities, and the phasic terminal is filiform. The tonic terminals are low in synaptic efficacy but show dramatic facilitated responses. In contrast, the phasic terminals are high in quantal efficacy but show synaptic depression with high frequency stimulation. The quantal output is measured with a focal macropatch electrode placed directly over the visualized nerve terminals. Both phasic and tonic terminals innervate the same muscle fibers, which suggests that inherent differences in the neurons, rather than differential retrograde feedback from the muscle, account for the morphological and physiological differentiation.

This article demonstrates how to conduct electrophysiological recordings of synaptic responses on the extensor muscle in the walking leg of a crayfish and how the nerve terminals are visualized to show the gross morphological differences of high- and low-output nerve terminals.

הקלטות פיזיולוגיים של NMJs פלט גבוה ונמוך על שרירים ברגליים סרטן הנהרות extensor

We explain in detail how to expose and conduct electrophysiological recordings of synaptic responses for high (phasic) and low (tonic) output motor ...

Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises

The purpose of this report is to help develop an understanding of the effects caused by ion gradients across a biological membrane. Two aspects that influence a cell's membrane potential and which we address in these experiments are: (1) Ion concentration of K+ on the outside of the membrane, and (2) the permeability of the membrane to specific ions. The crayfish abdominal extensor muscles are in groupings with some being tonic (slow) and others phasic (fast) in their biochemical and physiological phenotypes, as well as in their structure; the motor neurons that innervate these muscles are correspondingly different in functional characteristics. We use these muscles as well as the superficial, tonic abdominal flexor muscle to demonstrate properties in synaptic transmission. In addition, we introduce a sensory-CNS-motor neuron-muscle circuit to demonstrate the effect of cuticular sensory stimulation as well as the influence of neuromodulators on certain aspects of the circuit. With the techniques obtained in this exercise, one can begin to answer many questions remaining in other experimental preparations as well as in physiological applications related to medicine and health. We have demonstrated the usefulness of model invertebrate preparations to address fundamental questions pertinent to all animals.

The experiments demonstrate an easy approach for students to gain experience in examining muscle structure, synaptic responses, the effects of ion gradients and permeability on membrane potentials. Also, a sensory-CNS-motor-muscle circuit is presented to show a means to test effects of compounds on a neuronal circuit.

ממברנה פוטנציאלים, התגובות Synaptic, המעגלים הנוירונים, Neuromodulation ו היסטולוגיה שרירים שימוש סרטן הנהרות: מעבדה תרגילים סטודנטים

The purpose of this report is to help develop an understanding of the effects caused by ion gradients across a biological membrane. Two aspects that ...

Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise

The purpose of the report is to describe dissection techniques for preparing the crayfish hindgut and to demonstrate how to make physiological recordings with a force transducer to monitor the strength of contraction. In addition, we demonstrate how to visually monitor peristaltic activity, which can be used as a bioassay for various peptides, biogenic amines and neurotransmitters. This preparation is amenable to student laboratories in physiology and for demonstrating pharmacological concepts to students. This preparation has been in use for over 100 years, and it still offers much as a model for investigating the generation and regulation of peristaltic rhythms and for describing the mechanisms underlying their modulation. The pharmacological assays and receptor sub-typing that were started over 50 years ago on the hindgut still contribute to research today. This robust preparation is well suited to training students in physiology and pharmacology.

In this report we demonstrate techniques that can be used to investigate the biology of the crayfish hindgut. We show how to dissect a crayfish abdomen and study the associated anatomy, physiology and modulation of activity. The peristaltic activity and strength of contractions are measured using a force transducer.

ניסויים פיזיולוגיים עם במעי האחורי סרטן הנהרות: תרגיל מעבדה סטודנטים

The purpose of the report is to describe dissection techniques for preparing the crayfish hindgut and to demonstrate how to make physiological recordings ...

Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons

A better understanding of the mechanisms governing receptor trafficking between the plasma membrane (PM) and intracellular compartments requires an experimental approach with excellent spatial and temporal resolutions. Moreover, such an approach must also have the ability to distinguish receptors localized on the PM from those in intracellular compartments. Most importantly, detecting receptors in a single vesicle requires outstanding detection sensitivity, since each vesicle carries only a small number of receptors. Standard approaches for examining receptor trafficking include surface biotinylation followed by biochemical detection, which lacks both the necessary spatial and temporal resolutions; and fluorescence microscopy examination of immunolabeled surface receptors, which requires chemical fixation of cells and therefore lacks sufficient temporal resolution1-6 . To overcome these limitations, we and others have developed and employed a new strategy that enables visualization of the dynamic insertion of receptors into the PM with excellent spatial and temporal resolutions 7-17 . The approach includes tagging of a pH-sensitive GFP, the superecliptic pHluorin 18, to the N-terminal extracellular domain of the receptors. Superecliptic pHluorin has the unique property of being fluorescent at neutral pH and non-fluorescent at acidic pH (pH &lt; 6.0). Therefore, the tagged receptors are non-fluorescent when within the acidic lumen of intracellular trafficking vesicles or endosomal compartments, and they become readily visualized only when exposed to the extracellular neutral pH environment, on the outer surface of the PM. Our strategy consequently allows us to distinguish PM surface receptors from those within intracellular trafficking vesicles. To attain sufficient spatial and temporal resolutions, as well as the sensitivity required to study dynamic trafficking of receptors, we employed total internal reflection fluorescent microscopy (TIRFM), which enabled us to achieve the optimal spatial resolution of optical imaging (~170 nm), the temporal resolution of video-rate microscopy (30 frames/sec), and the sensitivity to detect fluorescence of a single GFP molecule. By imaging pHluorin-tagged receptors under TIRFM, we were able to directly visualize individual receptor insertion events into the PM in cultured neurons. This imaging approach can potentially be applied to any membrane protein with an extracellular domain that could be labeled with superecliptic pHluorin, and will allow dissection of the key detailed mechanisms governing insertion of different membrane proteins (receptors, ion channels, transporters, etc.) to the PM.

By tagging the extracellular domains of membrane receptors with superecliptic pHluorin, and by imaging these fusion receptors in cultured mouse neurons, we can directly visualize individual vesicular insertion events of the receptors to the plasma membrane. This technique will be instrumental in elucidating the molecular mechanisms governing receptor insertion to the plasma membrane.

הדמית pHluorin-תויגה הכנסת קולט לממברנה הפלזמה בנוירונים עכבר מתורבתים ראשיים

A better understanding of the mechanisms governing receptor trafficking between the plasma membrane (PM) and intracellular compartments requires an ...

Proprioception and Tension Receptors in Crab Limbs: Student Laboratory Exercises

The primary purpose of these procedures is to demonstrate for teaching and research purposes how to record the activity of living primary sensory neurons responsible for proprioception as they are detecting joint position and movement, and muscle tension. Electrical activity from crustacean proprioceptors and tension receptors is recorded by basic neurophysiological instrumentation, and a transducer is used to simultaneously measure force that is generated by stimulating a motor nerve. In addition, we demonstrate how to stain the neurons for a quick assessment of their anatomical arrangement or for permanent fixation. Staining reveals anatomical organization that is representative of chordotonal organs in most crustaceans. Comparing the tension nerve responses to the proprioceptive responses is an effective teaching tool in determining how these sensory neurons are defined functionally and how the anatomy is correlated to the function. Three staining techniques are presented allowing researchers and instructors to choose a method that is ideal for their laboratory.

Physiological and anatomical techniques are demonstrated to address function and structure for joint proprioceptors and muscle tension receptors in crustacean walking limbs.

רצפטורים פרופריוספציה והמתח בסרטן איברים: תרגילי מעבדה הסטודנטים

The primary purpose of these procedures is to demonstrate for teaching and research purposes how to record the activity of living primary sensory neurons ...

Measuring Oral Fatty Acid Thresholds, Fat Perception, Fatty Food Liking, and Papillae Density in Humans

Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty acid receptors housed on taste cells. Previous research has shown that an individual&#39;s oral sensitivity to fatty acid, specifically oleic acid (C18:1) is associated with body mass index (BMI), dietary fat consumption, and the ability to identify fat in foods. We have developed a reliable and reproducible method to assess oral chemoreception of fatty acids, using a milk and C18:1 emulsion, together with an ascending forced choice triangle procedure. In parallel, a food matrix has been developed to assess an individual&#39;s ability to perceive fat, in addition to a simple method to assess fatty food liking. As an added measure tongue photography is used to assess papillae density, with higher density often being associated with increased taste sensitivity.

Oral chemoreception of fatty acids and the association with diet and fatty food preferences may enable the identification of mechanisms involved with the development of obesity and why dietary changes may be difficult for many individuals.

מדידה אוראלי חומצת שומן ספים, תפיסת שומן, שומן מזון טעם, וצפיפות אדומות בבני אדם

Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this information to the brain. As such, determining how olfactory information is transferred to the brain, modifying both perception and behavior, merits investigation. Interestingly, there is emerging evidence that cellular transduction and transcriptional factors are involved in the diversification of olfactory receptor neuron. Here we provide a robust whole mount immunological labeling method to assay in vivo olfactory receptor neuron organization. Using this method, we identified all olfactory receptor neurons with anti-ELAV antibody, a known pan-neural marker and Or49a-mCD8::GFP, an olfactory receptor neuron specifically expressed in Nba neuron using anti-GFP antibody.

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Herein we describe the process of whole mount immunostaining of Drosophila antennae, which enables us to better understand the molecular mechanisms involved in the diversification of olfactory receptor neurons (ORN)s.

הר immunolabeling השלם של חוש הריח קולטן נוירונים<em&gt; דרוזופילה</em&gt; אנטנה

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

Adaptation of Microelectrode Array Technology for the Study of Anesthesia-induced Neurotoxicity in the Intact Piglet Brain

Every year, millions of children undergo anesthesia for a multitude of procedures. However, studies in both animals and humans have called into question the safety of anesthesia in children, implicating anesthetics as potentially toxic to the brain in development. To date, no studies have successfully elucidated the mechanism(s) by which anesthesia may be neurotoxic. Animal studies allow investigation of such mechanisms, and neonatal piglets represent an excellent model to study these effects due to their striking developmental similarities to the human brain.
This protocol adapts the use of enzyme-based microelectrode array (MEA) technology as a novel way to study the mechanism(s) of anesthesia-induced neurotoxicity (AIN). MEAs enable real-time monitoring of in vivo neurotransmitter activity and offer exceptional temporal and spatial resolution. It is hypothesized that anesthetic neurotoxicity is caused in part by glutamate dysregulation and MEAs offer a method to measure glutamate. The novel implementation of MEA technology in a piglet model presents a unique opportunity for the study of AIN.

This study explores the novel use of enzyme-based microelectrode array (MEA) technology to monitor in vivo neurotransmitter activity in piglets. The hypothesis was that glutamate dysregulation contributes to the mechanism of anesthetic neurotoxicity. Here, we present a protocol to adapt MEA technology to study the mechanism of anesthesia-induced neurotoxicity.

עיבוד של טכנולוגיית מערך Microelectrode לחקר הרדמה-induced Neurotoxicity במוח חזרזיר שלם

Every year, millions of children undergo anesthesia for a multitude of procedures. However, studies in both animals and humans have called into question ...