بدعم من جامعة كنتاكي ، قسم علم الأحياء ، ومكتب للدراسات الجامعية وكلية الآداب والعلوم.

<ol>
	<li>Alexandrowicz, 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=Zur+Kenntnis+des+sympathischen+Nervensystems+der+Crustaceae.">Zur Kenntnis des sympathischen Nervensystems der Crustaceae.</a> Jena Z. Naturw. 45, 395-444 (1909).</li><li>Bungart, D., Dircksen, H., Keller, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+determination+and+distribution+of+the+myotropic+neuropeptide+orcokinin+in+the+nervous+system+of+astacidean+crustaceans.">Quantitative determination and distribution of the myotropic neuropeptide orcokinin in the nervous system of astacidean crustaceans.</a> Peptides. 15, 393-400 (1994).</li><li>Dircksen, H., Burdzik, S., Sauter, A., Keller, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+orcokinins+and+the+novel+octapeptide+orcomyotropin+in+the+hindgut+of+the+crayfish+Orconectes+limosus:+identified+myostimulatory+neuropeptides+originating+totether+in+neurons+of+the+terminal+abdominal+ganglion.">Two orcokinins and the novel octapeptide orcomyotropin in the hindgut of the crayfish Orconectes limosus: identified myostimulatory neuropeptides originating totether in neurons of the terminal abdominal ganglion.</a> J. 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J., Orchard, I., Schmoeckel, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catecholaminergic+neurons+supplying+the+hindgut+of+the+crayfish,+Procambarus+clarkii.">Catecholaminergic neurons supplying the hindgut of the crayfish, Procambarus clarkii.</a> Can. J. Zool. 69, 2778-2785 (1991).</li><li>Mercier, A. J., Orchard, I., TeBrugge, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FMRFamide-like+immunoreactivity+in+the+crayfish+nervous+system.">FMRFamide-like immunoreactivity in the crayfish nervous system.</a> J. exp. Biol. 156, 519-538 (1991).</li><li>Mercier, A. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonergic+modulation+of+crayfish+hindgut.+Biol+Bull.+217(1),+50-64+(2009).">Serotonergic modulation of crayfish hindgut. Biol Bull. 217(1), 50-64 (2009).</a> Biol Bull. 217, 50-64 (2009).</li><li>Shuranova, Z. P., Burmistrov, Y. M., Cooper, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+hundred+years+ago+and+now:+A+short+essay+on+the+study+of+the+crustacean+hindgut.+(Vor+hundert+Jahren+und+nun:+Eine+kurze+Geschichte+von+die+Forschung+des+Hinterdarmes+der+Crustaceen.">A hundred years ago and now: A short essay on the study of the crustacean hindgut. (Vor hundert Jahren und nun: Eine kurze Geschichte von die Forschung des Hinterdarmes der Crustaceen.</a> Crustaceana. 76, 755-670 (2003).</li><li>Shuranova, Z. P., Burmistrov, Y. M., Strawn, J. R., Cooper, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+an+Autonomic+Nervous+System+in+Decapod+Crustaceans.">Evidence for an Autonomic Nervous System in Decapod Crustaceans.</a> International Journal of Zoological Research. 2 (3), 242-283 (2006).</li><li>Siwicki, K. K., Bishop, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+of+proctolinlike+immunoreactivity+in+the+nervsou+systems+of+lobster+and.">Mapping of proctolinlike immunoreactivity in the nervsou systems of lobster and.</a> 243, 435-435 (1986).</li><li>Stemmler, E. A., Cashman, C. R., Messinger, D. I., Gardner, N. P., Dickinson, P. S., Christie, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-mass-resolution+direct-tissue+MALDI-FTMS+reveals+broad+conservation+of+three+neuropeptides+(APSGFLGMRamide,+GYRKPPFNGSIFamide+and+pQDLDHVFLRFamide)+across+members+of+seven+decapod+crustaean+infraorders.">High-mass-resolution direct-tissue MALDI-FTMS reveals broad conservation of three neuropeptides (APSGFLGMRamide, GYRKPPFNGSIFamide and pQDLDHVFLRFamide) across members of seven decapod crustaean infraorders.</a> Peptides. 28, 2104-2115 (2007).</li><li>Wales, W., Sandeman, D. C., Atwood, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+mouthparts+and+gut.">Control of mouthparts and gut.</a> The Biology of Crustacea. 4, 165-191 (1982).</li><li>Webster, S. G., Dircksen, H., Chung, 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=Endocrine+cells+in+the+gut+of+the+shore+crab+Carcinus+maenas+immunoreactive+to+crustacean+hyperglycaemic+hormone+and+its+precursor-related+peptide.">Endocrine cells in the gut of the shore crab Carcinus maenas immunoreactive to crustacean hyperglycaemic hormone and its precursor-related peptide.</a> J. Exp. Biol. 300, 193-205 (2000).</li><li>Winlow, W., Laverack, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+of+hindgut+motility+in+the+lobster+Homarus+gammarus+(L.).+1.+Analysis+of+hindgut+movements+and+receptor+activity.">The control of hindgut motility in the lobster Homarus gammarus (L.). 1. Analysis of hindgut movements and receptor activity.</a> Mar. Behav. Physiol. 1, 1-28 (1972).</li><li>Winlow, W., Laverack, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+of+hindgut+motility+in+the+lobster+Homarus+gammarus+(L.).+2.+Motor+output.">The control of hindgut motility in the lobster Homarus gammarus (L.). 2. Motor output.</a> Mar. Behav. Physiol. 1, 29-47 (1972).</li><li>Winlow, W., Laverack, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+of+hindgut+motility+in+the+lobster+Homarus+gammarus+(L.).+3.+Structure+of+the+sixth+abdominal+ganglion+(6+A.G.)+and+associated+ablation+and+microelectrode+studies.">The control of hindgut motility in the lobster Homarus gammarus (L.). 3. Structure of the sixth abdominal ganglion (6 A.G.) and associated ablation and microelectrode studies.</a> Mar. Behav. Physiol. 1, 93-121 (1972).</li><li>Wrong, A. D., Sammahin, M., Richardson, R., Mercier, A. 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الإعلان عن أي تضارب في المصالح.

التفاصيل الواردة في الفيلم والنص يرتبط وصف الخطوات الرئيسية اللازمة لتسجيل النشاط في المعى المؤخر للجراد في الموقع وكذلك في المختبر. هدف واحد من تقريرنا هو زيادة الوعي لإمكانات هذا في إعداد الطالب وتشغيل مختبرات التحقيق التي تعلم المفاهيم الأساسية في علم وظائف الأعضاء وعلم الصيدلة. ويمكن استخدام هذه الاستعدادات للتحقيق في عدد من الأسئلة التجريبية التي من شأنها أن تؤدي إلى فهم أفضل للوظائف الفسيولوجية للالمعى المؤخر. ولا تزال الآليات الكامنة وراء تنظيم موجات تمعجية وعكس اتجاهها غير معروفة. هي أيضا آليات لزيادة السيطرة على الجهاز الهضمي بأكمله غير مفهومة تماما. مسألة كيفية دمج أعلى مراكز نشاطها مع الإخراج اللاإرادي الذي يتحكم مباشرة في نظام GI يبقى على مساحة مفتوحة من التحقيق (Shuranova وآخرون ، 2006). بالإضافة إلى ذلك ، قدرات osmoregulatory من المعى المؤخر والقشريات وظائف osmoregulation خلال طرح الريش والإجهاد البيئي لم توضح تماما. لا تزال هناك أسئلة كثيرة تنتظر اجابات في هذا الإعداد.

<ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> جراد البحر (Procambarus clarkii). Atchafalaya البيولوجي شركة التموين ، Raceland لوس انجليس. ، الولايات المتحدة الأمريكية. </li><li style=";text-align:right;direction:rtl"> جراد البحر المالحة القياسية : من التعديل فان الحل Harreveld في (1936). (مم) 205 كلوريد الصوديوم ؛ بوكل 5.3 ؛ 13.5 CaCl 2 2H 2 O ؛ 2.45 MgCl 2 6H 2 O (5) ؛ HEPES وتعديلها لدرجة الحموضة 7.4. تصنع مادة السيروتونين ، والدوبامين في الغلوتامات جراد البحر المالحة. ويتم الحصول على جميع المواد الكيميائية من شركة سيغما الكيميائية (سانت لويس ، MO). </li><li style=";text-align:right;direction:rtl"> أدوات تشريح : الجميلة # 5 ملاقط ، مقصات غرامة ، حامل سكين شفرة ، # 26002-20 دبابيس الحشرات (الحصول على جميع العلوم من أدوات الجميلة (الولايات المتحدة) ، وشركة ، 373 - G فخمة بارك محرك ، فوستر سيتي ، كاليفورنيا 94404-1139) </li><li style=";text-align:right;direction:rtl"> Sylgard ذات قاع طبق بتري </li><li style=";text-align:right;direction:rtl"> الأكواب (لعقد المحاليل الكيميائية) </li><li style=";text-align:right;direction:rtl"> يتم تسجيل الإشارات الكهربائية على الخط إلى واجهة 26T PowerLab إلى كمبيوتر (ADInstruments ، كولورادو سبرينغز ، أول أكسيد الكربون ، الولايات المتحدة). نستخدم البرامج القياسية من ADInstruments اسمه تخطيط أو نطاق. <li style=";text-align:right;direction:rtl"> جراب لجسر محول القوة (مطلوبة القرون جسر مختلفة لأنواع مختلفة من محولات القوى). </li><li style=";text-align:right;direction:rtl"> MLT0402 محول القوة (2 غرام) هو الصف البحوث كما هو مبين في الفيديو. MLT 0210 / D (10 ملغ ، 25 غراما) وMLT 500 / A (0 - 500G). هي الدرجات التعليمية محولات (ADInstruments) </li></ol>

physiological experimentation with the crayfish hindgut: a student laboratory exercise

الغرض من التقرير هو وصف تقنيات تشريح لإعداد المعى المؤخر جراد البحر وشرح كيفية جعل التسجيلات الفيزيولوجية مع محول قوة لمراقبة قوة الانكماش. بالإضافة إلى ذلك ، نحن لشرح كيفية مراقبة نشاط تمعجية بصريا ، والتي يمكن استخدامها بوصفها الأحيائي لمختلف الببتيدات والأمينات الاحيائية والعصبية. هذا التحضير هو قابل للطالب في مختبرات الفيزيولوجيا ولما تبديه من المفاهيم الدوائية للطلاب. وقد تم إعداد هذا الاستخدام لأكثر من 100 سنة ، وأنه لا يزال يقدم الكثير كنموذج للتحقيق في توليد وتنظيم الإيقاعات تمعجية واصفا الآليات الكامنة وراء تعديل بهم. والدوائية والمقايسات مستقبلات الفرعية التي كانت بدأت كتابة أكثر من 50 عاما مضت على المعى المؤخر لا تزال تساهم في البحث اليوم. تناسب هذا التحضير قوية لتدريب الطلاب في الفيزيولوجيا وعلم الصيدلة.

Watch this Scientific Journal Video about Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise at JoVE.com

Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise

في هذا التقرير أن نبرهن التقنيات التي يمكن استخدامها للتحقيق في علم الأحياء من جراد البحر المعى المؤخر. نعرض كيفية تشريح البطن جراد البحر ودراسة علم التشريح المرتبطة بها ، وعلم وظائف الأعضاء تعديل النشاط. ويتم قياس نشاط تمعجية وقوة تقلصات باستخدام محول القوة.

الفسيولوجية التجريب مع المعى المؤخر جراد البحر : تمرين الطلاب مختبر

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

physiological-experimentation-with-crayfish-hindgut-student

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Neuroscience

15.7K Views.  University of Kentucky. في هذا التقرير أن نبرهن التقنيات التي يمكن استخدامها للتحقيق في علم الأحياء من جراد البحر المعى المؤخر. نعرض كيفية تشريح البطن جراد البحر ودراسة علم التشريح المرتبطة بها ، وعلم وظائف الأعضاء تعديل النشاط. ويتم قياس نشاط تمعجية وقوة تقلصات باستخدام محول القوة.

Video: Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise

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.

تسجيل الاستجابات السلوكية للتفكير في جراد البحر

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.

إجراء دراسة لإعادة Footshock الناتج عن السعي من الكوكايين في فئران التجارب

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.

الفسيولوجية تسجيلات NMJs المخرجات العالية والمنخفضة على العضلة الباسطة جراد البحر الساق

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.

الإمكانيات الغشاء ، الردود متشابك ، الدارات العصبية ، تعديل العمليات العصبية والأنسجة العضلية باستخدام جراد البحر : تمارين مختبر طالبة

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 ...

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

The primary purpose of this experiment is to demonstrate primary sensory neurons conveying information of joint movements and positions as proprioceptive information for an animal. An additional objective of this experiment is to learn anatomy of the preparation by staining, dissection and viewing of neurons and sensory structures under a dissecting microscope. This is performed by using basic neurophysiological equipment to record the electrical activity from a joint receptor organ and staining techniques. The muscle receptor organ (MRO) system in the crayfish is analogous to the intrafusal muscle spindle in mammals, which aids in serving as a comparative model that is more readily accessible for electrophysiological recordings. In addition, these are identifiable sensory neurons among preparations. The preparation is viable in a minimal saline for hours which is amenable for student laboratory exercises. The MRO is also susceptible to neuromodulation which encourages intriguing questions in the sites of modulatory action and integration of dynamic signals of movements and static position along with a gain that can be changed in the system.

The primary purpose of this experiment is to understand how primary sensory neurons convey information of joint movements and positions as proprioceptive information for an animal. An additional objective of this report is present the anatomy of the preparation by dissection and viewing of neurons under a dissecting microscope.

هيئات مستقبلات العضلات في البطن جراد البحر : تمرين الطلاب في مختبر استقبال الحس العميق

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

VisualEyes: A Modular Software System for Oculomotor Experimentation

Eye movement studies have provided a strong foundation forming an understanding of how the brain acquires visual information in both the normal and dysfunctional brain.1 However, development of a platform to stimulate and store eye movements can require substantial programming, time and costs. Many systems do not offer the flexibility to program numerous stimuli for a variety of experimental needs. However, the VisualEyes System has a flexible architecture, allowing the operator to choose any background and foreground stimulus, program one or two screens for tandem or opposing eye movements and stimulate the left and right eye independently. This system can significantly reduce the programming development time needed to conduct an oculomotor study. The VisualEyes System will be discussed in three parts: 1) the oculomotor recording device to acquire eye movement responses, 2) the VisualEyes software written in LabView, to generate an array of stimuli and store responses as text files and 3) offline data analysis. Eye movements can be recorded by several types of instrumentation such as: a limbus tracking system, a sclera search coil, or a video image system. Typical eye movement stimuli such as saccadic steps, vergent ramps and vergent steps with the corresponding responses will be shown. In this video report, we demonstrate the flexibility of a system to create numerous visual stimuli and record eye movements that can be utilized by basic scientists and clinicians to study healthy as well as clinical populations.

Neural control and cognitive processes can be studied through eye movements. The VisualEyes software allows an operator to program stimuli on two computer screens independently using a simple, custom scripting language. The system can stimulate tandem eye movements (saccades and smooth pursuit) or opposing eye movements (vergence) or any combination.

VisualEyes: نظام وحدات البرمجيات لالمحرك للعين التجريب

Eye movement studies have provided a strong foundation forming an understanding of how the brain acquires visual information in both the normal and ...

Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex behaviors such as food choice, predator, conspecific and mate recognition and other socially relevant cues. Olfactory receptor neurons (ORNs) are located in the dorsal part of the nasal cavity embedded in the olfactory epithelium. These bipolar neurons send an axon to the olfactory bulb (see Fig. 1, Reisert &amp; Zhao1, originally published in the Journal of General Physiology) and extend a single dendrite to the epithelial border from where cilia radiate into the mucus that covers the olfactory epithelium. The cilia contain the signal transduction machinery that ultimately leads to excitatory current influx through the ciliary transduction channels, a cyclic nucleotide-gated (CNG) channel and a Ca2+-activated Cl- channel (Fig. 1). The ensuing depolarization triggers action potential generation at the cell body2-4.
In this video we describe the use of the "suction pipette technique" to record odorant-induced responses from ORNs. This method was originally developed to record from rod photoreceptors5 and a variant of this method can be found at jove.com modified to record from mouse cone photoreceptors6. The suction pipette technique was later adapted to also record from ORNs7,8. Briefly, following dissociation of the olfactory epithelium and cell isolation, the entire cell body of an ORN is sucked into the tip of a recording pipette. The dendrite and the cilia remain exposed to the bath solution and thus accessible to solution changes to enable e.g. odorant or pharmacological blocker application. In this configuration, no access to the intracellular environment is gained (no whole-cell voltage clamp) and the intracellular voltage remains free to vary. This allows the simultaneous recording of the slow receptor current that originates at the cilia and fast action potentials fired by the cell body9. The difference in kinetics between these two signals allows them to be separated using different filter settings. This technique can be used on any wild type or knockout mouse or to record selectively from ORNs that also express GFP to label specific subsets of ORNs, e.g. expressing a given odorant receptor or ion channel.

Olfactory receptor neurons (ORNs) convert odor signals first into a receptor current that in turn triggers action potentials that are conveyed to second order neurons in the olfactory bulb. Here we describe the suction pipette technique to record simultaneously the odorant-induced receptor current and action potentials from mouse ORNs.

الرائحة التي يسببها الردود المسجلة من الخلايا العصبية مستقبلات الشم باستخدام تقنية شفط الماصة

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex ...

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.

استقبال الحس العميق والتوتر المستقبلات في اطراف سرطان البحر: تمارين مختبر طالبة

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.

قياس الأحماض الدهنية عن طريق الفم العتبات، الدهون الإدراك، الأغذية الدهنية الميل، والحليمات الكثافة في البشر

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

Lineage-reprogramming of Pericyte-derived Cells of the Adult Human Brain into Induced Neurons

Direct lineage-reprogramming of non-neuronal cells into induced neurons (iNs) may provide insights into the molecular mechanisms underlying neurogenesis and enable new strategies for in vitro modeling or repairing the diseased brain. Identifying brain-resident non-neuronal cell types amenable to direct conversion into iNs might allow for launching such an approach in situ, i.e. within the damaged brain tissue. Here we describe a protocol developed in the attempt of identifying cells derived from the adult human brain that fulfill this premise. This protocol involves: (1) the culturing of human cells from the cerebral cortex obtained from adult human brain biopsies; (2) the in vitro expansion (approximately requiring 2-4 weeks) and characterization of the culture by immunocytochemistry and flow cytometry; (3) the enrichment by fluorescence-activated cell sorting (FACS) using anti-PDGF receptor-&#946; and anti-CD146 antibodies; (4) the retrovirus-mediated transduction with the neurogenic transcription factors sox2 and ascl1; (5) and finally the characterization of the resultant pericyte-derived induced neurons (PdiNs) by immunocytochemistry (14 days to 8 weeks following retroviral transduction). At this stage, iNs can be probed for their electrical properties by patch-clamp recording. This protocol provides a highly reproducible procedure for the in vitro lineage conversion of brain-resident pericytes into functional human iNs.

Targeting brain-resident cells for direct lineage-reprogramming offers new perspectives for brain repair. Here we describe a protocol of how to prepare cultures enriched for brain-resident pericytes from the adult human cerebral cortex and convert these into induced neurons by retrovirus-mediated expression of the transcription factors Sox2 and Ascl1.

النسب-إعادة برمجة الخلايا المشتقة من خلية حوطية الدماغ البشري الكبار في الخلايا العصبية المستحثة

Direct lineage-reprogramming of non-neuronal cells into induced neurons (iNs) may provide insights into the molecular mechanisms underlying neurogenesis ...