Name: training rats to voluntarily dive underwater: investigations of the mammalian diving response
Uploaded: 2014-11-12T15:00:00.000+00:00
Duration: 11 min 56 s
Description: Watch this Scientific Journal Video about Training Rats to Voluntarily Dive Underwater: Investigations of the Mammalian Diving Response at JoVE.com

Araştırma Araştırma ve Sponsorlu Programları Ortabatı Üniversitesi Ofisi tarafından finanse edilerek desteklenmiştir. Ayrıca Ortabatı Üniversitesi Hayvan Tesisi ve Erik Warren teşekkürler.

<ol>
	<li>Butler, P. J., Jones, D. R., Lowenstein, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Advances in Comparative Physiology and Biochemistry. 8, 179-364 (1982).</li><li>Butler, P. J., Jones, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiology+of+diving+birds+and+mammals).">Physiology of diving birds and mammals).</a> Physiol. Rev. 77, 837-899 (1997).</li><li>Butler, 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=Metabolic+regulation+in+diving+birds+and+mammals.">Metabolic regulation in diving birds and mammals.</a> Resp. Physiol. Neurobiol. 141, 297-315 (2004).</li><li>Foster, G. E., Sheel, A. 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Physiol. 26, 395-401 (1976).</li><li>Fahlman, A., Bostrom, B. L., Dillon, K. H., Jones, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetic+component+of+the+forced+diving+bradycardia+response+in+mammals.">The genetic component of the forced diving bradycardia response in mammals.</a> Front. Physiol. 2, 1-7 (2011).</li><li>Blix, A. S., Folkow, B., Shepher, J. T., Abboud, F. 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> Handbook of Physiology. , 917-944 (1984).</li><li>Kooyman, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diverse+Divers.">Diverse Divers.</a> 200, (1989).</li><li>MacArthur, R. A., Karpan, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+rates+of+muskrats+diving+under+simulated+field+conditions:+persistence+of+the+bradycardia+response+and+factors+modifying+its+expression.">Heart rates of muskrats diving under simulated field conditions: persistence of the bradycardia response and factors modifying its expression.</a> Can. J. Zool. 67, 1783-1792 (1989).</li><li>McCulloch, P. F., Jones, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+influences+on+diving+bradycardia+in+muskrats+(Ondatra+zibethicus).">Cortical influences on diving bradycardia in muskrats (Ondatra zibethicus).</a> Physiol. Zool. 63, 1098-1117 (1990).</li><li>McCulloch, P. F., Dinovo, K. M., Connolly, T. 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+cardiovascular+and+endocrine+responses+to+voluntary+and+forced+diving+in+trained+and+untrained+rats.">The cardiovascular and endocrine responses to voluntary and forced diving in trained and untrained rats.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 298, 224-234 (2010).</li><li>Panneton, W. M., Gan, Q., Dahms, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiorespiratory+and+neural+consequences+of+rats+brought+past+their+aerobic+dive+limit.">Cardiorespiratory and neural consequences of rats brought past their aerobic dive limit.</a> J. Appl. Physiol. 109, 1256-1269 (2010).</li><li>Chotiyanonta, J. S., DiNovo, K. M., McCulloch, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bilateral+sectioning+of+the+anterior+ethmoidal+nerves+does+not+eliminate+the+diving+response+in+voluntarily+diving+rats.">Bilateral sectioning of the anterior ethmoidal nerves does not eliminate the diving response in voluntarily diving rats.</a> Physiol. Reports. 1, (2013).</li><li>Panneton, W. M., Anch, A. M., Panneton, W. M., Gan, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parasympathetic+preganglionic+cardiac+motoneurons+labeled+after+voluntary+diving.">Parasympathetic preganglionic cardiac motoneurons labeled after voluntary diving.</a> Front. Physiol. 5, 1-10 (2014).</li><li>McCulloch, P. F., Ollenberger, G. P., Bekar, L. K., West, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trigeminal+and+chemoreceptor+contributions+to+bradycardia+during+voluntary+dives+in+rats.">Trigeminal and chemoreceptor contributions to bradycardia during voluntary dives in rats.</a> Am. J. Physiol. Regul. Integr. Comp. Physiol. 273, 814-822 (1997).</li><li>Ollenberger, G. P., Matte, G., Wilkinson, A. A., West, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relative+distribution+of+blood+flow+in+rats+during+surface+and+submerged+swimming.">Relative distribution of blood flow in rats during surface and submerged swimming.</a> Comp. Biochem. Physiol. A. 119, 271-277 (1998).</li><li>Ollenberger, G. P., West, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distribution+of+regional+cerebral+blood+flow+in+voluntarily+diving+rats.">Distribution of regional cerebral blood flow in voluntarily diving rats.</a> J. Exp. Biol. 201, 549-558 (1998).</li><li>McCulloch, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+the+trigeminal+medullary+dorsal+horn+during+voluntary+diving+in+rats.">Activation of the trigeminal medullary dorsal horn during voluntary diving in rats.</a> Brain Res. 1051, 194-198 (2005).</li><li>Panneton, W. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+brainstem+neurons+by+underwater+diving+in+the+rat.">Activation of brainstem neurons by underwater diving in the rat.</a> Front. Physiol. 3, 1-13 (2012).</li><li>McCulloch, P. F., Panneton, W. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+brainstem+catecholaminergic+neurons+during+voluntary+diving+in+rats.">Activation of brainstem catecholaminergic neurons during voluntary diving in rats.</a> Brain Res. 984, 42-53 (2003).</li><li>McCulloch, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Globosa+neurons:+a+distinct+subgroup+of+noradrenergic+neurons+in+the+caudal+pons+of+rats.">Globosa neurons: a distinct subgroup of noradrenergic neurons in the caudal pons of rats.</a> Brain Res. 964, 164-167 (2003).</li><li>Panneton, W. 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+mammalian+diving+response:+an+enigmatic+reflex+to+preserve+life.">The mammalian diving response: an enigmatic reflex to preserve life.</a> Physiology. 28, 284-297 (2013).</li></ol>

Yazar Stoelting Company için bir danışman ve ticari amaçlar için kendilerine genel tasarım ve McCulloch Dalış Tank Maze özelliklerini sağladı.

Onların vahşi formu can Sıçanlar ve yarı-sucul ortamlarda istismar yapmak ve yiyecek 6 aramak olurken olacak sık sık sualtı dalış. Böylece fareler çok kolay gönüllü sualtı dalış için eğitilmiş olabilir bu çok şaşırtıcı değildir. Tarif edilen eğitim prosedürler çoğu erişkin sıçan beyin atlaslarında kullanılan bir vücut büyüklüğü (~ 300 g) yeni sütten fareler getirecek 6 haftaya kadar sürebilir. Bu eğitimli hayvanların Böylece beyinleri daha kolay karşılaştırılabilir bu atlaslarında tanımlanan anatomik yapılara olacaktır. Start alanında konulduktan sonra en ratlar 20 saniye içerisinde kendi sualtı yüzmek başlayacak. Ancak, bazen bir sıçan gönüllü dalışa başlamadan önce 5 dakika veya daha fazla sürecektir. Bu sualtı tüneli içine sıçan zorlamak için bu aşamada cazip olsa da, bu genelde olumsuz bir deneyim ile suyun ilişkilendirme gelen fareleri önlemek için kaçınılmalıdır. Kemirgenler, başlatılmış inatçı olabilir ve olabilir olabilirmüttefiki eğitim oturumları sırasında dalış reddediyorum, ama onlar su çıkmak için tek yolu labirent aracılığıyla dalış tamamlayarak tarafından fark bir kez, genellikle yakında start alanında yatırıldıktan sonra dalış başlatır. Tekrarlayan ve metodik eğitim prosedürlerin kritik yönü onlar sıçan yaşadığı stresi azaltmak için böyle bir şekilde yürütülen olmasıdır. Sıçanlar özellikle denemeleri arasında bitirme alanında ise, çevrelerini keşfetmek izin, daha da stresi azaltmak gibi görünüyor. Sıçanlar sık ​​sık yüzmek için suya tekrar girin ve / veya hala bitiş alanı platformda otururken kendi baş sualtı batığın olacaktır. Bu fareler doğal su caydırıcı olmadığını göstermektedir. Ayrıca, sıçanlar yüzme veya dalış, ya da daha az fekal pelet 17 bitiş alanında bekleyen, günlük eğitim sonuçlarına sırasında fekal pelet üretmek için nadir değildir iken. Genel olarak daha az fareler eğitim sırasında olduğu vurguladı, daha az pellets onlar üretecek. Üretilen herhangi pislikleri nispeten temiz su tutmaya kısa sürede su veya bitiş alanından kaldırılır. Sıçanlar bazen kanlı burun alabilirsiniz burun pasajda suyun içine çekilmesinin bir sonucu olabilir yüzme ve dalış, süre. Kanın görünümü burun mukozasına olan ozmotik streslere bağlı olabilir. Çalışmalar arasında finiş alanında ise fareler kendilerini damat olacak. Burun bir sonucu kan gibi bir eğitim oturumu sırasında, özellikle göz çevresinde, sıçanlar hafif kırmızımsı bir renk vererek, fareler baş ve burun üzerinde dağıtılabilir alabilirsiniz. Ayrıca, arada bir sıçan dalış stresli olabilir ve / veya olumsuz dalış deneyimleri var (etrafında dönen ve dalış sualtı (5.4.3 sonra not görürken kaybolmadan tarafından, yani. On) oluşmasını engellemek için nasıl). Bu sıçanlarda porfirin bir stres RESPONS sinyalizasyon, onların gözlerinin köşelerinde görünebilirör. Dalış tankın büyüklüğü belli bir dereceye kadar oda gereksinimlerini belirlemek olacaktır. Tarif tankı sıçanlar 5 m uzunluğunda pleksiglas sayesinde suyun yüzeyinin altında 2-3 cm yüzmek için tasarlanmıştır 10-15 sn 17,19,24,26,27 bir sualtı yüzme süresini vermek labirent. Bir deney daha uzun bir süre dalıştan tepkilerini ölçmek için tasarlanmış olmalıdır, ya da sualtı bir derin dalıştan, tankı yeniden dizayn edilmesi gerekebilir. Oda gereksinimleri sonra da yeniden tasarlanmış dalış tank boyutlarına sığacak şekilde değişebilir. Prosedür kullanılabilir odasında hiçbir zemin drenaj yoksa, tanktan su gibi, sonra uygun bir şekilde başka bir yerde boşaltılabilir bir 60 gal çöp can, gibi, büyük bir kap içinde toplanabilir. Fos teknik, ayrıca, dalış tepkisinin beyin devresinin bir parçası olan nöronlar belirlemek ve karakterize etmek için, diğer sinir tespit yöntemleri ile kullanılabilir. Örneğin,Gönüllü sırasında aktive olan yan A7 alanı 26,27 mesafede olan ve globosa&#39;nın nöronlar, tirozin hidroksilaz boyama ile bağlantılı Fos algılama A1, C1, A2, A5 katekolaminerjik nöronlar ve alt coeruleus alanlar (26 Şekil 6) belirlemiştir dalış. Ayrıca geriye iz Kolera toksini ile bağlantılı Fos algılama gönüllü dalış 20 sırasında aktive olan çekirdek kuşkulu dış oluşum içinde kardiyak vagus nörotoksinlerini burada hücre gövdelerini belirlemiştir. Dalış kardiyorespiratorik tepkilerin merkezi sinir entegrasyonu incelenmesi nedenlerden 6,8,28 bir dizi için önemlidir. Dalış tepki uzun süre boyunca nefes almadan batık sualtı kalmaya hayvanlar dahil olmak üzere insanları, sağlar. Dalış yanıt beyin sapı homeostatik kontrol fonksiyonel bir yeniden temsil eder ve en güçlü pa birini gösterirHayvanlarda görülen otonomik refleksleri tterns. Dalış yanıt da trigemino-kardiyak refleks parçası, nazofarengeal refleksi ve / veya ani bebek ölümü sendromu gibi insanlarda önemli klinik olabilir. Son olarak, sıçanların beyin sapı içinde var nöronal devrenin bir anlayış afferent sinyalleri temel beyin sapı otonomik refleksler nasıl değiştirebileceğiniz kortikal belirlemenize yardımcı olacaktır. Bu düşünceler tüm memeli dalış tepkisinin merkezi yönlerini çalışma doğal olarak değerli ve ilginç olun. Açıklanan prosedürler kullanılarak gönüllü sualtı zorla daldı hayvanların kullanımı olacak daha memeli dalış tepkisinin merkezi yönlerini daha iyi soruşturma sağlayacak dalış fareleri yetiştirmektir. Tarif 1 olarak eğitim prosedürleri) MSS stres devre aktivasyonunu azaltan ve dış ödüller kullanılmaz çünkü 2) CNS ödül devreyi aktif değil çünkü bu.

The diving response consists of a suite of autonomic reflexes seen in animals of all vertebrate classes 1. In response to submersion under water, this reflexly-induced response consists of apnea, bradycardia and an alteration of blood flow that maintains flow to the heart, brain and exercising muscles while limiting flow to viscera and non-exercising muscles 2. Many of the metabolic and cardiorespiratory aspects of the mammalian diving response have been well investigated 2,3, including those in humans 4,5. However, what has been relatively lacking, until recently, is investigation of the central integrative aspects of the diving response. What happens within the brainstem, and what is the neuronal step-by-step pathway, that connects afferent inputs to efferent outputs during this autonomic reflex? Answering these questions will require an appropriate animal model 6. An adage in comparative physiology, the Krogh principle 7, is that for every research question there is some animal of choice on which the problem can be most conveniently studied. A most appropriate animal for studying the central aspects of the diving response is the rat 6,8. In large part this is due to the fact that the brains of rats have been very well characterized, both anatomically and functionally, and many rat brain atlases are available 6. Additionally the rat is particularly useful in cardiorespiratory research, because the physiology of the rat is well known across all major organ systems, and the rat is well regarded as an animal model in systems biology 6. Finally, the laboratory rat is the domesticated version of the wild Rattus norvegicus, an animal that routinely swims and dives underwater 6. Based on these considerations, the rat is a good choice for studies investigating the central aspects of the mammalian diving response. In comparison, using marine animals to investigate the central aspects of the mammalian diving response would be much more difficult. This is due in large part to marine animals having comparatively large and non-uniformly sized brains, and the relative difficulty and high cost of housing these animals.
Rats have previously been used to investigate many aspects of the mammalian diving response, primarily in situations involving forced underwater submergence 9-12. However many studies in marine and aquatic animals have shown that there can be a differential response to diving based upon whether the submergence was forced or voluntary 2,13. Diving animals may show an extremely intense bradycardia during forced diving but a much less intense bradycardia during voluntary diving. The &#8220;stress&#8221; of forced submergence can significantly change the cardiovascular responses of diving in many animals 14. Small rodents such as muskrats also show a more intense diving response during forced submergence than during voluntary diving 15,16. Thus, if rats are to be used to investigate the central aspects of the mammalian diving response, investigators should be aware that a rat forcibly submerged underwater may produce a response different from that of a voluntarily diving rat. 
The goal of this article is to provide detailed instructions on how to train rats to voluntarily dive underwater. These procedures are not technically difficult, but can be time-consuming. The training is conducted in such a way as to reduce the stressfulness that could otherwise be associated with forced underwater submergence. This voluntary diving technique should minimize activation of central stress pathways and thus better allow investigation of the central aspects of the diving response. By itself, training rats to voluntarily dive underwater generates no data that can be used to investigate the central aspects of the mammalian diving response. Therefore examples of how voluntarily diving rats have been used in conjunction with other physiological and neuroanatomical research techniques, and how the basic dive training may need to be modified to accommodate theses other techniques, are also provided.

<table><tbody><tr><td>McCulloch Diving Tank Maze</td><td>Stoelting Company</td><td>60139</td><td></td></tr><tr><td>1 inch internal diameter tubing&amp;nbsp;</td><td>Fisher</td><td>14-169-63</td><td>Used to fill or drain tank</td></tr><tr><td>Plexiglas rodent restraint device (Economy flat bottomed restrainer)</td><td>Braintree</td><td>FB-M/L&amp;nbsp;</td><td>For forced dives</td></tr><tr><td>Telemetric transmitters&amp;nbsp;</td><td>DSI</td><td>Model PA-C40 (270-0040-008)</td><td>Used to transmit pulsatile arterial blood pressure</td></tr><tr><td>Hand-held antenna wand</td><td>DSI</td><td>Model RLA 3000 (272-5007)</td><td>Used to ensure radio antenna is near to transmitter while rat is negotiating underwater maze</td></tr><tr><td>Intramedic PE50, 0.023&quot; ID</td><td>Fisher</td><td>14-170-12B</td><td>Used as trailing arterial cannula</td></tr><tr><td>N95 mask - Moldex #2300N Series</td><td>Fisher</td><td>19-003-246D</td><td>Used to limit inhalation of rat allergens</td></tr></tbody></table>

training rats to voluntarily dive underwater: investigations of the mammalian diving response

Underwater submergence produces autonomic changes that are observed in virtually all diving animals. This reflexly-induced response consists of apnea, a parasympathetically-induced bradycardia and a sympathetically-induced alteration of vascular resistance that maintains blood flow to the heart, brain and exercising muscles. While many of the metabolic and cardiorespiratory aspects of the diving response have been studied in marine animals, investigations of the central integrative aspects of this brainstem reflex have been relatively lacking. Because the physiology and neuroanatomy of the rat are well characterized, the rat can be used to help ascertain the central pathways of the mammalian diving response. Detailed instructions are provided on how to train rats to swim and voluntarily dive underwater through a 5 m long Plexiglas maze. Considerations regarding tank design and procedure room requirements are also given. The behavioral training is conducted in such a way as to reduce the stressfulness that could otherwise be associated with forced underwater submergence, thus minimizing activation of central stress pathways. The training procedures are not technically difficult, but they can be time-consuming. Since behavioral training of animals can only provide a model to be used with other experimental techniques, examples of how voluntarily diving rats have been used in conjunction with other physiological and neuroanatomical research techniques, and how the basic training procedures may need to be modified to accommodate these techniques, are also provided. These experiments show that voluntarily diving rats exhibit the same cardiorespiratory changes typically seen in other diving animals. The ease with which rats can be trained to voluntarily dive underwater, and the already available data from rats collected in other neurophysiological studies, makes voluntarily diving rats a good behavioral model to be used in studies investigating the central aspects of the mammalian diving response.

Açıklanan yüzme ve dalış eğitimi prosedürlerin başarıyla tamamlanması su altında dalış yaparken sıçanların yaşadığı stresi azaltabilir. Kan kortikosteron seviyeleri tekrarlanan günlük eğitim gönüllü dalış ile ilişkili stressfulness azaldığını göstermektedir, ve eğitilmiş fareler insan (Şekil 2; 17) tarafından günlük olarak işlenen daha dalış fazla stresli bulabilirsiniz. Tersine, dalış protokolde eğitimli sıçan gönüllü dalış stresli (Şekil 2; 17) bulabilirsiniz. Ayrıca, her ikisi de eğitimli ve eğitimsiz sıçanlar en stresli (; 17 Şekil 2) olması için dalış zorunda bulabilirsiniz. Ve sondaki kanüller (Şekil 4; 21-23); yüzme, gönüllü dalış ve zorla dalış sıçanların kardiyovasküler tepkiler implante telemetri cihazları (8,17-20 Şekil 3) kullanılarak kaydedilmiştir. Hemen gönüllü üzerindebatma ve tek bir ritmi içinde, kalp atım hızı düşer% 78 oranında ve% 25 17 arteriyel kan basıncı azalır demek. Bu sonuçlar gönüllü dalış sıçanlar genellikle diğer dalış hayvanlarda görülen aynı kardiyorespiratori değişiklikleri sergilemek olduğunu göstermektedir. Ve gönüllü dalış sırasında serebral kan akımı 23 olmak üzere kardiyak debinin 22, dağılımını belirlemek için; Firar arteriyel kanül gönüllü dalış (21 Şekil 4) ile ilişkili bradikardi ortadan muskarinik antagonist atropin, enjekte etmek için kullanılmıştır. Firar kanüller ayrıca 18 batık ise sıçan arteriyel hipoksemi ve hiperkapni artan görmezden olduğunu göstermek için kullanılan ve edildiğini önceden varolan chemoreceptor sürücüyü gönüllü dalış 21 kardiyovasküler tepkiler üzerine herhangi bir etkisi yoktur. Lamina I ve ventral medüller dorsal boynuz II (MDH) ifade Fos durin içinde Nöronlarg gönüllü dalış, ve bu nöronlar dalış yanıt (; 24 Şekil 5) ilk beyin sapı afferent rölesi teşkil edebilir. Böyle kaudal pressör alanı (CPA) gibi önemli beyin sapı kardiyorespiratuvar kontrol alanları, nükleus traktus solitaries (NTS), rostran ventrolateralinde medulla (RVLM) ve peribrachial bölgeler, tüm gösteri yüzme 25 ile karşılaştırıldığında gönüllü dalış sırasında Fos etiketleme arttı. Uzun süreli dalışlar 18 zorla sonra beyin sapı kemosensitif bölgelerinde Nöronlar Fos ifade eder. <img alt="Şekil 1," fo:content-width="4in" src="/files/ftp_upload/52093/52093fig1highres.jpg" width="400" /> Şekil 1:. Dalış Tank şematik bir pleksiglas tankı (100 x 60 x 15 cm), beş 1 m uzunluğunda kanallardan oluşan basit bir labirent oluşturmak için kullanılmıştır. Tank, 30 ° C musluk suyu ile doldurulmuş ve fareler ilk negotiat şekilde eğitildiFinish Area (sol üst) Başlat Area, su yüzeyinde yüzerek e labirenti (sağ alt). Farelere daha sonra labirent aracılığıyla dalış eğitilmiştir, su yüzeyinin altında 2-3 cm yerleştirilmiş yatay pleksiglas parçaları ile sualtı tuttu. [Bu rakam 26 dan modifiye edilmiştir] <img alt="Şekil 2," fo:content-width="5in" src="/files/ftp_upload/52093/52093fig2highres.jpg" width="500" /> Şekil 2: Kortikosteron ölçümleri. Kan sıçanlar 10 dakika / sıçan yüzmek ve dalış (eğitimli) için eğitilmiş gün (Kulplu), ve sıçanlar için ele, onların kafeslerinde (Naive) sol sıçanların corticosterone konsantrasyonlarını (ortalama ± SH) ölçmek için kullanılan sıçan kuyruğu damarlar berabere hiçbir yüzmek ya da dalış eğitimi (Eğitimsiz) aldı. Eğitimli fareler kendi yüzmeye eğitimi (Sol barlar set) tamamlandıktan sonra corticosterone ölçüldü, sonra eğitilmiş fareler kendi gönüllü div tamamlamıştıe eğitim (barlar Merkezi seti), ve eğitimli sonra, sıçanlar (Sağ barlar set) zorla dalış eğitimini tamamladı. 1 değeri Naïve anlamlı ölçüde daha büyük olduğunu gösterir; 2 değeri işlenmiş anlamlı ölçüde daha büyük olduğunu gösterir; 3 değeri Eğitimli anlamlı ölçüde daha büyük olduğunu gösterir; * Zorunlu dalış sırasında eğitimli fareler değer gönüllü dalış sırasında önemli ölçüde fazla olduğunu gösterir. [Bu rakam 17 ile modifiye edilmiştir] <img alt="Şekil 3," fo:content-width="5in" src="/files/ftp_upload/52093/52093fig3highres.jpg" width="500" /> Şekil 3: telemetrik vericilerinden gelen arteriyel kan basıncı izleri labirent aracılığıyla yüzmek için eğitilmiş fareler ve dalıştan (sol sütun), gönüllü dalış (orta sütun), ve zorunlu dalış (sağ sütun) yüzme sırasında pulsatil arteriyel kan basıncını gösteren Ham veri izleri. Öğ yoktu sıçanların (alt sıra) vening prosedür (üst satır). Su yüzeyinde yüzme böyle bir kardiyovasküler değişikliklere neden ise dalış sualtı (gönüllü ve zorla batıklık her ikisi de), acil bradikardi ve arteriyel basınçta yavaş başlangıçlı artış üretti. Izleri altında Barlar daldınlma dönemi işaret etmektedir. Telemetrik sinyalin kaybolduğu zaman iz tatili dönemi işaret etmektedir. [Bu rakam 17 ile modifiye edilmiştir] <img alt="Şekil 4," fo:content-width="5in" src="/files/ftp_upload/52093/52093fig4highres.jpg" width="500" /> Şekil 4: Antropin dalış bradikardi ortadan kaldırır. Atropin ön işlemden sonra, pulsatil kan önce gönüllü dalma sıçanlarda (A) basınç ve (B) &#39;nin orijinal kayıtları. İzler bir arka arteriyel kanül kullanılarak elde edilmiştir. Atropin ön tedaviden önce, tansiyon batma üzerine biraz azalmış, ama ttavuk dalış kalan dalış öncesi daha fazla yükselmiştir. Kalp hızı komşu nabız basıncı aralıklarla belirlendi. Suya batma üzerine dalış süresince devam etti acil ve önemli bradikardi oldu. Atropin tedavi öncesi ile parasempatik ablukasının sonra bradikardi elendi. Dalış sırasında arter basıncında bir artış oldu. Iz altındaki bar daldınlma süreyi gösterir. [McCulloch, yayınlanmamış] <img alt="Şekil 5," fo:content-width="6in" src="/files/ftp_upload/52093/52093fig5highres.jpg" width="600" /> Şekil 5: MDH içinde Fos etiketleme. Trigeminal medüller dorsal boynuz (MDH) ve sualtı dalış için eğitilmiş sıçanlarda spinal trigeminal sistem (sp5) fotomikrografları. Tekrarlanarak hiçbir Fos etiketleme yok değil dalış bir kontrol sıçan (A). (B) yüzme Sıçan çok yanar varSP5 içinde tle Fos MDH etiket (büyük ok) ya da paratrigeminal çekirdek (küçük ok). (C) dalış sıçan daha Fos etiketleme MDH (büyük ok) hem ventral var ve paratrigeminal nükleus (küçük oklar) olarak karşılaştırıldığında yüzme ve kontrol sıçan. Panel (a) olarak kaydedin panel ac rostral-kaudal konumunu gösterir. Panel C Ölçek çubuğu 100 mikron. [Bu rakam 24 değişiklikler yapılarak elde edilmiş] <img alt="Şekil 6," fo:content-width="5in" src="/files/ftp_upload/52093/52093fig6highres.jpg" width="500" /> Şekil 6: dalış sıçanların Aktif katekolaminerjik nöronlar. Fotomikrograflar olmayan bir dalış kontrol faresi (A, C, ve E) ve gönüllü dalış farede (B, D ve F) medullas göstermektedir. Beyin dokusu immunohistologically hem Fos An için işlendid tirozin hidroksilaz (TH), kahverengi TH somas ve siyah Fos çekirdekleri üreten. Oklar Fos + TH çift etiketli nöronlar tespit ederken Açık sıra, tek etiketli TH-pozitif nöronlar tespit. A1 nöronlar A ve B C1-nöronlarda tanımlanan C tanımlanır ve D A5 nöronlar E ve F Daha Fos ve A1, C1 görülen çift etiketli TH tespit edilir ve dalma sıçan daha A5 bölgeleri olmayan dalış kontrol sıçanlarda. E Kalibrasyon çubuğu paneller AF için, ve 250 mm. F içerlek Kalibrasyon çubuğu tüm ilavelerde için, ve 50 mm. [Bu rakam 26 dan modifiye edilmiştir]

Watch this Scientific Journal Video about Training Rats to Voluntarily Dive Underwater: Investigations of the Mammalian Diving Response at JoVE.com

Training Rats to Voluntarily Dive Underwater: Investigations of the Mammalian Diving Response

Detailed instructions are provided on how to train rats to voluntarily dive underwater through a 5 m long Plexiglas maze. Because the brains of rats have been very well characterized, voluntarily diving rats may help elucidate the central pathways of the mammalian diving response.

Gönüllü Dalış Sualtı eğitim Rats: Memeli Dalış Tepki Araştırmalar

Underwater submergence produces autonomic changes that are observed in virtually all diving animals. This reflexly-induced response consists of apnea, a ...

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Nanyang Technological University

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JoVE Journal

Behavior

12.2K Görüntüleme.  Midwestern University. Detailed instructions are provided on how to train rats to voluntarily dive underwater through a 5 m long Plexiglas maze. Because the brains of rats have been very well characterized, voluntarily diving rats may help elucidate the central pathways of the mammalian diving response.

Video: Training Rats to Voluntarily Dive Underwater: Investigations of the Mammalian Diving Response

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to the mouse becoming increasingly important. But researchers found that some strains of mutant mice were prone to problems like passively floating or diving when they were tested in the Morris water maze11. This was unsurprising considering their natural habitat; rats swim naturally (classically, the "sewer rat"), whereas mice evolved in the dry areas of central Asia.
To overcome these problems, it was considered whether shallow water would be a sufficient stimulus to provide escape motivation for mice. This would also avoid the problems of drying the small creatures with a towel and then putting them in a heated recovery chamber to avoid hypothermia, which is a much more serious problem than with rats; the large ratio of surface area to volume of a mouse makes it particularly vulnerable to rapid heat loss.
Another consideration was whether a more natural escape strategy could be used, to facilitate learning. Since animals that fall into water and swim away from the safety of the shore are unlikely to pass on their genes, animals have evolved a natural tendency to swim to the edge of a body of water. The Morris water maze, however, requires them to swim to a hidden platform towards the center of the maze - exactly opposite to their evolved behavior. Therefore the paddling maze should incorporate escape to the edge of the apparatus. This feature, coupled with the use of relatively non-aversive shallow water, embodies the "Refinement" aspect of the "3 Rs" of Russell and Burch8.
Various types of maze design were tried; the common feature was that the water was always shallow (2 cm deep) and escape was via a tube piercing the transparent wall of the apparatus. Other tubes ("false exits") were also placed around the walls but these were blocked off. From the inside of the maze all false exits and the single true exit looked the same. Currently a dodecagonal (12-sided) maze is in use in Oxford, with 12 true/false exits set in the corners. In a recent development a transparent paddling Y-maze has been tested successfully.

Shallow Water Paddling Variants of Water Maze Tests in Mice

Mice can swim, but many strains appear to find this activity stressful. To overcome this problem mazes have been devised where escape from shallow water is used to motivate behaviour. These have been demonstrated to support learning at least as good as the traditional and widely used Morris water maze.

Farelerde Su Maze Testleri Sığ Su (kürek çekmeye) Türevleri

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

Davranış

Operant Procedures for Assessing Behavioral Flexibility in Rats

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these processes is the ability to adjust behavior in response to changes in one&#8217;s environment (i.e., behavioral flexibility). These processes are essential to normal human behavior, and may be disrupted in diverse neuropsychiatric conditions, including schizophrenia, alcoholism, depression, stroke, and Alzheimer&#8217;s disease. Understanding of the neurobiology of executive functions has been greatly advanced by the availability of animal tasks for assessing discrete components of behavioral flexibility, particularly strategy shifting and reversal learning. While several types of tasks have been developed, most are non-automated, labor intensive, and allow testing of only one animal at a time. The recent development of automated, operant-based tasks for assessing behavioral flexibility streamlines testing, standardizes stimulus presentation and data recording, and dramatically improves throughput. Here, we describe automated strategy shifting and reversal tasks, using operant chambers controlled by custom written software programs. Using these tasks, we have shown that the medial prefrontal cortex governs strategy shifting but not reversal learning in the rat, similar to the dissociation observed in humans. Moreover, animals with a neonatal hippocampal lesion, a neurodevelopmental model of schizophrenia, are selectively impaired on the strategy shifting task but not the reversal task. The strategy shifting task also allows the identification of separate types of performance errors, each of which is attributable to distinct neural substrates. The availability of these automated tasks, and the evidence supporting the dissociable contributions of separate prefrontal areas, makes them particularly well-suited assays for the investigation of basic neurobiological processes as well as drug discovery and screening in disease models.

The ability to assess executive functions such as behavioral flexibility in rats is useful for investigating the neurobiology of cognition in both intact animals and disease models. Here we describe automated tasks for assessing strategy shifting and reversal learning, which are particularly sensitive to disruptions in prefrontal cortical networks.

Sıçanlar Davranış Esneklik değerlendirilmesi için Edimsel Prosedürleri

Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these ...

The Forced Swim Test as a Model of Depressive-like Behavior

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like behavior in rodents. The FST is based on the assumption that when placing an animal in a container filled with water, it will first make efforts to escape but eventually will exhibit immobility that may be considered to reflect a measure of behavioral despair. This test has been extensively used because it involves the exposure of the animals to stress, which was shown to have a role in the tendency for major depression. Additionally, the FST has been shown to share some of the factors that are influenced or altered by depression in humans, including changes in food consumption, sleep abnormalities and drug-withdrawal-induced anhedonia. The main advantages of this procedure are that it is relatively easy to perform and that its results are easily and quickly analyzed. Moreover, its sensitivity to a broad range of antidepressant drugs that makes it a suitable screening test is one of the most important features leading to its high predictive validity. Despite its appeal, this model has a number of disadvantages. First, the issue of chronic augmentation is problematic in this test because in real life patients need to be treated for at least several weeks before they experience any relief from their symptoms. Last, due to the aversiveness of the FST, it is important to take into account possible influences it might have on brain structure/function if brain analyses are to be carried out following this procedure.

This protocol describes the forced swim test, which is used for the study of depressive-like behavior in rodents. This procedure involves placing an animal in a container filled with water that eventually will lead to the exhibition of immobility behavior, which is considered to reflect behavioral despair.

Depresif gibi Davranış Modeli olarak Zorla Swim Testi

The goal of the present protocol is to describe the forced swim test (FST), which is one of the most commonly used assays for the study of depressive-like ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

Çift H Maze: Kemirgenler A Sağlam Davranış Öğrenme için test ve Hafıza

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

A New Method for Inducing a Depression-Like Behavior in Rats

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is no existing format for studying the mechanism of action, prevention, containment, and treatment of contagious depression. The purpose of this study, therefore, was to establish the first animal model of contagious depression.
Healthy rats can contract depressive behaviors if exposed to depressed rats. Depression is induced in rats by subjecting them to several manipulations of chronic unpredictable stress (CUS) over 5 weeks, as described in the protocol. A successful sucrose preference test confirmed the development of depression in the rats. The CUS-exposed rats were then caged with na&#239;ve rats from the contagion group (1 na&#239;ve rat/2 depressed rats in a cage) for an additional 5 weeks. 30 social groups were created from the combination of CUS-exposed rats and na&#239;ve rats.
This proposed depression-contagion protocol in animals consists mainly of cohabiting CUS-exposed and healthy rats for 5 weeks. To ensure that this method works, a series of tests are carried out - first, the sucrose preference test upon inducing depression to rats, then, the sucrose preference test, alongside the open field and forced-swim tests at the end of the cohabitation period. Throughout the experiment, rats are given tags and are always returned to their cages after each test.
A few limitations to this method are the weak differences recorded between the experimental and control groups in the sucrose preference test and the irreversible traumatic outcome of the forced swim test. These may be worth considering for suitability before any future application of the protocol. Nonetheless, following the experiment, na&#239;ve rats developed contagion depression after 5 weeks of sharing the same cage with the CUS-exposed rats.

This protocol describes a new model by which healthy rats could contract depression over a given time periodthrough contagion by exposure to chronic unpredictable stressed (CUS) rats.

Bir depresyon gibi davranış Sıçanlarda Inducing için yeni bir yöntem

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is ...

Using Clicker Training and Social Observation to Teach Rats to Voluntarily Change Cages

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on well-being and can affect the comparability and reproducibility of research results, the amount of stress experienced by laboratory animals should be minimized and avoided when possible. Further, the direct contact between the rat and animal caretaker during the cage change bears hygiene risks and therefore possibly negatively impacts the well-being of the rats and the quality of the research. 
Our protocol aims to improve the routinely performed cage changing procedure. For this reason, we present a feasible protocol that enables rats to learn via clicker training and observation to voluntarily change to a clean cage. This training helps to reduce stress caused by the physical disturbance and handling associated with the cage changes and concurrently enables a reduction in direct contact between animal and animal caretaker after the training phase is completed.
The implementation of clicker training to rats is fast and easy. Rats are generally interested in the training and efficiently learn the desired behavior, which entails changing cages through a pipe. Even without training, the rats learn to perform the desired behavior by observation, as 80% of the observational learning group successfully changed cages when tested. The training further helps to establish a relationship of trust between trainer and animal. As hygiene and well-being are both very important in animal experiments, this protocol might also help to improve high-quality research.

This protocol introduces a method of cage change for rats via clicker training. Rats learn the desired behavior not only by direct training but also by observational learning. The implementation of this fast and easy protocol might help to improve well-being and hygiene in rodent facilities.

Tıkırtı eğitim ve sosyal gözlem gönüllü olarak kafes değiştirmek için fare öğretmek için kullanma

Cage cleaning is a routinely performed husbandry procedure and is known to induce stress in laboratory rats. As stress can have a negative impact on ...

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats

For many species, where status is a vital motivator that can affect health, social hierarchies influence behavior. Social hierarchies that include dominant-submissive relationships are common in both animal and human societies. These relationships can be affected by interactions with others and with their environment, making them difficult to analyze in a controlled study. Rather than a simple dominance hierarchy, this formation has a complicated presentation that allows rats to avoid aggression. Status can be stagnant or mutable, and results in complex societal stratifications. Here we describe a complex diving-for-food task to investigate rodent social hierarchy and behavioral interactions. This animal model may allow us to assess the relationship between a wide range of mental illnesses and social organization, as well as to study the effectiveness of therapy on social dysfunction.

This protocol describes a method of examining social hierarchy in a rat model. Rats perform a complex diving-for-food task in which they form a distinct hierarchy according to their willingness to dive underwater and swim to obtain a food pellet. This method is used to understand decision making and social relationships among highly social animals in small groups.

Sıçanlardaki Sosyal Organizasyonu ve Etkileşimleri Araştırmak İçin Karmaşık Bir Yemek İçin Dalış Görevi

For many species, where status is a vital motivator that can affect health, social hierarchies influence behavior. Social hierarchies that include ...

<meta charset="utf8"></head><body>Sıçan Yavrularının Refahı için Anne Egzersizini Keşfetmek

Swimming Exercise Protocol and Care Methods for Pregnant Rats

The developmental origins of health and disease concept highlights the impact of early environments on chronic non-communicable diseases like diabetes, cardiovascular disease, and cancer. Studies using animal models have investigated how maternal factors such as undernutrition, overnutrition, obesity, and exposure to chemicals or hypoxia affect fetal development and offspring health, leading to issues like low birth weight, high blood pressure, dyslipidemia, and insulin resistance. Given the increasing prevalence of overweight and obesity among reproductive-age women, effective interventions are critical. Maternal exercise during pregnancy has emerged as a key intervention, benefiting both mother and offspring and reducing the risk of disease. This study compares the differences of three exercise models on pregnant rats: voluntary wheel running, motorized treadmills, and swimming. Swimming is the most beneficial option due to its safe and controlled intensity levels. This protocol details the rat breeding methods, swimming training during pregnancy, and post-breeding nursing protocols. This model, suitable for various rat and mouse species, is useful for studying the benefits of maternal exercise on offspring health and intergenerational wellness.

<meta charset="utf8"></head><body>Yazar Spotlight: Hamilelik Sırasında Maternal Egzersiz ile Sıçanlarda Yavru Sağlığının Geliştirilmesi

Here, we present a protocol to delineate rat breeding methods, swimming training procedures, and post-breeding nursing protocols for pregnant rats after training. This protocol provides an animal model for studying the effects of maternal exercise during pregnancy on offspring and its underlying mechanisms.

Gebe Sıçanlarda Yüzme Egzersiz Protokolü ve Bakım Yöntemleri

The developmental origins of health and disease concept highlights the impact of early environments on chronic non-communicable diseases like diabetes, ...

Gönüllü Dalış Sualtı eğitim Rats: Memeli Dalış Tepki Araştırmalar

Özet

Daha Fazla Video Keşfet

Bu videodaki bölümler

Farelerde Su Maze Testleri Sığ Su (kürek çekmeye) Türevleri

Sıçanlar Davranış Esneklik değerlendirilmesi için Edimsel Prosedürleri

Depresif gibi Davranış Modeli olarak Zorla Swim Testi

Çift H Maze: Kemirgenler A Sağlam Davranış Öğrenme için test ve Hafıza

Bir depresyon gibi davranış Sıçanlarda Inducing için yeni bir yöntem

Tıkırtı eğitim ve sosyal gözlem gönüllü olarak kafes değiştirmek için fare öğretmek için kullanma

Sıçanlardaki Sosyal Organizasyonu ve Etkileşimleri Araştırmak İçin Karmaşık Bir Yemek İçin Dalış Görevi

Gebe Sıçanlarda Yüzme Egzersiz Protokolü ve Bakım Yöntemleri

Gebe Sıçanlarda Yüzme Egzersiz Protokolü ve Bakım Yöntemleri

Gebe Sıçanlarda Yüzme Egzersiz Protokolü ve Bakım Yöntemleri