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

Forschung mit Mitteln des Midwestern University Office of Research und geförderten Programmen unterstützt. Auch dank der Midwestern University Tierhaltung und Erik Warren.

<ol>
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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>

Der Autor ist Berater für Stölting Company, und vorausgesetzt, das gesamte Design und die Spezifikationen der McCulloch Tauchflasche Maze, um sie für kommerzielle Zwecke.

Ratten in ihren verwilderten bilden können und tun, nutzen halbaquatischen Umwelt und oft tauchen unter Wasser, während der Nahrungssuche 6. So ist es nicht allzu überraschend, dass Ratten können sehr leicht trainiert, freiwillig unter Wasser tauchen werden. Die beschriebenen Trainingsverfahren kann bis 6 Wochen, die neu entwöhnt Ratten zu einer Körpergröße in den meisten erwachsenen Rattenhirn Atlanten verwendet (~ 300 g) bringen wird dauern. So sind die Gehirne von diesen ausgebildete Tiere werden leichter vergleichbar mit den in diesen Atlanten identifizierten anatomischen Strukturen sein. Nachdem sie im Startbereich platziert meisten Ratten werden ihre Unterwasser schwimmen innerhalb von 20 sec beginnen. Jedoch gelegentlich eine Ratte auf 5 min oder länger dauern bis vor Beginn ihres freiwilligen Tauchgang. Während es in diesem Stadium verlockend, die Ratten in den Unterwassertunnel zu erzwingen, dies in der Regel vermieden werden sollte, um die Ratten Zuordnen des Wasser mit einer negativen Erfahrungen zu verhindern. Nagetiere können stur sein und darf initiVerbündeter verweigern während der Trainingseinheiten zu tauchen, aber sobald sie erkennen, der einzige Weg, um das Wasser zu verlassen ist, indem Sie den Tauchgang durch das Labyrinth, sie in der Regel bald nach dem im Startbereich platziert initiieren ihre Tauchgänge. Ein kritischer Aspekt der Wiederholungen und methodischen Ausbildungsverfahren ist, dass sie in einer Weise, um die Spannung von den Ratten erlebten reduzieren geführt. Für Vermieter Ratten erkunden ihre Umwelt, vor allem während im Zielbereich zwischen Studien, scheint Stress weiter zu reduzieren. Ratten werden oft wieder in den Wasser zu schwimmen, und / oder untertauchen ihren Kopf unter Wasser noch auf dem Zielgelände Plattform sitzt. Dies legt nahe, dass Ratten sind nicht von Natur aus wasser aversiven. Zusätzlich ist es zwar nicht ungewöhnlich, Ratten, fäkale Pellets beim Schwimmen oder Tauchen oder beim Warten im Zielraum, tägliches Training führt zu weniger fäkale Pellets 17 zu produzieren. Im Allgemeinen ist die weniger gestresst die Ratten während der Ausbildung sind, desto weniger pellets sie zu produzieren. Etwaige fäkalen Pellets, die erzeugt werden, werden aus dem Wasser oder Zielbereich entfernt so bald wie möglich, um das Wasser relativ sauber zu halten. Ratten können gelegentlich zu blutigen Nasen beim Schwimmen und Tauchen, die ein Ergebnis der Insufflation von Wasser in die Nasengänge sein können. Das Auftreten von Blut kann aufgrund osmotischer Spannungen innerhalb der Nasenschleimhaut werden. Während im Zielbereich zwischen Studien die Ratten werden sich Bräutigam. Als Folge Blut aus der Nase kann über die Ratten Kopf und Schnauze umverteilt bekommen, was den Ratten eine leichte Rotstich, besonders um die Augen, im Laufe eines Trainings. Auch kann ein gelegentlicher Ratte Tauch stressig (auf dh durch Drehen um und verirren beim Tauchen unter Wasser (siehe Hinweis nach 5.4.3., Wie man dies zu) verhindern) zu finden und / oder negative Taucherlebnisse. In diesen Ratten können Porphyrin in den Ecken ihrer Augen erscheinen, signalisiert eine Stress response. Die Größe der Tauchtank wird zu einem gewissen Grad bestimmen die Raumanforderungen. Der beschriebene Behälter ist konstruiert für Ratten schwimmen 2-3 cm unter der Oberfläche des Wassers durch ein 5 m langes Labyrinth Plexiglas, eine Unterwasserschwimmdauer 10-15 sec 17,19,24,26,27 geben. Sollte ein Experiment entworfen werden, um Antworten von längerer Dauer Tauchgang zu messen, oder von einem tieferen Tauchwasser, muss der Tank erneut ausgeführt werden. Die Raumanforderungen könnten dann auch ändern, um die Dimensionen einer neu gestalteten Tauchtank passen. Wenn es keinen Bodenablauf in der Eingriffsraum zur Verfügung, kann das Wasser aus dem Tank in einem großen Behälter gesammelt werden, wie beispielsweise ein 60 gal Mülltonne, die dann an anderer Stelle in bequemer Weise entleert werden kann. Die Fos-Technik kann mit anderen neuronalen Nachweisverfahren verwendet werden, um weiter zu identifizieren und zu charakterisieren, die Teil Neuronen des Hirnstamms Schaltung des Sprungantwort. Beispielsweise,Und globosa Neuronen im lateralen A7 Bereich 26,27, die während freiwilliger aktiviert werden; Fos Erkennung in Verbindung mit Tyrosin-Hydroxylase-Färbung hat katecholaminerge Neuronen in der A1, C1, A2, A5 und Unter coeruleus Bereichen (26 Abbildung 6) identifiziert Tauchen. Auch hat Fos Erkennung in Verbindung mit der retrograden Tracers Cholera-Toxin die Zellkörper der Motoneuronen kardialen vagalen innerhalb der externen Bildung des Nucleus ambiguus, die bei freiwilligen Tauch 20 aktiviert werden, identifiziert. Untersuchung des zentralen Nerven Integration der kardiorespiratorischen antwortet Tauch ist für eine Reihe von Gründen, 6,8,28. Die Tauchantwort ermöglicht Tieren, einschließlich des Menschen, zu untergetaucht Unterwasser ohne Atem für längere Zeit bleiben. Die Tauchantwort stellt eine funktionelle Reorganisation der Hirnstamm homöostatischen Kontrolle, und zeigt eine der mächtigsten paMuste r der bei Tieren beobachteten vegetativen Reflexe. Das Tauchen Antwort kann auch wichtige klinisch beim Menschen als Teil der trigemino-kardialen Reflex, Nasen-Rachen-Reflex, und / oder plötzlichen Kindstod sein. Schließlich wird ein Verständnis der neuronalen Schaltkreise, die im Hirnstamm von Ratten existiert helfen festzustellen, wie kortikalen Afferenzen können grundlegende Hirnstamm vegetativen Reflexe modifizieren. All diese Überlegungen machen Studie der zentralen Aspekte des Säugetiertauchantwort inhärent sinnvoll und interessant. Mit den beschriebenen Verfahren an Ratten zu trainieren, um sich freiwillig zu tauchen unter Wasser besser Untersuchung der zentralen Aspekte des Säugetiertauchantwort als die Verwendung von Zwangs tauchte Tieren ermöglichen. Denn die Trainingsverfahren wie beschrieben 1) reduzieren Aktivierung von ZNS Stress Schaltung und 2) haben CNS nicht aktiviert Lohn-Schaltung, da externe Belohnungen werden nicht verwendet.

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.

Erfolgreicher Abschluss der beschriebenen Schwimmen und Tauchen Trainingsverfahren können den Stress von Ratten erlebt beim Tauchen unter Wasser zu verringern. Blut Corticosteron Ebenen zeigen, dass wiederholte tägliche Training verringert den Leidhaftigkeit mit freiwilligen Tauchen verbunden sind, und ausgebildete Ratten finden Tauchen nicht mehr Stress als die täglich von einem Menschen (Abbildung 2; 17) abgewickelt. Umgekehrt, Ratten nicht in der Tauchprotokoll geschult finden freiwillige Tauch stressig (Abbildung 2; 17). Zusätzlich beide trainierten und untrainierten Ratten finden gezwungen Tauchen bis der größte Stressfaktor (Abbildung 2; 17) sein. Und Hinter Kanülen (Abbildung 4; 21-23), die Herz-Kreislauf Antworten von Schwimmen, Tauchen und freiwillige Zwangstauch Ratten wurden mit implantierten Telemetriegeräte (8,17-20 Abbildung 3) aufgezeichnet. Unmittelbar nach freiwilligenUntertauchen, und innerhalb eines einzigen Schlag, Herzfrequenz sinkt um 78% und der mittlere arterielle Blutdruck sinkt um 25% 17. Diese Ergebnisse zeigen, dass freiwillig tauchen Ratten zeigen die gleichen kardiorespiratorische Veränderungen typischerweise in anderen Tauch Tieren gesehen. Hinter arteriellen Kanülen wurden verwendet, um die Muscarin-Antagonist Atropin, der die Bradykardie mit freiwilligen Tauchen (4; 21) zugeordnet eliminiert zu injizieren, und um die Verteilung der Herzleistung 22 einschließlich des zerebralen Blutflusses 23, bei freiwilligen Tauch bestimmen. Hinter Kanülen wurden auch verwendet, um zu zeigen, dass Ratten ignorieren zunehmende arterielle Hypoxämie und Hyperkapnie während sie untergetaucht sind 18, und das bereits bestehende Chemorezeptor-Laufwerk hat keine Auswirkungen auf das kardiovaskuläre Reaktionen auf freiwilliger Tauch 21 haben. Neuronen innerhalb Laminae I und II der ventralen Markhinterhorn (MDH) auszudrücken Fos during freiwillige Tauchen und diese Neuronen kann die anfängliche Hirnstamm afferenten Relais des Tauchantwort (; 24 Abbildung 5) bilden. Wichtige Hirnstamm kardiorespiratorischen Kontrollbereiche, wie der Schwanzkompressorbereich (CPA), Nucleus tractus Einsiedler (NTS), rostral ventrolateralen Medulla (RVLM) und peribrachial Regionen erhöht alle anzeigen Fos Kennzeichnung während freiwillige Tauch verglichen mit Schwimm 25. Neuronen in chemosensitiven Regionen des Hirnstamms auszudrücken Fos nach langer Dauer gezwungen Tauchgänge 18. <img alt="Figur 1" fo:content-width="4in" src="/files/ftp_upload/52093/52093fig1highres.jpg" width="400" /> Abb. 1: Schematische Darstellung der Tauchflasche Eine Plexiglastank (100 x 60 x 15 cm) wurde verwendet, um ein einfaches Labyrinth, bestehend aus fünf 1 m lange Kanäle zu erstellen. Der Tank wurde mit 30 ° C Leitungswasser gefüllt, und die Ratten wurden zunächst negotiat ausgebildetene das Labyrinth durch Schwimmen auf der Oberfläche des Wassers, im Start-Bereich (oben links) mit dem Zielbereich (unten rechts). Die Ratten wurden dann geschult, um durch das Labyrinth zu tauchen, hielt unter Wasser durch horizontale Plexiglas Stücke platziert 2-3 cm unter der Wasseroberfläche. [Diese Zahl hat sich von 26 modifiziert worden] <img alt="Figur 2" fo:content-width="5in" src="/files/ftp_upload/52093/52093fig2highres.jpg" width="500" /> Figur 2: Corticosteron-Messungen. Blutentnahmen aus Rattenschwanzvenen wurden verwendet, um Corticosteron-Konzentrationen (Mittelwert ± SE) von Ratten in ihren Käfigen (Naive) links messen, Ratten für 10 min / Tag (behandelt werden), Ratten darauf trainiert, schwimmen und tauchen (Ausgebildete) und Ratten behandelt dass erhielt keine schwimmen oder tauchen Anlernen (ungelernter). Corticosteron wurde gemessen, nachdem ausgebildete Ratten hatten ihre Schwimmtraining (Links bar eingestellt) abgeschlossen hatte, nachdem ausgebildete Ratten ihre freiwillige div abgeschlossene Ausbildung (Zentrum Reihe von Balken), und nach trainiert Ratten hatten ihre Zwangstauchausbildung (Recht bar eingestellt) abgeschlossen. 1 zeigt Wert ist deutlich größer als naiv; 2 zeigt Wert ist deutlich größer als behandeltes; 3 zeigt Wert ist deutlich größer als Ausgebildete; * Bedeutet, dass bei trainierten Ratten Wert bei Zwangs Tauchgang ist deutlich größer als bei der freiwilligen Tauchgang. [Diese Zahl hat sich von 17 modifiziert worden] <img alt="Figur 3" fo:content-width="5in" src="/files/ftp_upload/52093/52093fig3highres.jpg" width="500" /> Abbildung 3: Der arterielle Blutdruck Spuren von telemetrischen Sendern Rohdaten Spuren, pulsierenden arteriellen Blutdruck während Schwimmen (linke Spalte), freiwillige Tauchen (mittlere Spalte) und zwang Tauchen (rechte Spalte) von Ratten darauf trainiert, schwimmen und tauchen durch das Labyrinth. (untere Reihe) und von Ratten, die nicht die trai hattening-Verfahren (obere Reihe). Tauchen unter Wasser (sowohl freiwillige und erzwungene Untertauchen) produziert eine sofortige Bradykardie und langsameren Beginn Anstieg des arteriellen Drucks, während Schwimmen an der Oberfläche des Wassers verursacht keine solchen kardiovaskulären Veränderungen. Bars unter Spuren zeigen Perioden Flutung. Breaks in Spuren zeigen, die Zeiträume, telemetrische Signal verloren ging. [Diese Zahl hat sich von 17 modifiziert worden] <img alt="Figur 4" fo:content-width="5in" src="/files/ftp_upload/52093/52093fig4highres.jpg" width="500" /> Abbildung 4: Atropin beseitigt Tauch Bradykardie. Originalaufnahmen der pulsierenden arteriellen Blutdruck von freiwillig tauchen Ratten (A) vor und (B) nach Atropin-Vorbehandlung. Spuren wurden unter Verwendung eines Hinterarterienkanüle erhalten. Bevor Atropin Vorbehandlung, verminderte arterielle Druck etwas nach Untertauchen, aber tHenne erhöht, um größer als vor dem Tauchgang für den Rest des Tauchgangs. Die Herzfrequenz wurde aus benachbarten Pulsdruck Abständen bestimmt. Beim Untertauchen gab es eine unmittelbare und wesentliche Bradykardie, die für die Dauer des Tauchgangs aufrechterhalten wurde. Nach parasympathische Blockade durch Atropin Vorbehandlung der Bradykardie wurde eliminiert. Es gab auch eine Erhöhung des arteriellen Drucks während des Tauchgangs. Die Bar unter der Messkurve stellt den Zeitraum von Flutung. [McCulloch, unveröffentlicht] <img alt="Figur 5" fo:content-width="6in" src="/files/ftp_upload/52093/52093fig5highres.jpg" width="600" /> Abbildung 5: Fos Etikettierung im MDH. Mikrofotografien des Trigeminus Markhinterhorn (MDH) und dem Rücken Trigeminus-Trakt (SP5) bei Ratten darauf trainiert, tauchen unter Wasser. (A) In einer Kontrollratte, die nicht wiederholt tauchen habe es keine Fos Etikettierung. (B) in einem Schwimmratte ist sehr beleuchtettle Fos Etikett in der MDH (große Pfeilspitze) oder paratrigeminal Kern (kleiner Pfeil) in sp5. (C) In einem Tauchratten es mehr Fos Kennzeichnung ventral sowohl in der MDH (große Pfeilspitze) und paratrigeminal Kern (kleine Pfeile) im Vergleich zu das Schwimmen und Kontrollratte. Legen Sie im Panel (A) zeigt die rostral-kaudale Lage des Panels AC. Maßstabsbalken in Panel C ist 100 um. [Diese Zahl hat sich von 24 modifiziert worden] <img alt="Figur 6" fo:content-width="5in" src="/files/ftp_upload/52093/52093fig6highres.jpg" width="500" /> Abbildung 6: Aktiviert katecholaminerge Neuronen vom Tauchen Ratten. Mikrofotografien zeigen die medullas eines nichttauchKontrollRatte (A, C und E) und einer freiwillig Tauch Ratte (B, D und F). Das Hirngewebe wurde immunhistologisch sowohl eine Fos verarbeitetd Tyrosinhydroxylase (TH), Herstellung braun und schwarz TH somas Fos Kernen. Offene Pfeilspitzen identifizieren einfach markierten TH-positiven Neuronen, während durchgezogene Pfeile identifizieren Fos + TH doppelt markierten Neuronen. A1 Neuronen sind in A und B. C1 Neuronen identifiziert werden in C identifiziert und D. A5 Neuronen in E und F. Weitere Fos und TH doppelt markierten sind in der A1, C1 gesehen identifiziert und A5 Regionen der Tauch Ratte als in der nichttauchKontrollRatte. Kalibrierung Bar in E ist für Platten AF und 250 &amp; mgr; m. Kalibrierung Bar im Einschub in F ist für alle Einsätze und 50 um. [Diese Zahl hat sich von 26 modifiziert worden]

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.

Trainings Ratten Freiwillig Dive Unterwasser: Untersuchungen des Mammalian Diving Antwort

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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12.2K Aufrufe.  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.

Serie: 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.

Shallow Water (Paddeln) Varianten of Water Maze-Tests in Mäuse

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 ...

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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.

Operante Verfahren zur Bewertung der Verhaltensflexibilität bei Ratten

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.

Forced Swim-Test als ein Modell der Depressive ähnliche Verhalten

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.

Die Doppel-H Maze: A Robust Verhaltenstest für Lernen und Gedächtnis in Nagetiere

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.

Eine neue Methode zur Induktion einer Depression-ähnliches Verhaltens bei Ratten

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.

Verwendung von Clicker-Training und soziale Beobachtung beibringen Ratten Käfige freiwillig ändern

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.

Eine komplexe Tauch-für-Lebensmittel-Aufgabe zur Untersuchung der sozialen Organisation und Interaktionen bei Ratten

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>Erforschung der Bewegung von Müttern für das Wohlbefinden des Rattennachwuchses

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>Autor im Rampenlicht: Verbesserung der Gesundheit der Nachkommen bei Ratten durch Bewegung der Mutter während der Schwangerschaft

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.

Schwimmübungsprotokoll und Pflegemethoden für trächtige Ratten

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

Trainings Ratten Freiwillig Dive Unterwasser: Untersuchungen des Mammalian Diving Antwort

Zusammenfassung

Weitere Videos entdecken

Kapitel in diesem Video

Shallow Water (Paddeln) Varianten of Water Maze-Tests in Mäuse

Operante Verfahren zur Bewertung der Verhaltensflexibilität bei Ratten

Forced Swim-Test als ein Modell der Depressive ähnliche Verhalten

Die Doppel-H Maze: A Robust Verhaltenstest für Lernen und Gedächtnis in Nagetiere

Eine neue Methode zur Induktion einer Depression-ähnliches Verhaltens bei Ratten

Verwendung von Clicker-Training und soziale Beobachtung beibringen Ratten Käfige freiwillig ändern

Eine komplexe Tauch-für-Lebensmittel-Aufgabe zur Untersuchung der sozialen Organisation und Interaktionen bei Ratten

Schwimmübungsprotokoll und Pflegemethoden für trächtige Ratten

Schwimmübungsprotokoll und Pflegemethoden für trächtige Ratten

Schwimmübungsprotokoll und Pflegemethoden für trächtige Ratten