Name: exposure of the pig cns for histological analysis: a manual for decapitation, skull opening, and brain removal
Uploaded: 2017-04-13T14:00:00
Description: Watch this Scientific Journal Video about Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal at JoVE.com

Die Autoren erkennen dankbar die geschickte Unterstützung von Frau Trine W. Mikkelsen, Frau Lise M. Fitting, und das Personal an Påskehøjgaard. Das dänische Medical Research Council, die Lundbeck-Stiftung, und die Novo Nordisk Foundation finanziell unterstützten die Studie.

Tier Anästhesie und euthanesia wurden gemäß „Principles of Labortierhaltung“ (NIH Publikation Nr 86-23, überarbeitet 1985) und genehmigt vom dänischen Rat für Tierforschung Ethik durchgeführt. 1. Instrumente <ol><li> Sammeln Sie die Instrumente in dem Video dargestellt und in der Tabelle der Materialien aufgeführt. </li></ol> 2. Enthauptung HINWEIS: Die Anästhesie wurde durch eine intramuskuläre Injektion von...

<ol>
	<li>Lind, M. N., Moustgaard, A., Jelsing, J., Vajta, G., Cumming, P., Hansen, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+pigs+in+neuroscience:+Modeling+brain+disorders.">The use of pigs in neuroscience: Modeling brain disorders.</a> Neurosci Biobehav Rev. 31, 728-751 (2007).</li><li>Bjarkam, C. R., 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=Neuromodulation+in+a+minipig+model+of+Parkinson+disease.">Neuromodulation in a minipig model of Parkinson disease.</a> British J Neurosurg. 22 (Suppl. 1), S9-S12 (2008).</li><li>Bjarkam, C. R., Cancian, G., Glud, A. N., Ettrup, K. S., &#216;stergaard, L., S&#248;rensen, J. 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R., 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=Safety+and+function+of+a+new+clinical+intracerebral+microinjection+instrument+for+stem+cells+and+therapeutics+examined+in+the+Göttingen+minipig.">Safety and function of a new clinical intracerebral microinjection instrument for stem cells and therapeutics examined in the Göttingen minipig.</a> Stereotact Funct Neurosurg. 88 (1), 56-63 (2010).</li><li>Fjord-Larsen, L., 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=Long-term+delivery+of+nerve+growth+factor+by+encapsulated+cell+biodelivery+in+the+minipig+basal+forebrain.">Long-term delivery of nerve growth factor by encapsulated cell biodelivery in the minipig basal forebrain.</a> Mol Therapy. 18 (12), 2164-2172 (2010).</li><li>S&#248;rensen, J. C., 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=Development+of+neuromodulation+treatments+in+a+large+animal+model+-+Do+neurosurgeons+dream+of+electric+pigs?.">Development of neuromodulation treatments in a large animal model - Do neurosurgeons dream of electric pigs?.</a> Prog Brain Res. 194, 97-103 (2011).</li><li>Dolezalova, D., 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=Pig+models+of+neurodegenerative+disorders:+utilization+in+cell+replacement-based+preclinical+safety+and+efficacy+studies.">Pig models of neurodegenerative disorders: utilization in cell replacement-based preclinical safety and efficacy studies.</a> J Comp Neurol. 522 (12), 2784-2801 (2014).</li><li>Mikkelsen, M., Moller, A., Jensen, L. H., Pedersen, A., Harajehi, J. 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H., Cumming, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pig+brain+stereotaxic+standard+space:+Mapping+of+cerebral+blood+flow+normative+values+and+effect+of+MPTP-lesioning.">Pig brain stereotaxic standard space: Mapping of cerebral blood flow normative values and effect of MPTP-lesioning.</a> Brain Res Bull. 66 (1), 17-29 (2005).</li><li>Glud, A. N., 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=Direct+gene+transfer+in+the+minipig+CNS+using+stereotaxic+lentiviral+microinjections.">Direct gene transfer in the minipig CNS using stereotaxic lentiviral microinjections.</a> Acta Neurobiol Exp. 70 (3), 1-8 (2010).</li><li>Glud, A. N., 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=Direct+MRI-guided+stereotaxic+viral+mediated+gene+transfer+of+alpha-synuclein+in+the+minipig+CNS.">Direct MRI-guided stereotaxic viral mediated gene transfer of alpha-synuclein in the minipig CNS.</a> Acta Neurobiol Exp. 71 (4), 508-518 (2011).</li><li>Ettrup, K. S., S&#248;rensen, J. C., Rodell, A., Alstrup, A. K. O., Bjarkam, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothalamic+deep+brain+stimulation+influences+autonomic+and+limbic+circuitry+involved+in+the+regulation+of+aggression+and+cardiocerebrovascular+control+in+the+minipig.">Hypothalamic deep brain stimulation influences autonomic and limbic circuitry involved in the regulation of aggression and cardiocerebrovascular control in the minipig.</a> Stereotact Funct Neurosurg. 90 (5), 281-291 (2012).</li><li>Nielsen, M. S., 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=Continuous+MPTP+intoxication+in+the+minipig+results+in+chronic+parkinsonian+deficits.">Continuous MPTP intoxication in the minipig results in chronic parkinsonian deficits.</a> Acta Neurobiol Exp. 76, 198-210 (2016).</li><li>Bjarkam, C. R., 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=A+MRI-compatible+stereotaxic+localizer+box+enables+high-precision+stereotaxic+procedures+in+pigs.">A MRI-compatible stereotaxic localizer box enables high-precision stereotaxic procedures in pigs.</a> J Neurosci Methods. 139 (2), 293-298 (2004).</li><li>Bjarkam, C. R., Jorgensen, R. L., Jensen, K. N., Sunde, N. A. A., S&#248;rensen, J. C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+brain+stimulation+electrode+anchoring+using+BioGlue&#174;,+a+protective+electrode+covering,+and+a+titanium+microplate.">Deep brain stimulation electrode anchoring using BioGlue&#174;, a protective electrode covering, and a titanium microplate.</a> J Neurosci Methods. 168, 151-155 (2008).</li><li>Ettrup, K. S., 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=Basic+Surgical+Techniques+in+the+Minipig:+Intubation,+Transurethral+Bladder+Catheterization,+Femoral+Vessel+Catheterization,+and+Transcardial+Perfusion.">Basic Surgical Techniques in the Minipig: Intubation, Transurethral Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion.</a> J Vis Exp. (52), e2652 (2011).</li><li>Ettrup, K. S., Torn&#248;e, J., S&#248;rensen, J. C., Bjarkam, C. 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H., 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=Positron+emission+tomography+of+living+brain+in+minipigs+and+domestic+pigs.">Positron emission tomography of living brain in minipigs and domestic pigs.</a> Scand J Lab Anim Sci Suppl. 25 (1), 127-135 (1998).</li><li>R&#248;hl, L., 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=Time+evolution+of+cerebral+perfusion+and+ADC+measured+by+MRI+in+a+porcine+stroke+model.">Time evolution of cerebral perfusion and ADC measured by MRI in a porcine stroke model.</a> J Magn Reson Imaging. 15 (2), 123-129 (2002).</li><li>Cumming, P., Gillings, N. M., Jensen, S. B., Bjarkam, C. 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Die meisten experimentellen Neurowissenschaften Studien werden in kleinen Tierarten, wie Mäuse und Ratten durchgeführt, in denen den Zugang zum ZNS durch einen dünnen Schädel- und Dural-Dicke erleichtert wird. In größeren Versuchstieren , wie Schweine 1, 4, 8, Schafe 32, aber, und nicht-menschliche Primaten, erfordern die beträchtlichen Dicke dieser Strukturen , die die Verwendung von robusten Ins...

<html> Translationale Neurowissenschaften Studien bei Schweinen immer beliebter in den letzten zwei Jahrzehnten worden. Die große Größe des Schweinehirns ermöglicht die Verwendung von herkömmlich klinischen brain Imager und der unmittelbaren Nutzung und Prüfung von neurochirurgischen Verfahren und Geräten aus der menschlichen Klinik 1, 2, 3, 4, 5, 6, 7, 8. In den letzten 20 Jahren, Schweine, insbesondere Minipigs (zB minipig Göttingen), wurden verwendet , neuromodulatorischen Behandlungsmodalitäten, wie beispielsweise Stammzelltransplantation zu untersuchen; viraler Vektor Transfektion; und tiefe Hirnstimulation gerichtet gegen Parkinson - Krankheit, Übergewicht, Depression und Alzheimer Krankheit 2, 6,= &quot;Xref&quot;&gt; 9, 10, 11, 12, 13, 14, 15, 16, 17. Dies wurde durch die Entwicklung von Stereotaxierahmen und chirurgische Ansätze folgte dem minipig CNS 3, 18, 19, 20, 21 zu manipulieren. Die eingeleiteten CNS Veränderungen in lebenden Tieren unter Verwendung von brain imaging (PET 10, 13, 22, 24 und MR 23), Zystometrie 11, 12, 25, Ganganalyse evaluiert17, neurologische Bewertung 9, 17 und Obduktion basierend auf der Histologie und stereologischen Analyse 14, 15, 17, 26, 27, 31. Jedoch erfordert postmortale Analyse die Belichtung und die Entfernung des Schweinehirns, das keine leichte Aufgabe ist, als ein dicker, knöcherner Schädel und ein faseriger dural das Schweinehirn abdeckt umgeben. Das Ziel dieses Papiers und Lehr-Video ist zu beschreiben, wie die Obduktion Schweinehirn und Hypophyse können unter Verwendung von nicht-motorisierte chirurgische Instrumente in 15-20 min in einem intakten Zustand belichtet und entfernt werden. Das Lehr-Video und fotografische Abbildungen zeigen männlich minipigs (Alter: 6 Monate, Körpergewicht: 20-25 kg) für eine anatomische Studie über die minipig Hypophyse verwendet.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heavy Scalpel Handle #4</td>
			<td style="width:191px;">FST (Fine Science Tools)</td>
			<td style="width:149px;">10008-13</td>
			<td style="width:481px;">Good for skin incision and soft tissue removal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Non-Sterile Scalpel Blades #23</td>
			<td style="width:191px;">FST&#160;</td>
			<td>10023-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scalpel Handle #7</td>
			<td style="width:191px;">FST&#160;</td>
			<td>10007-12</td>
			<td>Optimal for dural incision and precision work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Non-Sterile Scalpel Blades #11</td>
			<td style="width:191px;">FST&#160;</td>
			<td>10011-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgical Forceps</td>
			<td style="width:191px;">FST&#160;</td>
			<td>11024-18</td>
			<td>The tip of the surgical forceps ensure a firm grip&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Kerrison Bone Punch</td>
			<td style="width:191px;">Aesculap Neurosurgery</td>
			<td style="width:149px;">FF713R</td>
			<td>Must be robust, bite size 3-5 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bone Rongeur</td>
			<td>Aesculap Neurosurgery</td>
			<td style="width:149px;">MD615</td>
			<td>Must be robust, bite size 15 x 5 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bone Rongeur</td>
			<td>Aesculap Neurosurgery</td>
			<td style="width:149px;">FO551R</td>
			<td>Must be robust, bite size 25 x 15 mm&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bone Chisel</td>
			<td>Lawton</td>
			<td>67-0335</td>
			<td>The size of the chisel head should not exceed 20 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mallet (Hammer)</td>
			<td>Millarco</td>
			<td>5624108</td>
			<td>Weigth 300 g, length 30 cm, head hit area size 2 x 2 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro-Scissor</td>
			<td>FST&#160;</td>
			<td>14002-14&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissector</td>
			<td>Aesculap Neurosurgery</td>
			<td style="width:149px;">OL165R</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">&#160;G&#246;ttingen minipigs</td>
			<td style="width:191px;">&#160;Ellegaard G&#246;ttingen Minipigs A/S, Denmark</td>
			<td style="width:149px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Euthanimal</td>
			<td style="width:191px;"></td>
			<td style="width:149px;"></td>
			<td>pentobarbital</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine</td>
			<td style="width:191px;">Pfizer</td>
			<td style="width:149px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Midazolam&#160;</td>
			<td style="width:191px;">Hameln Pharmaceuticals</td>
			<td style="width:149px;"></td>
			<td></td>
		</tr>
	</tbody>
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exposure of the pig cns for histological analysis: a manual for decapitation, skull opening, and brain removal

Schweine haben in Großtier translationalen neurowissenschaftlichen Forschung als wirtschaftlich und ethisch möglich Ersatz für nichtmenschliche Primaten immer beliebter geworden. Die große Größe des Gehirn des Schweins ermöglicht die Verwendung von herkömmlich klinischem Gehirn Imager und der direkten Nutzung und Prüfung von neurochirurgischen Verfahren und Geräten aus der menschlichen Klinik. Weitere makroskopische und histologische Analyse erfordert jedoch postmortem Exposition des Schweins Zentralnervensystem (ZNS) und die anschließende Entfernung des Gehirns. Dies ist keine einfache Aufgabe, da das Schwein CNS durch einen dicken, knöchernen Schädels und der Wirbelsäule eingekapselt ist. Das Ziel dieses Papier und Lehr-Video ist zu beschreiben, wie die Obduktion Schweinehirn und die Hypophyse in einem intakten Zustand, geeignet für die anschließende makroskopische und histologische Analyse belichten und zu entfernen.

Um das Gewebematerial vor dem Austrocknen zu verhindern, wird empfohlen, das Gehirn und Hypophyse entfernt in einem Glas mit Fixativ oder isotonischer Kochsalzlösung gefüllt zu speichern, unmittelbar nach makroskopischer Analyse durchgeführt wurde. Das Gewebematerial kann in der Fixiermittel für Jahre gespeichert werden, während der Lagerung in isotonischer Kochsalzlösung, auch in einem Kühlschrank wird auf Gewebezerfall mit der Zeit führen. <p class="jove_content" fo:keep-to...

Watch this Scientific Journal Video about Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal at JoVE.com

Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal

Das Ziel dieses Papier und Lehr-Video ist zu beschreiben, wie die Obduktion Schweinehirn und Hypophyse in einem intakten Zustand, geeignet für die anschließende makroskopische und histologische Analyse belichten und zu entfernen.

Belichtung des Schweins CNS für die histologische Analyse: Ein Handbuch für Enthauptung, Schädel öffnen und Gehirn Removal

Pigs have become increasingly popular in large-animal translational neuroscience research as an economically and ethically feasible substitute to ...

Research

Education

JoVE Core

JoVE Journal

Anatomy and Physiology

Neuroscience

Unit 1

The Axial Skeleton

SCIENTISTS IN ACTION

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

Eine polierte und verstärkte ausgedünnten Schädel Fenster für Langzeit-Imaging im Gehirn der Maus

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form ...

Forschung

Neurowissenschaften

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

Co-Analyse von Hirnstruktur und-funktion mittels fMRT und Diffusionsgewichtete Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

A Comprehensive Protocol for Manual Segmentation of the Medial Temporal Lobe Structures

Eine umfassende Protokoll für die manuelle Segmentierung der medialen Temporallappen Structures

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, ...

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

Analyse Synaptische Modulation von<em&gt; Drosophila melanogaster</em&gt; Photorezeptoren nach der Einwirkung von Längerer Licht

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at ...

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

Hochdurchsatz-Analyse des Bewegungsapparates Verhaltens in der Drosophila -Insel-Assay

Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Aufnahme synaptische Plastizität in akuten Hippocampal Scheiben gepflegt in einem kleinvolumigen Recycling - Perfusions- und untertauchen-Typ-Kammer-System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...

Novel Passive Clearing Methods for the Rapid Production of Optical Transparency in Whole CNS Tissue

Neuartige passiv Clearing Methoden für die schnelle Produktion von optischen Transparenz im gesamten ZNS-Gewebe

Since the development of CLARITY, a bioelectrochemical clearing technique that allows for three-dimensional phenotype mapping within transparent tissues, ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Chronische Implantation des gesamten kortikalen Electrocorticographic Arrays in der Weißbüschelaffe

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Focused Ultrasound Induced Blood-Brain Barrier Opening for Targeting Brain Structures and Evaluating Chemogenetic Neuromodulation

Fokussierte ultraschallinduzierte Öffnung der Blut-Hirn-Schranke zur gezielten Behandlung von Hirnstrukturen und zur Beurteilung der chemogenetischen Neuromodulation

Acoustically Targeted Chemogenetics (ATAC) allows for the noninvasive control of specific neural circuits. ATAC achieves such control through a ...

Extraction and Dissection of the Domesticated Pig Brain

Extraktion und Zerlegung des domestizierenden Schweinehirns

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular ...