Name: retrograde neuroanatomical tracing of phrenic motor neurons in mice
Uploaded: 2018-02-22T14:00:00
Description: Watch this Scientific Journal Video about Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice at JoVE.com

Vi er taknemmelige for Robert Graffin og Pauline Duhant deres tekniske support.

En eksperimentel protokol blev gennemført i overensstemmelse med de Europæiske Fællesskabers Rådets direktiver for dyr eksperiment (2010/63/EU, 86/609/EOEF og 87-848/EØF) og blev godkendt af den dyr etiske udvalg af Universitet Namur (etik projekt n ° 17-284 ). Figur 1 viser de to respektive tilgange: transdiaphragmatic eller transthoracic injektioner. Brug mandlige C57bl/6J mus (n = 18), i alderen fra 3 til 4 måneder i studiet.
1. forberedelse af CTB løsnin...

<ol>
	<li>Webber, C. L., Wurster, R. D., Chung, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cat+phrenic+nucleus+architecture+as+revealed+by+horseradish+peroxidase+mapping.">Cat phrenic nucleus architecture as revealed by horseradish peroxidase mapping.</a> Exp Brain Res. 35 (3), 395-406 (1979).</li><li>Goshgarian, H. G., Rafols, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+phrenic+nucleus+of+the+albino+rat:+a+correlative+HRP+and+Golgi+study.">The phrenic nucleus of the albino rat: a correlative HRP and Golgi study.</a> J Comp Neurol. 201 (3), 441-456 (1981).</li><li>Gordon, D. C., Richmond, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topography+in+the+phrenic+motoneuron+nucleus+demonstrated+by+retrograde+multiple-labelling+techniques.">Topography in the phrenic motoneuron nucleus demonstrated by retrograde multiple-labelling techniques.</a> J Comp Neurol. 292 (3), 424-434 (1990).</li><li>Mantilla, C. B., Zhan, W. Z., Sieck, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retrograde+labeling+of+phrenic+motoneurons+by+intrapleural+injection.">Retrograde labeling of phrenic motoneurons by intrapleural injection.</a> J Neurosci Methods. 182 (2), 244-249 (2009).</li><li>Nicaise, 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=Early+phrenic+motor+neuron+loss+and+transient+respiratory+abnormalities+after+unilateral+cervical+spinal+cord+contusion.">Early phrenic motor neuron loss and transient respiratory abnormalities after unilateral cervical spinal cord contusion.</a> J Neurotrauma. 30 (12), 1092-1099 (2013).</li><li>Nicaise, 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=Phrenic+motor+neuron+degeneration+compromises+phrenic+axonal+circuitry+and+diaphragm+activity+in+a+unilateral+cervical+contusion+model+of+spinal+cord+injury.">Phrenic motor neuron degeneration compromises phrenic axonal circuitry and diaphragm activity in a unilateral cervical contusion model of spinal cord injury.</a> Exp Neurol. 235 (2), 539-552 (2012).</li><li>Nicaise, 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=Degeneration+of+phrenic+motor+neurons+induces+long-term+diaphragm+deficits+following+mid-cervical+spinal+contusion+in+mice.">Degeneration of phrenic motor neurons induces long-term diaphragm deficits following mid-cervical spinal contusion in mice.</a> J Neurotrauma. 29 (18), 2748-2760 (2012).</li><li>Qiu, K., Lane, M. A., Lee, K. Z., Reier, P. J., Fuller, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+phrenic+motor+nucleus+in+the+adult+mouse.">The phrenic motor nucleus in the adult mouse.</a> Exp Neurol. 226 (1), 254-258 (2010).</li><li>Laskowski, M. B., Sanes, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topographic+mapping+of+motor+pools+onto+skeletal+muscles.">Topographic mapping of motor pools onto skeletal muscles.</a> J Neurosci. 7 (1), 252-260 (1987).</li><li>Feldman, J. L., Loewy, A. D., Speck, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Projections+from+the+ventral+respiratory+group+to+phrenic+and+intercostal+motoneurons+in+cat:+an+autoradiographic+study.">Projections from the ventral respiratory group to phrenic and intercostal motoneurons in cat: an autoradiographic study.</a> J Neurosci. 5 (8), 1993-2000 (1985).</li><li>Gottschall, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+diaphragm+of+the+rat+and+its+innervation.+Muscle+fiber+composition;+perikarya+and+axons+of+efferent+and+afferent+neurons.">The diaphragm of the rat and its innervation. Muscle fiber composition; perikarya and axons of efferent and afferent neurons.</a> Anat Embryol (Berl). 161 (4), 405-417 (1981).</li><li>Lai, B. Q., 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=Cholera+Toxin+B+Subunit+Shows+Transneuronal+Tracing+after+Injection+in+an+Injured+Sciatic+Nerve.">Cholera Toxin B Subunit Shows Transneuronal Tracing after Injection in an Injured Sciatic Nerve.</a> PLoS One. 10 (12), e0144030 (2015).</li><li>Torgersen, M. L., Skretting, G., van Deurs, B., Sandvig, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Internalization+of+cholera+toxin+by+different+endocytic+mechanisms.">Internalization of cholera toxin by different endocytic mechanisms.</a> J Cell Sci. 114 (Pt 20), 3737-3747 (2001).</li><li>Chinnapen, D. J., Chinnapen, H., Saslowsky, D., Lencer, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rafting+with+cholera+toxin:+endocytosis+and+trafficking+from+plasma+membrane+to+ER.">Rafting with cholera toxin: endocytosis and trafficking from plasma membrane to ER.</a> FEMS Microbiol Lett. 266 (2), 129-137 (2007).</li><li>Fujinaga, Y., 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=Gangliosides+that+associate+with+lipid+rafts+mediate+transport+of+cholera+and+related+toxins+from+the+plasma+membrane+to+endoplasmic+reticulm.">Gangliosides that associate with lipid rafts mediate transport of cholera and related toxins from the plasma membrane to endoplasmic reticulm.</a> Mol Biol Cell. 14 (12), 4783-4793 (2003).</li><li>Badizadegan, K., Wheeler, H. E., Fujinaga, Y., Lencer, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trafficking+of+cholera+toxin-ganglioside+GM1+complex+into+Golgi+and+induction+of+toxicity+depend+on+actin+cytoskeleton.">Trafficking of cholera toxin-ganglioside GM1 complex into Golgi and induction of toxicity depend on actin cytoskeleton.</a> Am J Physiol Cell Physiol. 287 (5), C1453-C1462 (2004).</li><li>Abbott, C. J., 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=Imaging+axonal+transport+in+the+rat+visual+pathway.">Imaging axonal transport in the rat visual pathway.</a> Biomed Opt Express. 4 (2), 364-386 (2013).</li><li>Li, K., 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=Overexpression+of+the+astrocyte+glutamate+transporter+GLT1+exacerbates+phrenic+motor+neuron+degeneration,+diaphragm+compromise,+and+forelimb+motor+dysfunction+following+cervical+contusion+spinal+cord+injury.">Overexpression of the astrocyte glutamate transporter GLT1 exacerbates phrenic motor neuron degeneration, diaphragm compromise, and forelimb motor dysfunction following cervical contusion spinal cord injury.</a> J Neurosci. 34 (22), 7622-7638 (2014).</li><li>Lepore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraspinal+cell+transplantation+for+targeting+cervical+ventral+horn+in+amyotrophic+lateral+sclerosis+and+traumatic+spinal+cord+injury.">Intraspinal cell transplantation for targeting cervical ventral horn in amyotrophic lateral sclerosis and traumatic spinal cord injury.</a> J Vis Exp. (55), (2011).</li><li>Llado, J., 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=Degeneration+of+respiratory+motor+neurons+in+the+SOD1+G93A+transgenic+rat+model+of+ALS.">Degeneration of respiratory motor neurons in the SOD1 G93A transgenic rat model of ALS.</a> Neurobiol Dis. 21 (1), 110-118 (2006).</li><li>Lepore, A. 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=Focal+transplantation-based+astrocyte+replacement+is+neuroprotective+in+a+model+of+motor+neuron+disease.">Focal transplantation-based astrocyte replacement is neuroprotective in a model of motor neuron disease.</a> Nat Neurosci. 11 (11), 1294-1301 (2008).</li><li>Sieck, G. C., Fournier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diaphragm+motor+unit+recruitment+during+ventilatory+and+nonventilatory+behaviors.">Diaphragm motor unit recruitment during ventilatory and nonventilatory behaviors.</a> J Appl Physiol. 66 (6), 2539-2545 (1989).</li><li>Janicot, M., Clot, J. P., Desbuquois, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+of+cholera+toxin+with+isolated+hepatocytes.+Effects+of+low+pH,+chloroquine+and+monensin+on+toxin+internalization,+processing+and+action.">Interactions of cholera toxin with isolated hepatocytes. Effects of low pH, chloroquine and monensin on toxin internalization, processing and action.</a> Biochem J. 253 (3), 735-743 (1988).</li></ol>

Protokollen beskrevet heri kan anvendes til en stamme af voksen mus eller enhver eksperimentelle paradigme integriteten af membran-PhMN kredsløb bør evalueres. For eksempel, er Amyotrofisk lateral sklerose (ALS) og cervikal rygmarvsskade (cSCI) betingelser forbundet med PhMN tab, anterograd degeneration af phrenic axoner og efterfølgende respiratorisk kompromis. Dyremodeller for ALS eller cSCI efterligne histopatologiske og funktionelle respiratorisk underskud observeret i menneskelige sygdomme. I disse modeller, der ...

Formålet med undersøgelsen er at præsentere en pålidelig metode til at identificere phrenic motor neuroner (PhMN) på mus rygmarven sektioner. Injektion af en fluorescerende neuroanatomical sporstof i pleural hulrum blev valgt som leveringsmetode til at nå de phrenic neuromuskulære fremskrivninger på mellemgulvet og bruge retrograd transport langs de phrenic axoner til at mærke phrenic celle organer. To teknikker til intrapleural levering er beskrevet: transdiaphragmatic versus transthoracic.
Phrenic motoriske neuroner er spinal relæ celler hvis axoner konvergerer i phrenic nerver, som i sidste ende innerverer mellemgulvet. Disse er lavere motor neuroner modtager inspiratory drevet fra de bulbar respiratoriske Centre og formidle det til mellemgulvet neuro-muskulære vejkryds (NMJ). PhMN er opbygget i to motor kolonner, en for hver hemicord, løber langs de midten af halshvirvelsøjlen. I de fleste pattedyr, herunder mennesker, spredes de phrenic motor kolonner fra niveauer C3 til C61,2,3. Vi og andre har bekræftet, at PhMN koncentreret i C3-C5 niveauer i rotter og mus rygmarven4,5,6,7,8. Den topografiske fordeling af phrenic celler er ikke tilfældigt; motoriske neuroner innerverer den brystbenet del af mellemgulvet er fordelt mere tæt i den kranielle del af phrenic motor pool (C3), motoriske neuroner innerverer den crural del er mere caudale (C5)9. PhMN er endvidere grupperet skiftevis i ventrale horn grå materie. På C3 plan, klyngerne af phrenic celler ligger lateralt, så de skift i retningen til ventrolaterale og findes ventromedially på de mest caudale niveauer10,11.
Givet deres afgørende rolle under inspiration, er det af yderste vigtighed at identificere PhMN i sund rygmarven men også følge deres skæbne under patologiske tilstande, som degenerative sygdomme eller traumatiske skader på rygmarven. Da PhMN ikke afviger morfologisk fra andre cervikal motoriske neuroner, identifikation af PhMN er afhængig af målrettet levering af neuroanatomical røbestoffer enten på niveauet af primære respiratoriske Centre8, eller på mellemgulvet NMJ7 eller i phrenic nerve4. Sporstof er taget op af nervefibre og transporteres op til de phrenic celle organer i halshvirvelsøjlen, hvor det kan visualiseres ved hjælp af direkte eller indirekte detektionssystemer. Retrograd eller anterograd røbestoffer er kommercielt tilgængelige med en bred vifte af konjugater. Bemærkelsesværdigt, hver tracer er begavet med nej, lav eller høj evner for trans synaptic sporing.
I den aktuelle undersøgelse valgte vi beta underenheden af kolera toksin (CTB) functionalized med Alexa Fluor 555 (herefter benævnt CTB-fluorophore) som en fluorescerende etiket, giver mulighed for en direkte visualisering af PhMN på frosne rygmarven sektioner. CTB er som regel beskrevet som en monosynaptic tracer selvom forsøgsdata tendens til at vise en transneuronal passage12. CTB har evnen til at binde ganglioside GM1 på plasma membran af nerve ender. CTB er internaliseret via clathrin-afhængige eller -uafhængige mekanismer og trafikker gennem trans-Golgi netværk i det endoplasmatiske reticulum i en retrograd mode13,14. Internalisering og retrograd transport synes at være afhænger actin cytoskeleton15,16 samt mikrotubulus netværk17.
For at påvise nytten af CTB som en retrograd neuroanatomical tracer mærkning membran-PhMN kredsløb, blev CTB-fluorophore leveret intrapleurally. CTB blev administreret ved hjælp af to metoder: den ene omfattede en laparotomi og flere transdiaphragmatic injektioner; den anden, mindre invasive, brugte en unik transthoracic injektion. Fire dage senere, fluorescently-mærket PhMNs var kvantificeres i de cervikale rygmarv fra både fra raske og spinally sårede (C4) dyr.

<table>
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		<tr>
			<td>Glass-bead sterilizer Steri 250</td>
			<td>Keller</td>
			<td>31-101</td>
			<td></td>
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		<tr>
			<td>Small scissors</td>
			<td>F.S.T.</td>
			<td>14058-00</td>
			<td></td>
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			<td>Soft tweezers</td>
			<td>F.S.T.</td>
			<td>11042-08</td>
			<td></td>
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			<td>Scalpel blades</td>
			<td>Swann Morton</td>
			<td>No.11 or 15</td>
			<td></td>
		</tr>
		<tr>
			<td>Cholera toxin subunit beta conjugated to Alexa Fluor 555</td>
			<td>Life Technologies</td>
			<td>C22843</td>
			<td>Bring at room temperature before use&#160;</td>
		</tr>
		<tr>
			<td>10ul Hamilton syringue, removable needle</td>
			<td>Sigma-Aldrich</td>
			<td>701RN</td>
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			<td>33-gauge needle for Hamilton syringue, 20mm length, point style 4</td>
			<td>Filter Service</td>
			<td>7803-05</td>
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			<td>500ul insulin syringue MyJector, 27-gauge</td>
			<td>Terumo</td>
			<td>BS05M2713</td>
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			<td>V.W.R.</td>
			<td>631-0995</td>
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			<td>Resorbable 4/0 sutures</td>
			<td>S.M.I. AG</td>
			<td>15151519</td>
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			<td>Needle holder</td>
			<td>F.S.T.</td>
			<td>12002-14</td>
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			<td>9mm autoclips</td>
			<td>Bioseb</td>
			<td>205016</td>
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			<td>Autoclip 9mm applier</td>
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			<td>MikRon 9mm</td>
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</table>

retrograde neuroanatomical tracing of phrenic motor neurons in mice

Phrenic motoriske neuroner er cervikal motoriske neuroner stammer fra C3 til C6 niveauer i mest pattedyrarter. Cytoskeletale fremskrivninger konvergerer i phrenic nerver, der innerverer de respiratoriske membran. I rygmarven skiver, ikke kan phrenic motoriske neuroner identificeres fra andre motoriske neuroner på morfologiske eller biokemiske kriterier. Vi leverer beskrivelse af procedurer til at visualisere phrenic motorneuron celle organer i mus, følgende intrapleural injektioner af kolera toksin subunit beta (CTB) konjugeret med et fluorophore. Denne fluorescerende neuroanatomical tracer har mulighed for at blive fanget på mellemgulvet neuromuskulære junction, udføres retrogradely langs de phrenic axoner og nå de phrenic celle organer. To metodiske tilgange til intrapleural CTB levering sammenlignes: transdiaphragmatic versus transthoracic injektioner. Begge tilgange er vellykket og resultere i tilsvarende antal CTB-mærket phrenic motor neuroner. Afslutningsvis kan disse teknikker anvendes for at visualisere eller kvantificere phrenic motoriske neuroner i forskellige eksperimentelle undersøgelser som dem, der fokuserede på den membran-phrenic kredsløb.

Mandlige C57bl/6J mus (n = 18), i alderen fra 3 til 4 måneder blev inkluderet i undersøgelsen. På dag 0 af eksperimentet undergik 8 mus en ensidig C4 kontusion, højresidig, ifølge offentliggjorte protokol7,18. Som fingeret procedure undergik 10 mus en laminektomi på toppen af C4 uden kontusion. På dag 3, blev mus forberedt til intrapleural injektioner af CTB-fluorophore efter to forskellige procedurer beskrevet ovenfor. På...

Watch this Scientific Journal Video about Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice at JoVE.com

Retrograde Neuroanatomical Tracing of Phrenic Motor Neurons in Mice

Her, beskriver vi en protokol til at identificere phrenic motoriske neuroner i mus efter intrapleural levering af fluorophore konjugeret kolera toksin subunit beta. To teknikker er sammenlignet for at injicere den pleural hulrum: transdiaphragmatic versus transthoracic tilgange.

Retrograd Neuroanatomical sporing af Phrenic motoriske neuroner i mus

Phrenic motor neurons are cervical motor neurons originating from C3 to C6 levels in most mammalian species. Axonal projections converge into phrenic ...

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