Name: quantifying acute changes in renal sympathetic nerve activity in response to central nervous system manipulations in anesthetized rats
Uploaded: 2018-09-11T14:00:00
Description: Watch this Scientific Journal Video about Quantifying Acute Changes in Renal Sympathetic Nerve Activity in Response to Central Nervous System Manipulations in Anesthetized Rats at JoVE.com

Questo studio è stato sostenuto dall'Istituto nazionale per la ricerca infermieristica (K99/R00NR014369).

Tutti i metodi descritti sono stati approvati dal Comitato istituzionale Animal Care presso l'Università dell'Illinois a Chicago.
1. creare elettrodi bipolari RSNA
<ol><li>Per creare l'elettrodo, tagliare due pezzi di filo di acciaio inossidabile lunghi circa 18 mm. Tagliare un pezzo di polietilene (PE-50) tubazione circa 15 mm di lunghezza. Alimentazione sia pezzi di filo nel tubo, lasciando il filo sporgente da entrambe le estremità.</li>
	<li>Togliere l'isolamento dalle estremità dei f...

<ol>
	<li>Mancia, G., Grassi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+autonomic+nervous+system+and+hypertension.">The autonomic nervous system and hypertension.</a> Circulation Research. 114 (11), 1804-1814 (2014).</li><li>Kannan, A., Medina, R. I., Nagajothi, N., Balamuthusamy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renal+sympathetic+nervous+system+and+the+effects+of+denervation+on+renal+arteries.">Renal sympathetic nervous system and the effects of denervation on renal arteries.</a> World Journal of Cardiology. 6 (8), 814-823 (2014).</li><li>Johns, E. J., Kopp, U. C., DiBona, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+control+of+renal+function.">Neural control of renal function.</a> Comprehensive Physiology. 1 (2), 767 (2011).</li><li>Fink, A. M., Dean, C., Piano, M. R., Carley, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pedunculopontine+tegmentum+controls+renal+sympathetic+nerve+activity+and+cardiorespiratory+activities+in+Nembutal-anesthetized+rats.">The pedunculopontine tegmentum controls renal sympathetic nerve activity and cardiorespiratory activities in Nembutal-anesthetized rats.</a> PLoS One. 12 (11), e0187956 (2017).</li><li>Dean, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endocannabinoid+modulation+of+sympathetic+and+cardiovascular+responses+to+acute+stress+in+the+periaqueductal+gray+of+the+rat.">Endocannabinoid modulation of sympathetic and cardiovascular responses to acute stress in the periaqueductal gray of the rat.</a> American Journal of Physiology, Regulatory, Integrative and Comparative Physiology. 300 (3), R771-R779 (2011).</li><li>Seagard, J. 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=Anandamide+content+and+interaction+of+endocannabinoid/GABA+modulatory+effects+in+the+NTS+on+baroreflex-evoked+sympathoinhibition.">Anandamide content and interaction of endocannabinoid/GABA modulatory effects in the NTS on baroreflex-evoked sympathoinhibition.</a> American Journal of Physiology-Heart and Circulatory Physiology. 286 (3), H992-H1000 (2004).</li><li>Ferreira, C. B., Cravo, S. L., Stocker, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Airway+obstruction+produces+widespread+sympathoexcitation:+Role+of+hypoxia,+carotid+chemoreceptors,+and+NTS+neurotransmission.">Airway obstruction produces widespread sympathoexcitation: Role of hypoxia, carotid chemoreceptors, and NTS neurotransmission.</a> Physiological Reports. 6 (3), (2018).</li><li>Stocker, S. D., Muntzel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+sympathetic+nerve+activity+chronically+in+rats:+Surgery+techniques,+assessment+of+nerve+activity,+and+quantification.">Recording sympathetic nerve activity chronically in rats: Surgery techniques, assessment of nerve activity, and quantification.</a> American Journal of Physiology-Heart and Circulatory Physiology. 305 (10), H1407-H1416 (2013).</li><li>Miki, K., Kosho, A., Hayashida, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+for+continuous+measurements+of+renal+sympathetic+nerve+activity+and+cardiovascular+function+during+exercise+in+rats.">Method for continuous measurements of renal sympathetic nerve activity and cardiovascular function during exercise in rats.</a> Experimental Physiology. 87 (1), 33-39 (2002).</li><li>Huetteman, D. A., Bogie, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+blood+pressure+monitoring+in+laboratory+rodents+via+implantable+radio+telemetry.">Direct blood pressure monitoring in laboratory rodents via implantable radio telemetry.</a> Methods in Molecular Biology. 573, 57-73 (2009).</li><li>Jespersen, B., Knupp, L., Northcott, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femoral+arterial+and+venous+catheterization+for+blood+sampling,+drug+administration+and+conscious+blood+pressure+and+heart+rate+measurements.">Femoral arterial and venous catheterization for blood sampling, drug administration and conscious blood pressure and heart rate measurements.</a> Journal of Visualized Experiments. (59), 3496 (2012).</li><li>Paxinos, G., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The rat brain in stereotaxic coordinates. , (2014).</li><li>Scislo, T. J., Augustyniak, R. A., O'Leary, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+arterial+baroreflex+regulation+of+renal,+lumbar,+and+adrenal+sympathetic+nerve+activity+in+the+rat.">Differential arterial baroreflex regulation of renal, lumbar, and adrenal sympathetic nerve activity in the rat.</a> American Journal of Physiology. 275, R995-R1002 (1998).</li><li>Kopp, U. C., Jones, S. Y., DiBona, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Afferent+renal+denervation+impairs+baroreflex+control+of+efferent+renal+sympathetic+nerve+activity.">Afferent renal denervation impairs baroreflex control of efferent renal sympathetic nerve activity.</a> American Journal of Physiology, Regulatory, Integrative and Comparative Physiology. 295 (6), R1882-R1890 (2008).</li><li>Guild, S. 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=Quantifying+sympathetic+nerve+activity:+problems,+pitfalls+and+the+need+for+standardization.">Quantifying sympathetic nerve activity: problems, pitfalls and the need for standardization.</a> Experimental Physiology. 95 (1), 41-50 (2010).</li><li>Stocker, S. D., Hunwick, K. J., Toney, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothalamic+paraventricular+nucleus+differentially+supports+lumbar+and+renal+sympathetic+outflow+in+water-deprived+rats.">Hypothalamic paraventricular nucleus differentially supports lumbar and renal sympathetic outflow in water-deprived rats.</a> Journal of Physiology. 15 (563 Pt 1), 249-263 (2005).</li><li>Stocker, S. D., Gordon, K. W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutamate+receptors+in+the+hypothalamic+paraventricular+nucleus+contribute+to+insulin-induced+sympathoexcitation.">Glutamate receptors in the hypothalamic paraventricular nucleus contribute to insulin-induced sympathoexcitation.</a> Neurophysiology. 113 (5), 1302-1309 (2015).</li><li>Saponjic, J., Radulovacki, M., Carley, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injection+of+glutamate+into+the+pedunculopontine+tegmental+nuclei+of+anesthetized+rat+causes+respiratory+dysrhythmia+and+alters+EEG+and+EMG+power.">Injection of glutamate into the pedunculopontine tegmental nuclei of anesthetized rat causes respiratory dysrhythmia and alters EEG and EMG power.</a> Sleep and Breathing. 9 (2), 82-91 (2005).</li><li>DiBona, G. F., Jones, S. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+analysis+of+renal+nerve+activity+responses+to+baroreceptor+denervation+in+hypertensive+rats.">Dynamic analysis of renal nerve activity responses to baroreceptor denervation in hypertensive rats.</a> Hypertension. 37 (4), 1153-1163 (2001).</li><li>Kunitake, T., Kannan, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discharge+pattern+of+renal+sympathetic+nerve+activity+in+the+conscious+rat:+spectral+analysis+of+integrated+activity.">Discharge pattern of renal sympathetic nerve activity in the conscious rat: spectral analysis of integrated activity.</a> Journal of Neurophysiology. 84 (6), 2859-2867 (2000).</li><li>Machado, B. H., Bonagamba, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microinjection+of+L-glutamate+into+the+nucleus+tractus+solitarii+increases+arterial+pressure+in+conscious+rats.">Microinjection of L-glutamate into the nucleus tractus solitarii increases arterial pressure in conscious rats.</a> Brain Research. 576 (1), 131-138 (1992).</li><li>Murakami, 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=Inhalation+anesthesia+is+preferable+for+recording+rat+cardiac+function+using+an+electrocardiogram.">Inhalation anesthesia is preferable for recording rat cardiac function using an electrocardiogram.</a> Biological and Pharmaceutical Bulletin. 37 (5), 834-839 (2014).</li><li>Nakamura, T., Tanida, M., Niijima, A., Hibino, H., Shen, J., Nagai, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auditory+stimulation+affects+renal+sympathetic+nerve+activity+and+blood+pressure+in+rats.">Auditory stimulation affects renal sympathetic nerve activity and blood pressure in rats.</a> Neuroscience Letters. 416 (2), 107-112 (2007).</li><li>Bardgett, M. E., McCarthy, J. J., Stocker, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutamatergic+receptor+activation+in+the+rostral+ventrolateral+medulla+mediates+the+sympathoexcitatory+response+to+hyperinsulinemia.">Glutamatergic receptor activation in the rostral ventrolateral medulla mediates the sympathoexcitatory response to hyperinsulinemia.</a> Hypertension. 55 (2), 284-290 (2010).</li><li>Brozoski, D. T., Dean, C., Hopp, F. A., Seagard, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+blockade+of+endocannabinoids+in+the+NTS+modulates+baroreflex-evoked+sympathoinhibition.">Uptake blockade of endocannabinoids in the NTS modulates baroreflex-evoked sympathoinhibition.</a> Brain Research. 1059 (2), 197-202 (2005).</li><li>Barman, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+can+we+learn+about+neural+control+of+the+cardiovascular+system+by+studying+rhythms+in+sympathetic+nerve+activity?.">What can we learn about neural control of the cardiovascular system by studying rhythms in sympathetic nerve activity?.</a> International Journal of Psychophysiology. 103, 69-78 (2016).</li><li>Charkoudian, N., Wallin, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sympathetic+neural+activity+to+the+cardiovascular+system:+integrator+of+systemic+physiology+and+interindividual+characteristics.">Sympathetic neural activity to the cardiovascular system: integrator of systemic physiology and interindividual characteristics.</a> Comprehensive Physiology. 4 (2), 825-850 (2014).</li><li>Malpas, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sympathetic+nervous+system+overactivity+and+its+role+in+the+development+of+cardiovascular+disease.">Sympathetic nervous system overactivity and its role in the development of cardiovascular disease.</a> Physiological Reviews. 90 (2), 513-557 (2010).</li><li>Chen, W. W., Xiong, X. Q., Chen, Q., Li, Y. H., Kang, Y. M., Zhu, G. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+sympathetic+afferent+reflex+and+its+implications+for+sympathetic+activation+in+chronic+heart+failure+and+hypertension.">Cardiac sympathetic afferent reflex and its implications for sympathetic activation in chronic heart failure and hypertension.</a> Acta Physiologica. 213 (4), 778-794 (2015).</li><li>Linz, 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=Modulation+of+renal+sympathetic+innervation:+Recent+insights+beyond+blood+pressure+control.">Modulation of renal sympathetic innervation: Recent insights beyond blood pressure control.</a> Clinical Autonomic Research. , (2018).</li></ol>

Fasi critiche per la misurazione RSNA includono: (1) evitando lo stretching dei nervi e dell'arteria renale durante la separazione del rene dal muscolo di paraspinal e quando viene posizionato il segmento del nervo gli elettrodi di registrazione, (2) attentamente sezionare le fibre nervose renale dal circostante tessuto/vaso, (3) assicurare che l'elettrodo fili sono privi di tessuto, sangue, o fluido linfatico e (4) impedendo il nervo di essiccazione mediante l'applicazione di olio minerale per il nervo renale ed il gel ...

Pre-cliniche scoperte circa controllo autonomo del sistema cardiovascolare informano le strategie per la gestione di patologie come ipertensione, insufficienza cardiaca e malattia renale cronica. Eccessiva attività del sistema nervoso simpatico e ridotto tono vagale cardiaco contribuire alla pressione sanguigna elevata (BP)1. Cronicamente elevato uscita simpatica renale migliora la secrezione di catecolamine e diminuisce il flusso ematico renale, con conseguenze deleterie per i sistemi cardiovascolari/renale2,3. Per definire le vie neurobiologiche che conducono alla disfunzione autonomica, studi nei roditori sono importanti per determinare come i neuroni del sistema nervoso centrale (SNC) regolano i parametri simpatici. Lo scopo del presente protocollo è quello di fornire informazioni tecniche su misura attività renale del nervo simpatico (RSNA) e BP e delineare le tecniche per quantificare i cambiamenti acuti simpatici in risposta a manipolazioni di CNS in ratti anestetizzati.
Misurazioni di acuta (non-sopravvivenza) RSNA (durata min a h) sono utili quando gli scienziati probe SNC farmacologicamente, elettricamente, o optogenetically in ratti per determinare le funzioni dei nuclei specifici anestetizzati. Utilizzando questi metodi, strutture come il nucleo solitario, gray periaqueductal, pedunculopontine tegmentum e rostral midollo ventrolateral sono stati studiati per definire vie neurobiologiche che regolano i parametri simpatico4, 5,6,7. Questo approccio è importante per identificare gli obiettivi di CNS per essere studiato ulteriormente nei modelli cronici di disfunzione autonomica8,9. Per completare questi esperimenti, il laboratorio richiede un saldatoio, microscopio operatorio, cornice stereotassica, amplificatore microelettrodo e monitor audio. A seconda dei fattori presenti nel laboratorio che contribuiscono al rumore elettrico, l'area chirurgica/registrazione può richiedere un Faraday gabbia/messa a terra cinturino per ridurre il rumore elettrico nella registrazione RSNA. Se analisi cervello richiederà il fissaggio del tessuto, un cappuccio di pompa e fumi di perfusione sono necessari. I dati possono essere digitalizzati e registrata utilizzando più dati fisiologici di software acquisizione (convertitore analogico-digitale) unità4,5, con opzioni di analisi diverse e compatibilità per l'incorporazione di segnali di telemetrici .

<table>
	<tbody>
		<tr>
			<td>Stainless steel wire</td>
			<td>A-M Systems; Sequim, WA</td>
			<td>791000</td>
			<td>RSNA electrode</td>
		</tr>
		<tr>
			<td>Polyethylene (PE-50) tubing</td>
			<td>VWR; Radnor, PA</td>
			<td>63019-048</td>
			<td>RSNA electrode; vessel cannulation</td>
		</tr>
		<tr>
			<td>Miniature pin connector</td>
			<td>A-M Systems; Sequim, WA</td>
			<td>520200</td>
			<td>RSNA electrode</td>
		</tr>
		<tr>
			<td>Crimping tool</td>
			<td>Daniels Manufacturing Corp.; Orlando, FL</td>
			<td>M22520</td>
			<td>RSNA electrode</td>
		</tr>
		<tr>
			<td>Connector strip</td>
			<td>Amphenol; Clinton Township, MI</td>
			<td>221-2653</td>
			<td>RSNA electrode</td>
		</tr>
		<tr>
			<td>J-B Kwik Epoxy</td>
			<td>J-B Weld, Sulphur Springs, TX</td>
			<td>8270</td>
			<td>RSNA electrode</td>
		</tr>
		<tr>
			<td>Silicone</td>
			<td>Permatex; Hartford, CT</td>
			<td>2222</td>
			<td>RSNA electrode</td>
		</tr>
		<tr>
			<td>Heparin sodium; Injectable (10 mL vial, 1000 U/mL)</td>
			<td>KV Veterinary Supply; David City, NE</td>
			<td>P03466</td>
			<td>Venous line patency</td>
		</tr>
		<tr>
			<td>Phenylephrine HCl; Injectable (1 mL vial; 10 mg/mL)</td>
			<td>ACE Surgical Supply; Brockton, MA</td>
			<td>950-6312</td>
			<td>Testing renal sympathoinhibition</td>
		</tr>
		<tr>
			<td>Single-hook elastic surgical stays</td>
			<td>Harvard Apparatus; Holliston, MA</td>
			<td>72-2595</td>
			<td>Incision</td>
		</tr>
		<tr>
			<td>Silk surgical tape</td>
			<td>3M, Minneapolis, MN</td>
			<td>1538-0</td>
			<td>Secure surgical stays</td>
		</tr>
		<tr>
			<td>Needles, 20 G</td>
			<td>Sigma-Aldrich; St. Louis, MO</td>
			<td>Z192554-100EA</td>
			<td>Vessel cannulation</td>
		</tr>
		<tr>
			<td>Dumont #7 curved forceps</td>
			<td>Fine Science Tools; Foster City, CA</td>
			<td>11274-20</td>
			<td>Vessel cannulation</td>
		</tr>
		<tr>
			<td>5-0 silk suture ties</td>
			<td>Braintree Scientific; Braintree, MA</td>
			<td>SUT-S 106</td>
			<td>Vessel cannulation</td>
		</tr>
		<tr>
			<td>Delicate hemostatic forceps</td>
			<td>Roboz Surgical Instrument Co.; Gaithersburg, MD</td>
			<td>RS-7117</td>
			<td>Vessel cannulation and RSNA surgery</td>
		</tr>
		<tr>
			<td>Crile Hemostatic forceps</td>
			<td>Fine Science Tools; Foster City, CA</td>
			<td>13004-14</td>
			<td>Needle bending</td>
		</tr>
		<tr>
			<td>Telemetry transmitter</td>
			<td>Data Sciences International; Minneapolis, MN</td>
			<td>PA-10</td>
			<td>Mean arterial pressure monitoring (telemetry)</td>
		</tr>
		<tr>
			<td>Re-gel syringe</td>
			<td>Data Sciences International; Minneapolis, MN</td>
			<td>276-0038-001</td>
			<td>Transmitter reuse (telemetry)</td>
		</tr>
		<tr>
			<td>Disposable pressure transducer</td>
			<td>Transpac; San Clemente, CA</td>
			<td>MI-1224</td>
			<td>Mean arterial pressure monitoring</td>
		</tr>
		<tr>
			<td>Clear-Cuff pressure infuser</td>
			<td>MILA International Inc.; Florence, KY</td>
			<td>2281339</td>
			<td>Mean arterial pressure monitoring</td>
		</tr>
		<tr>
			<td>Vessel cannulation forceps</td>
			<td>Fine Science Tools; Foster City, CA</td>
			<td>00574-11</td>
			<td>Catheter insertion</td>
		</tr>
		<tr>
			<td>Black monofilament nylon 4-0 suture on reverse cutting needle</td>
			<td>McKesson Medical-Surgical; San Francisco, CA</td>
			<td>S661GX</td>
			<td>Secure telemetry transmitter</td>
		</tr>
		<tr>
			<td>Telemetry receiver</td>
			<td>Data Sciences International; Minneapolis, MN</td>
			<td>RPC-1</td>
			<td>Mean arterial pressure monitoring (telemetry)</td>
		</tr>
		<tr>
			<td>LabChart Pro (software), PowerLab (acquisition hardware)</td>
			<td>AD Instruments; Colorado Springs, CO</td>
			<td>ML846, MX2 matrix 2.0 (Compatible with Data Science International telemetry)</td>
			<td>3 options for software/acquisition hardware</td>
		</tr>
		<tr>
			<td>SciWorks (software), DataWave (acquisition hardware)</td>
			<td>DataWave Technologies, Loveland, CO</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Spike 2 (software), Micro1401-3</td>
			<td>Cambridge Electronic Design Ltd., London UK</td>
			<td>1401-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-drill</td>
			<td>Roboz Surgical Instrument Co.; Gaithersburg, MD</td>
			<td>RS-6300</td>
			<td>CNS surgery</td>
		</tr>
		<tr>
			<td>Stereotaxic surgery frame</td>
			<td>Stoelting; Wood Dale, IL</td>
			<td>51600</td>
			<td>CNS surgery</td>
		</tr>
		<tr>
			<td>Microelectrode amplifier with 10X pre-amplifier</td>
			<td>A-M Systems; Sequim, WA</td>
			<td>1800-2</td>
			<td>RSNA recording</td>
		</tr>
		<tr>
			<td>Retractors</td>
			<td>Fine Science Tools; Foster City, CA</td>
			<td>17009-07</td>
			<td>RSNA surgery</td>
		</tr>
		<tr>
			<td>Micro-dissecting tweezers</td>
			<td>Fine Science Tools; Foster City, CA</td>
			<td>11251-10</td>
			<td>RSNA surgery</td>
		</tr>
		<tr>
			<td>Micro-hook</td>
			<td>Fine Science Tools; Foster City, CA</td>
			<td>10064-14</td>
			<td>RSNA surgery</td>
		</tr>
		<tr>
			<td>Mineral oil</td>
			<td>Fisher Scientific; Waltham, MA</td>
			<td>8042-47-5</td>
			<td>RSNA surgery</td>
		</tr>
		<tr>
			<td>Audio monitor</td>
			<td>A-M Systems; Sequim, WA</td>
			<td>3300</td>
			<td>RSNA surgery</td>
		</tr>
		<tr>
			<td>Silica gel</td>
			<td>Wacker, Munchen; Germany</td>
			<td>RT601A-B</td>
			<td>RSNA surgery</td>
		</tr>
		<tr>
			<td>Electrical clips</td>
			<td>Tyco Electronics; Schaffhausen, Switzerland</td>
			<td>EB0283-000</td>
			<td>Grounding or securing perfusion needle</td>
		</tr>
		<tr>
			<td>Bonn scissors, straight/sharp points</td>
			<td>Roboz Surgical Instrument Co; Gaithersburg, MD</td>
			<td>RS-5840</td>
			<td>Perfusion</td>
		</tr>
		<tr>
			<td>Gavage needle</td>
			<td>Harvard Apparatus; Holliston, MA</td>
			<td>75-0286</td>
			<td>Perfusion</td>
		</tr>
		<tr>
			<td>Masterflex perfusion pump</td>
			<td>Cole-Parmer; Vernon Hills, IL</td>
			<td>7524-10</td>
			<td>Perfusion</td>
		</tr>
		<tr>
			<td>Masterflex platinum-cured silicone tubing</td>
			<td>Cole-Parmer; Vernon Hills, IL</td>
			<td>96410-15</td>
			<td>Perfusion</td>
		</tr>
		<tr>
			<td>Formalin (10% buffered solution; 4 L)</td>
			<td>Sigma-Aldrich; St. Louis, MO</td>
			<td>HT501128</td>
			<td>Perfusion</td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma-Aldrich; St. Louis, MO</td>
			<td>S0389</td>
			<td>Cryoprotection</td>
		</tr>
	</tbody>
</table>

quantifying acute changes in renal sympathetic nerve activity in response to central nervous system manipulations in anesthetized rats

Attività renale del nervo simpatico (RSNA) e la pressione arteriosa media sono parametri importanti nella ricerca cardiovascolare ed autonoma; Tuttavia, ci sono risorse limitate dirigere gli scienziati nelle tecniche per la misurazione e l'analisi di queste variabili. Questo protocollo descrive i metodi per la misurazione RSNA e pressione arteriosa media in ratti anestetizzati. Il protocollo comprende anche gli approcci per l'accesso al cervello durante le registrazioni di RSNA per manipolazioni del sistema nervoso centrale (SNC). La tecnica di registrazione RSNA è compatibile con farmacologico, optogenetica, o stimolazione elettrica dello SNC. L'approccio è utile quando un investigatore si misura a breve termine (min a h) le risposte autonomiche in esperimenti non-sopravvivenza per correlare anatomicamente con nuclei di CNS. L'approccio non è destinato a essere utilizzato per ottenere registrazioni cronica (sopravvivenza) di RSNA in ratti. Gli scarichi in RSNA, una media rettificata RSNA, e la pressione arteriosa media può essere quantificato e analizzati ulteriormente usando test statistici parametrici. Metodi per ottenere accesso venoso, la pressione arteriosa media di registrazione telemetrically e la fissazione di cervello per future analisi istologica sono anche descritti nell'articolo.

La figura 1 illustra un'esempio RSNA e BP la registrazione da un ratto anestetizzato Nembutal. Un'iniezione endovenosa di fenilefrina è stata utilizzata per indurre un aumento nella pressione arteriosa media e per evocare il baroriflesso e transitorio sympathoinhibition4,6. Per quantificare RSNA, il crudo RSNA è stato rettificato e in media per non sovrapposte 10 segmenti di s; la stima del rumore ?...

Watch this Scientific Journal Video about Quantifying Acute Changes in Renal Sympathetic Nerve Activity in Response to Central Nervous System Manipulations in Anesthetized Rats at JoVE.com

Quantifying Acute Changes in Renal Sympathetic Nerve Activity in Response to Central Nervous System Manipulations in Anesthetized Rats

Quantificare i cambiamenti acuti nell'attività comprensiva renale del nervo in risposta a manipolazioni del sistema nervoso centrale in ratti anestetizzati

Metodi per misurare le risposte cardiovascolari e simpatiche a manipolazioni del sistema nervoso centrale (SNC) sono importanti per far progredire la neuroscienza. Questo protocollo è stato sviluppato per aiutare gli scienziati con misurare e quantificare i cambiamenti acuti nell'attività comprensiva renale del nervo (RSNA) in ratti anestetizzati (non-sopravvivenza).

Renal sympathetic nerve activity (RSNA) and mean arterial pressure are important parameters in cardiovascular and autonomic research; however, there are ...

Research

JoVE Journal

Neuroscience

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

Resezione del nervo ottico: un modello di apoptosi Neuron adulti nel Sistema Nervoso Centrale

Retinal ganglion cells (RGCs) are CNS neurons that output visual information from the retina to the brain, via the optic nerve. 
The optic nerve can be ...

Ricerca

Neuroscienze

Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice

Imaging multimodale impianto di cellule staminali nel sistema nervoso centrale di topi

During the past decade, stem cell transplantation has gained increasing interest as primary or secondary therapeutic modality for a variety of diseases, ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Due registrazioni elettrofisiologiche di sinapticamente-evocati Risposte astrogliale e neuronale in acute fettine di ippocampo

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

Multi-unit Metodi di registrazione per caratterizzare l&#39;attività neurale nel Locust (<em&gt; Schistocerca Americana</em&gt;) Circuiti olfattive

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

Etichettatura di singole cellule nel sistema nervoso centrale di<em&gt; Drosophila melanogaster</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

An Injury Paradigm to Investigate Central Nervous System Repair in Drosophila

An Injury Paradigm to Investigate Central Nervous System Repair in Drosophila

Un paradigma Infortunio per Indagare riparazione del sistema nervoso centrale in<em&gt; Drosophila</em

An experimental method has been developed to investigate the cellular responses to central nervous system (CNS) injury using the fruit-fly Drosophila. ...

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

Consegna diretta intraventricolare di farmaci per il sistema nervoso centrale, Roditore

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

Immunohistochemical Analysis in the Rat Central Nervous System and Peripheral Lymph Node Tissue Sections

Analisi immunoistochimica nel Sistema Nervoso Centrale e Periferico Rat linfa nodo del tessuto Sezioni

Immunohistochemistry (IHC) provides highly specific, reliable and attractive protein visualization. Correct performance and interpretation of an IHC-based ...

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

In vivo Interrogatorio del Sistema Nervoso Centrale Translatome da Polyribosome Frazionamento

Multiple processes are involved in gene expression including transcription, translation and stability of mRNAs and proteins. Each of these steps are ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo optogenetic stimolazione del sistema nervoso centrale Roditore

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...