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

Denna studie stöddes av National Institute for Nursing Research (K99/R00NR014369).

Alla metoder beskrivs godkändes av den institutionella djur eftervård kommittén vid University of Illinois at Chicago.
1. skapa bipolär RSNA elektroder
<ol><li>För att skapa elektroden, skär två bitar av tråd av rostfritt stål varje ca 18 mm långa. Skär en bit av polyeten (PE-50) slang ca 15 mm lång. Foder båda bitar av tråd in slangen, lämnar tråden sticker ut från båda ändar.</li>
	<li>Ta bort isoleringen från ändarna på kablarna; trimma ledningarna lämnar 2-3 mm av ...

<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>

Kritiska steg för att mäta RSNA inkluderar: (1) att undvika stretching av Njurartären och nerver när separera njuren från paraspinal muskeln och när du placerar segmentet nerv på inspelning elektroderna, (2) noggrant dissekera nedsatt nervfibrerna från omgivande vävnad/fartyget, (3) säkerställa att elektroden ledningar är fria från vävnad, blod, eller lymfan vätska och (4) hindrar nerven från att torka ut genom att använda mineralolja nedsatt nerv och kiselgel till nerv-elektrod enheten. För felsökning...

Prekliniska upptäckter om autonom kontroll av det kardiovaskulära systemet informera strategier för att hantera störningar såsom hypertoni, hjärtsvikt och kronisk njursjukdom. Överaktivitet av sympatiska nervsystemet och minskad vagala hjärt tonen bidra till förhöjt blodtryck (BP)1. Kroniskt förhöjda nedsatt sympatiska utflöde förbättrar katekolamin sekretion och minskar renalt blodflöde, med skadliga konsekvenser för hjärt-/ njurarna2,3. För att definiera den neurobiologiska vägar som leder till autonom dysfunktion, är studier på gnagare viktiga för att fastställa hur centrala nervsystemet (CNS) nervceller reglera sympatiska parametrar. Syftet med detta protokoll är att tillhandahålla teknisk information om att mäta njurfunktion sympatisk nervaktivitet (RSNA) och BP och att beskriva teknikerna för att kvantifiera akut sympatiska förändringar i svar till CNS manipulationer hos sövda råttor.
Akut (icke-överlevnad) RSNA mätningar (varaktig min h) är användbara när forskare kommer probe CNS farmakologiskt, elektriskt, eller optogenetically i sövda råttor att bestämma specifika kärnor funktioner. Med dessa metoder, strukturer såsom ensam kärnan, periaqueductal gray, har pedunculopontine tegmentum, och rostralt ventrolaterala medulla undersökts för att definiera neurobiologiska vägar reglera sympatiska parametrar4, 5,6,7. Denna strategi är viktig för att identifiera CNS mål att undersökas ytterligare i kronisk modeller av autonom dysfunktion8,9. För att slutföra dessa experiment, kräver laboratoriet en lödkolv, kirurgiska Mikroskop, stereotaxic ram, mikroelektrod förstärkare och ljud monitor. Beroende på vilka faktorer i laboratoriet som bidrar till elektriska störningar, kan området kirurgiska/inspelning kräva en Faradays bur/jordning rem att minska elektriskt brus i RSNA inspelningen. Om hjärnan analyser kommer att kräva vävnad fixering, krävs en perfusion pump och rök huva. Data kan digitaliseras och registreras med flera fysiologiska data programvara/förvärv (analog-digital converter) enheter4,5, med olika analysalternativ och kompatibla för att införliva telemetriska signaler .

<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

Nedsatt sympatisk nervaktivitet (RSNA) och genomsnittliga arteriella trycket är viktiga parametrar i hjärt- och autonoma forskning; Det finns dock begränsade resurser rikta forskare i tekniker för att mäta och analysera dessa variabler. Det här protokollet beskriver metoder för att mäta RSNA och genomsnittliga arteriella trycket hos sövda råttor. Protokollet innehåller också metoderna för åtkomst till hjärnan under RSNA inspelningar för centrala nervsystemet (CNS) manipulationer. RSNA inspelning tekniken är kompatibel med farmakologiska, optogenetic, eller elektrisk stimulering av centrala nervsystemet. Metoden är användbar när en utredare kommer att mäta kortsiktiga (min till h) autonoma svaren i icke-survival experiment att korrelera anatomiskt med CNS atomkärnor. Metoden är inte avsett att användas för att få kronisk (överlevnad) inspelningar av RSNA hos råttor. Utsläpp i RSNA, i genomsnitt rättas RSNA och genomsnittliga arteriella trycket kan kvantifieras och analyseras ytterligare med parametriska statistiska tester. Metoder för att få venös access, inspelning genomsnittliga arteriella trycket telemetrically och hjärnan fixering för framtida histologisk analys beskrivs också i artikeln.

Figur 1 illustrerar en prov RSNA och BP inspelning från en Nembutal-sövd råtta. En intravenös injektion av fenylefrin användes att inducera en ökning av medelvärdet artärtryck och att framkalla baroreflex och övergående sympathoinhibition4,6. För att kvantifiera RSNA, var den råa RSNA rättas och ett medelvärde för icke-överlappande 10 s segment; buller uppskattningen var subtraheras fr...

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

Kvantifiera akuta förändringar i Renal sympatisk nervaktivitet svar på centrala nervsystemet manipulationer hos sövda råttor

Metoder för mätning av sympatiska och kardiovaskulära Svaren till centrala nervsystemet (CNS) manipulationer är viktiga för att främja neurovetenskap. Detta protokoll har utvecklats för att hjälpa forskare med att mäta och kvantifiera akuta förändringar i renal sympatisk nervaktivitet (RSNA) hos sövda råttor (icke-överlevnad).

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

Research

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

Synnerven transection: En modell av Vuxen Neuron apoptos i det centrala nervsystemet

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 ...

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

Multimodal Imaging av stamceller implanteras i centrala nervsystemet hos möss

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

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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-enhet inspelning Metoder för att karaktärisera neural aktivitet i Locust (<em&gt; Schistocerca Americana</em&gt;) Olfactory kretsar

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

Märkning av enstaka celler i det centrala nervsystemet hos<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

En skada Paradigm att undersöka centrala nervsystemet Repair i<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

Direkt Intraventrikulär tillförsel av läkemedel till Gnagare Centrala nervsystemet

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

Immunohistokemisk analys i Rat centrala nervsystemet och perifera lymfkörtelvävnadssnitt

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 Förhör av centrala nervsystemet Translatome av Polyribosome Fractionation

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 Stimulering av gnagare centrala nervsystemet

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