Name: paired whole cell recordings in organotypic hippocampal slices
Uploaded: 2014-09-28T14:00:00
Description: Watch this Scientific Journal Video about Paired Whole Cell Recordings in Organotypic Hippocampal Slices at JoVE.com

We would like to thank the members of the Montgomery and Madison labs for helpful discussion. We acknowledge the funding received from the following sources in this research: NFNZ, AMRF, Marsden Fund, HRC, and NIH.

पशु आचार विवरण: इस पांडुलिपि में वर्णित प्रोटोकॉल ऑकलैंड और स्टैनफोर्ड विश्वविद्यालय के विश्वविद्यालय द्वारा स्थापित जानवरों की देखभाल के दिशा निर्देशों का पालन करें. P7 चूहे पिल्ले तेजी ?...

<ol>
	<li>Dingledine, R., Borges, K., Bowie, D., Traynelis, S. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+glutamate+receptor+ion+channels.">The glutamate receptor ion channels.</a> Pharmacology Reviews. 51, 7-61 (1999).</li><li>Malenka, R. C., Nicoll, R. A. <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+potentiation+-+A+decade+of+progress.">Long-term potentiation - A decade of progress.</a> Science. 285, 1870-1874 (1999).</li><li>Bliss, T. V. P., Lomo, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-lasting+potentiation+of+synaptic+transmission+in+the+dentate+area+of+the+anaesthetized+rabbit+following+stimulation+of+the+perforant+path.">Long-lasting potentiation of synaptic transmission in the dentate area of the anaesthetized rabbit following stimulation of the perforant path.</a> Journal of Physiology. 232, 331-356 (1973).</li><li>Dudek, S. M., Bear, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homosynaptic+long-term+depression+in+area+CA1+of+hippocampus+and+effects+of+N-methyl-D-aspartate+receptor+blockade.">Homosynaptic long-term depression in area CA1 of hippocampus and effects of N-methyl-D-aspartate receptor blockade.</a> Proceedings of the National Academy of Sciences USA. 89, 4363-4367 (1992).</li><li>Borst, J. G., Soria van Hoeve, 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+calyx+of+held+synapse:+from+model+synapse+to+auditory+relay.">The calyx of held synapse: from model synapse to auditory relay.</a> Annual Reviews in Physiology. 74, 199-224 (2012).</li><li>Pavlidis, P., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+transmission+in+pair+recordings+from+CA3+pyramidal+cells+in+organotypic+culture.">Synaptic transmission in pair recordings from CA3 pyramidal cells in organotypic culture.</a> Journal of Neurophysiology. 81, 2787-2797 (1999).</li><li>Pavlidis, P., Montgomery, J. M., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presynaptic+protein+kinase+activity+supports+long-term+potentiation+at+synapses+between+individual+hippocampal+neurons.">Presynaptic protein kinase activity supports long-term potentiation at synapses between individual hippocampal neurons.</a> Journal of Neuroscience. 20 (12), 4497-4505 (2000).</li><li>Montgomery, J. M., Pavlidis, P., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pair+recordings+reveal+all-silent+synaptic+connections+and+the+postsynaptic+expression+of+long-term+potentiation.">Pair recordings reveal all-silent synaptic connections and the postsynaptic expression of long-term potentiation.</a> Neuron. 29, 691-701 (2001).</li><li>Montgomery, J. M., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=State-dependent+heterogeneity+in+synaptic+depression+between+pyramidal+cell+pairs.">State-dependent heterogeneity in synaptic depression between pyramidal cell pairs.</a> Neuron. 33, 765-777 (2002).</li><li>Montgomery, J. M., Selcher, J. C., Hansen, J. E., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamin-dependent+NMDAR+endocytosis+during+LTD+and+its+dependence+on+synaptic+state.">Dynamin-dependent NMDAR endocytosis during LTD and its dependence on synaptic state.</a> BMC Neuroscience. 6, 48 (2005).</li><li>Waites, C. 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=Synaptic+SAP97+isoforms+regulate+AMPA+receptor+dynamics+and+access+to+presynaptic+glutamate.">Synaptic SAP97 isoforms regulate AMPA receptor dynamics and access to presynaptic glutamate.</a> Journal of Neuroscience. 29 (14), 4332-4345 (2009).</li><li>Emond, 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=AMPA+receptor+subunits+define+properties+of+state-dependent+synaptic+plasticity.">AMPA receptor subunits define properties of state-dependent synaptic plasticity.</a> Journal of Physiology. 588, 1929-1946 (2010).</li><li>Li, 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=SAP97+directs+NMDA+receptor+spine+targeting+and+synaptic+plasticity.">SAP97 directs NMDA receptor spine targeting and synaptic plasticity.</a> Journal of Physiology. 589, 4491-4510 (2011).</li><li>Genoux, D., Bezerra, P., Montgomery, 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=Intra-spaced+stimulation+and+protein+phosphatase+1+dictate+the+direction+of+synaptic+plasticity.">Intra-spaced stimulation and protein phosphatase 1 dictate the direction of synaptic plasticity.</a> European Journal of Neuroscience. 33 (10), 1761-1770 (2011).</li><li>Selcher, J. C., Xu, W., Hanson, J. E., Malenka, R. C., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glutamate+receptor+subunit+GluA1+is+necessary+for+long-term+potentiation+and+synapse+unsilencing,+but+not+long-term+depression+in+mouse+hippocampus.">Glutamate receptor subunit GluA1 is necessary for long-term potentiation and synapse unsilencing, but not long-term depression in mouse hippocampus.</a> Brain Research. 1435, 8-14 (2012).</li><li>Arons, M. 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=Autism-associated+mutations+in+ProSAP2/Shank3+impair+synaptic+transmission+and+neurexin-neuroligin-mediated+transsynaptic+signaling.">Autism-associated mutations in ProSAP2/Shank3 impair synaptic transmission and neurexin-neuroligin-mediated transsynaptic signaling.</a> Journal of Neuroscience. 32 (43), 14966-14978 (2012).</li><li>Montgomery, J. M., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discrete+synaptic+states+define+a+major+mechanism+of+synapse+plasticity.">Discrete synaptic states define a major mechanism of synapse plasticity.</a> Trends in Neuroscience. 27 (12), 744-750 (2004).</li><li>Miles, R., Poncer, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pair+recordings+from+neurones.">Pair recordings from neurones.</a> Current Opinion in Neurobiology. 6 (3), 387-394 (1996).</li><li>Malinow, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+between+pairs+of+hippocampal+slice+neurons:+quantal+levels,+oscillations+and+LTP.">Transmission between pairs of hippocampal slice neurons: quantal levels, oscillations and LTP.</a> Science. 252 (5006), 722-724 (1991).</li><li>G&#228;hwiler, B. H., Capogna, M., Debanne, D., McKinney, R. A., Thompson, 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=Organotypic+slice+cultures:+a+technique+has+come+of+age.">Organotypic slice cultures: a technique has come of age.</a> Trends in Neuroscience. 20 (10), 471-477 (1997).</li><li>Stoppini, L., Buchs, P. A., Muller, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+for+organotypic+cultures+of+nervous+tissue.">A simple method for organotypic cultures of nervous tissue.</a> Journal of Neuroscience Methods. 37 (2), 173-182 (1991).</li><li>Debanne, D., Gu&#233;rineau, N. C., G&#228;hwiler, B. H., Thompson, 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=Paired-pulse+facilitation+and+depression+at+unitary+synapses+in+rat+hippocampus:+quantal+fluctuation+affects+subsequent+release.">Paired-pulse facilitation and depression at unitary synapses in rat hippocampus: quantal fluctuation affects subsequent release.</a> Journal of Physiology. 491, 163-176 (1996).</li><li>Debanne, D., G&#228;hwiler, B. H., Thompson, 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=Cooperative+interactions+in+the+induction+of+long-term+potentiation+and+depression+of+synaptic+excitation+between+hippocampal+CA3-CA1+cell+pairs+in+vitro.">Cooperative interactions in the induction of long-term potentiation and depression of synaptic excitation between hippocampal CA3-CA1 cell pairs in vitro.</a> Proceedings of the National Academy of Sciences U S A. 93 (20), 11225-11230 (1996).</li><li>Malinow, R., Tsien, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presynaptic+enhancement+shown+by+whole+cell+recordings+of+long-term+potentiation+in+hippocampal+slices.">Presynaptic enhancement shown by whole cell recordings of long-term potentiation in hippocampal slices.</a> Nature. 346 (6280), 177-180 (1990).</li><li>DeBello, W. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SNAP-mediated+protein-protein+interactions+essential+for+neurotransmitter+release.">SNAP-mediated protein-protein interactions essential for neurotransmitter release.</a> Nature. 373 (6515), 626-630 (1995).</li><li>Bekkers, J. M., Stevens, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Origin+of+variability+in+quantal+size+in+cultured+hippocampal+neurons+and+hippocampal+slices.">Origin of variability in quantal size in cultured hippocampal neurons and hippocampal slices.</a> Proceedings of the National Academy of Sciences U S A. 87, 5359-5362 (1990).</li><li>Malinow, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+between+pairs+of+hippocampal+slice+neurons:+quantal+levels,+oscillations,+and+LTP.">Transmission between pairs of hippocampal slice neurons: quantal levels, oscillations, and LTP.</a> Science. 252, 722-724 (1991).</li><li>Debanne, D., Gu&#233;rineau, N. C., G&#228;hwiler, B. H., Thompson, 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=Physiology+and+pharmacology+of+unitary+synaptic+connections+between+pairs+of+cells+in+areas+CA3+and+CA1+of+rat+hippocampal+slice+cultures.">Physiology and pharmacology of unitary synaptic connections between pairs of cells in areas CA3 and CA1 of rat hippocampal slice cultures.</a> Journal of Neurophysiology. 73 (3), 1282-1294 (1995).</li><li>Mitra, A., Blank, M., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmentally+altered+inhibition+in+Ts65Dn,+a+mouse+model+of+Down+syndrome.">Developmentally altered inhibition in Ts65Dn, a mouse model of Down syndrome.</a> Brain Research. 1440, 1-8 (2012).</li><li>Hanson, J. E., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presynaptic+FMR1+genotype+influences+the+degree+of+synaptic+connectivity+in+a+mosaic+mouse+model+of+fragile+X+syndrome.">Presynaptic FMR1 genotype influences the degree of synaptic connectivity in a mosaic mouse model of fragile X syndrome.</a> Journal of Neuroscience. 27 (15), 4014-4018 (2007).</li><li>Hanson, J. E., Blank, M., Valenzuela, R. A., Garner, C. C., Madison, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+functional+nature+of+synaptic+circuitry+is+altered+in+area+CA3+of+the+hippocampus+in+a+mouse+model+of+Down's+syndrome.">The functional nature of synaptic circuitry is altered in area CA3 of the hippocampus in a mouse model of Down's syndrome.</a> Journal of Physiology. 579, 53-67 (2007).</li></ol>

यहाँ हम organotypic hippocampal टुकड़ा संस्कृतियों में सफल बनती पूरे सेल रिकॉर्डिंग की स्थापना के लिए आवश्यकताओं का वर्णन किया है. बनती रिकॉर्डिंग भी तीव्र स्लाइस और अलग संस्कृति प्रणालियों 26,27 सहित कई तैयारियो...

ग्लूटामेट रिसेप्टर्स केंद्रीय तंत्रिका तंत्र synapses पर उत्तेजक synaptic प्रसारण के बहुमत मध्यस्थता. पोस्टअन्तर्ग्रथनी झिल्ली की रीढ़ सिर पर स्थानीयकृत ionotropic ग्लूटामेट रिसेप्टर्स की दो प्रमुख उपप्रकार एन मिथाइल-D-aspartate (NMDA) और α-अमीनो-3-hydroxy-5 proprionic-4-methylisoxazole एसिड (AMPA) रिसेप्टर्स हैं. झिल्ली क्षमता आराम से कम, AMPA रिसेप्टर्स synaptic प्रसारण के दौरान पोस्टअन्तर्ग्रथनी वर्तमान के सबसे ले. हिप्पोकैम्पस में, NMDA रिसेप्टर पोस्टअन्तर्ग्रथनी झिल्ली में AMPA रिसेप्टर्स की संख्या में परिवर्तन ट्रिगर करने में एक महत्वपूर्ण भूमिका निभाता है: एक &quot;संयोग डिटेक्टर&quot; के रूप में अभिनय द्वारा 1 अन्तर्ग्रथनी शक्ति 1 में परिवर्तन शुरू करने के लिए, NMDA रिसेप्टर synaptic तंत्र में भाग लेता है कि एक subcellular स्तर पर सीखने और स्मृति पिन लगा रहे हैं. Presynaptic ट्रांसमीटर रिलीज के साथ समानांतर में पोस्टअन्तर्ग्रथनी न्यूरॉन की विध्रुवण के जवाब में, कैल्शियम NMDA के माध्यम से प्रवेश करती हैरिसेप्टर AMPA रिसेप्टर प्रविष्टि या हटाने 2 आरंभ करने के लिए. ये रिसेप्टर गतिशीलता अन्तर्ग्रथन plasticity आबाद: अन्तर्ग्रथनी शक्ति में कमी लंबे समय तक अवसाद 4 (लिमिटेड) है, जबकि अन्तर्ग्रथनी शक्ति में वृद्धि हुई है, लंबी अवधि potentiation 2,3 (एलटीपी) है. NMDA रिसेप्टर्स अपने प्रेरण नियंत्रित करने के लिए लगा रहे हैं, जबकि इसलिए AMPA रिसेप्टर आंदोलन, synaptic plasticity अभिव्यक्ति के लिए जिम्मेदार माना जाता है. Synaptic प्रसारण और plasticity अंतर्निहित सटीक तंत्र का निर्धारण आदर्श synapses के छोटे आबादी, एकल synapses का अध्ययन की आवश्यकता है. कुछ synapses अत्यधिक होने के कारण synaptic कनेक्शन के छोटे और फैलाना प्रकृति के इस बेहद मुश्किल है सबसे synaptic आबादी के लिए इस स्तर, उदाहरण के लिए, 5 आयोजित की बाह्यदलपुंज, पर अध्ययन के लिए अनुकूल हैं जबकि. दो प्रमुख इलेक्ट्रोफिजियोलॉजी तकनीक एकल synaptic कनेक्शन की जांच करने के लिए विकसित किया गया है: पहले न्यूनतम उत्तेजना है, wherएक प्रीसानेप्टिक फाइबर माना जाता है ई extracellularly को प्रेरित किया. दूसरी तकनीक synaptically जुड़े न्यूरॉन्स से दो एक साथ पूरे सेल रिकॉर्डिंग किया जाता है जहां रिकॉर्डिंग, बनती है. कम से कम उत्तेजना का एक बड़ा फायदा यह एक साथ एक पोस्टअन्तर्ग्रथनी न्यूरॉन से रिकॉर्डिंग जबकि axonal पथ में एक बाह्य उत्तेजक इलेक्ट्रोड की नियुक्ति से जुड़े, प्रदर्शन करने के लिए तेजी से और अपेक्षाकृत सरल है. इस तकनीक का उपयोग करते समय प्राथमिक चिंता का विषय एक एकल कक्ष के विश्वसनीय उत्तेजना शायद ही कभी परीक्षण के बाद परीक्षण गारंटी दी जा सकती है. पिछले पंद्रह वर्षों से हम नियमित तौर पर दो synaptically जुड़े पिरामिड न्यूरॉन्स 6-17 से पूरे सेल रिकॉर्डिंग बनती इस्तेमाल किया है. इस तकनीक का प्रमुख लाभ केवल एक प्रीसानेप्टिक न्यूरॉन लगातार और मज़बूती से प्रेरित है. यह भी electrophysiological लक्षण वर्णन भी है लेकिन प्रीसानेप्टिक न्यूरॉन 6,18 &lt;के औषधीय हेरफेर न केवल अनुमति देता है/ Sup&gt;. हालांकि, न्यूरॉन्स के बीच synaptic कनेक्टिविटी की संभावना 19 प्राप्त करने के लिए जुड़ा जोड़े मुश्किल बना, कम है. organotypic मस्तिष्क टुकड़ा संस्कृतियों का उपयोग synaptic संपर्क के रूप में इस बाधा विट्रो में फिर से स्थापित कर सकते हैं और इसके अलावा परिणामस्वरूप कनेक्टिविटी की प्रकृति देशी मस्तिष्क ऊतक 20 में समान है circumvents. इसके अलावा, organotypic संस्कृतियों को प्लास्टिसिटी तंत्र को सक्षम करने, एलटीपी, लिमिटेड 7-10,12-15,21 और बनती पल्स सुविधा (पीपीएफ) और अवसाद (पीपीडी) 6,22,23 सहित अल्पकालिक synaptic plasticity के अतिरिक्त रूपों व्यक्त न्यूरॉन्स के जोड़े में अध्ययन किया. यहाँ हम इस सफलतापूर्वक इन विट्रो प्रणाली में बनती रिकॉर्डिंग को प्राप्त करने में शामिल विस्तृत कार्यप्रणाली का वर्णन. यह जानकारी आसानी से तीव्र स्लाइस और अन्य मस्तिष्क क्षेत्रों सहित अन्य प्रयोगात्मक सिस्टम, के लिए अनुकूलित किया जा सकता है.

<table border="0" cellpadding="0" cellspacing="0" width="686">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:256px;">Organotypic cultures</td>
			<td style="width:111px;"></td>
			<td style="width:255px;"></td>
			<td style="width:64px;">Paired recordings</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Minimum Essential Medium</td>
			<td></td>
			<td></td>
			<td>Stable motorized micromanipulators&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Penicillin-Streptomycin solution&#160;</td>
			<td></td>
			<td></td>
			<td>Shallow tissue bath</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HEPES buffer solution</td>
			<td></td>
			<td></td>
			<td>DIC camera</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1M Tris stock solution&#160;</td>
			<td></td>
			<td></td>
			<td>Amplifier</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hank&#8217;s Balanced Salt Solution</td>
			<td></td>
			<td></td>
			<td>Computer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Horse Serum&#160;</td>
			<td></td>
			<td></td>
			<td>Vibration isolation table</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">plastic-coated miniature spatulas&#160;</td>
			<td></td>
			<td></td>
			<td>Upright microscope</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">soft paintbrush&#160;</td>
			<td></td>
			<td></td>
			<td>Data acquistion and analysis software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">manual tissue chopper</td>
			<td></td>
			<td></td>
			<td>Electrode puller</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">#2 filter paper</td>
			<td></td>
			<td></td>
			<td>Faraday cage</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">#5&#160; forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Membrane inserts</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">CO2 incubator&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissection hood</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Class II hood</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

paired whole cell recordings in organotypic hippocampal slices

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct electrophysiological characterization of the synaptic connections between individual neurons, but also pharmacological manipulation of either the presynaptic or the postsynaptic neuron. When carried out in organotypic hippocampal slice cultures, the probability that two neurons are synaptically connected is significantly increased. This preparation readily enables identification of cell types, and the neurons maintain their morphology and properties of synaptic function similar to that in native brain tissue. A major advantage of paired whole cell recordings is the highly precise information it can provide on the properties of synaptic transmission and plasticity that are not possible with other more crude techniques utilizing extracellular axonal stimulation. Paired whole cell recordings are often perceived as too challenging to perform. While there are challenging aspects to this technique, paired recordings can be performed by anyone trained in whole cell patch clamping provided specific hardware and methodological criteria are followed. The probability of attaining synaptically connected paired recordings significantly increases with healthy organotypic slices and stable micromanipulation allowing independent attainment of pre- and postsynaptic whole cell recordings. While CA3-CA3 pyramidal cell pairs are most widely used in the organotypic slice hippocampal preparation, this technique has also been successful in CA3-CA1 pairs and can be adapted to any neurons that are synaptically connected in the same slice preparation. In this manuscript we provide the detailed methodology and requirements for establishing this technique in any laboratory equipped for electrophysiology.

Synaptic संपर्क रिकॉर्डिंग इलेक्ट्रोड के माध्यम से एक depolarizing वर्तमान नाड़ी (20 मिसे के लिए आम तौर पर 20-50 PA) से गुजर रहा एक कार्रवाई संभावित आग को presynaptic न्यूरॉन उत्तेजक से स्पष्ट है. पोस्टअन्तर्ग्रथनी वर्तमान ट्रे?...

Watch this Scientific Journal Video about Paired Whole Cell Recordings in Organotypic Hippocampal Slices at JoVE.com

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.

Organotypic hippocampal स्लाइस में बनती पूरे सेल रिकॉर्डिंग

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct ...

Research

JoVE Journal

Neuroscience

Paired Patch Clamp Recordings from Motor-neuron and Target Skeletal Muscle in Zebrafish

मोटर न्यूरॉन से बनती पैच दबाना रिकॉर्डिंग और लक्ष्य Zebrafish में कंकाल की मांसपेशी

Larval zebrafish represent the first vertebrate model system to allow simultaneous patch clamp recording from a spinal motor-neuron and target muscle.  ...

Organotypic Hippocampal Slice Cultures

Organotypic hippocampal टुकड़ा संस्कृतियों

The hippocampus, a component of the limbic system, plays important roles in long-term memory and spatial navigation 1. Hippocampal neurons can modify the ...

Whole Cell Recording from an Organotypic Slice Preparation of Neocortex

Neocortex की एक Organotypic स्लाइस तैयारी से पूरे सेल रिकॉर्डिंग

We have been studying the expression and functional roles of voltage-gated potassium channels in pyramidal neurons from rat neocortex.  Because of the ...

Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures

बहु इलेक्ट्रोड Neuronal हिमस्खलन के Organotypic संस्कृतियों में ऐरे रिकॉर्डिंग

The cortex is spontaneously active, even in the absence of any particular input or motor output.  During development, this activity is important for the ...

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

तीव्र hippocampal स्लाइस में Synaptically पैदा Astroglial और neuronal प्रतिक्रियाएँ की दोहरी electrophysiological रिकॉर्डिंग

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

Whole-cell Patch-clamp Recordings from Morphologically- and Neurochemically-identified Hippocampal Interneurons

Morphologically- और Neurochemically पहचान Hippocampal interneurons से पूरे सेल पैच दबाना रिकॉर्डिंग

GABAergic inhibitory interneurons play a central role within neuronal circuits of the brain. Interneurons comprise a small subset of the neuronal ...

Electrophysiological and Morphological Characterization of Neuronal Microcircuits in Acute Brain Slices Using Paired Patch-Clamp Recordings

तीव्र मस्तिष्क स्लाइसें में neuronal microcircuits के electrophysiological और morphological विशेषता बनती पैच दबाना रिकॉर्डिंग का उपयोग

The combination of patch clamp recordings from two (or more) synaptically coupled neurons (paired recordings) in acute brain slice preparations with ...

Whole-cell Patch-clamp Recordings in Brain Slices

ब्रेन स्लाइस में पूरे सेल पैच दबाना रिकॉर्डिंग

Whole-cell patch-clamp recording is an electrophysiological technique that allows the study of the&#160;electrical properties of a substantial part of the ...

Organotypic Hippocampal Slice Cultures As a Model to Study Neuroprotection and Invasiveness of Tumor Cells

Organotypic हिप्पोकैम्पस एक मॉडल के रूप में Neuroprotection और ट्यूमर कोशिकाओं की इनवेसिव अध्ययन के लिए संस्कृतियों स्लाइस

In organotypic hippocampal slice cultures (OHSC), the morphological and functional characteristics of both neurons and glial cells are well preserved. ...