Name: simultaneous electroencephalography, real-time measurement of lactate concentration and optogenetic manipulation of neuronal activity in the rodent cerebral cortex
Uploaded: 2012-12-19T12:00:00
Description: Watch this Scientific Journal Video about Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral C

Исследования, финансируемого Министерством обороны (Defense Advanced Research Projects Agency, премия для молодых факультет, номер гранта N66001-09-1-2117) и NINDS (R15NS070734).

1. Хирургическая подготовка животных 1. Испытуемых Используйте мышей B6.Cg-Tg (Thy1-COP4/eYFP) 18Gfng / J трансгенной линии 9; JAX штамм # 7612) или других мышей, экспрессирующих синие светочувствительных катионов канал, Channelrhodopsin-2, в коры головного мозга нейроны. Применени...

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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+short-term+slow+wave+sleep+homeostasis+and+its+disruption+by+minocycline+in+the+laboratory+mouse.">Quantification of short-term slow wave sleep homeostasis and its disruption by minocycline in the laboratory mouse.</a> Neurosci. Lett. 490 (3), 165-169 (2011).</li><li>El Yacoubi, 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=Behavioral,+neurochemical,+and+electrophysiological+characterization+of+a+genetic+mouse+model+of+depression.">Behavioral, neurochemical, and electrophysiological characterization of a genetic mouse model of depression.</a> Proc. Natl. Acad. Sci. 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Protoc. 5 (2), 247-254 (2010).</li><li>Destexhe, A., Contreras, D., Steriade, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortically-induced+coherence+of+a+thalamic-generated+oscillation.">Cortically-induced coherence of a thalamic-generated oscillation.</a> Neuroscience. 92 (2), 427-443 (1999).</li><li>Liu, Z. W., Faraguna, U., Cirelli, C., Tononi, G., Gao, X. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+evidence+for+wake-related+increases+and+sleep-related+decreases+in+synaptic+strength+in+rodent+cortex.">Direct evidence for wake-related increases and sleep-related decreases in synaptic strength in rodent cortex.</a> J. 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Методы, представленные здесь, позволяют измерять отношения между сном и изменения в головном мозге концентрация лактата гликолитических промежуточные на шкале времени не было возможно ранее. Животные проходят спонтанные переходы между следа, NREMS и REMS. Кроме того, мы в состоянии примен...

<table border="1"><tbody><tr><td> Компонент </td><td> Компания </td><td> Номер в каталоге </td><td> Комментарии (по желанию) </td></tr><tr><td> Basi мышь канюли </td><td> Pinnacle технологии / Basi Inc </td><td> 7032 </td><td></td></tr><tr><td> Лактат биосенсоров </td><td> Pinnacle технологии </td><td> 7004 </td><td></td></tr><tr><td> Руководитель горе </td><td> Pinnacle технологии </td><td> 8402 </td><td></td></tr><tr><td> Режим сна / биосенсоров системы записи </td><td> Pinnacle технологии </td><td> 8400-K1-SL </td><td> 2 каналов ЭЭГ, 1 канал EMG, 1 и биосенсоров </td></tr><tr><td> Привязной мыши в лабораторных Набор для калибровки </td><td> Pinnacle технологии </td><td> 7000-K1-T </td><td></td></tr><tr><td> Волоконно-оптические канюли </td><td> Один пластмассы </td><td> C312G </td><td> 21 калибровочных канюли </td></tr><tr><td> Dummy Канюля </td><td> Один пластмассы </td><td> C312DC </td><td> 21 калибровочных Dummy </td></tr><tr><td> Алмазный волоконно Scribe </td><td> Thorlabs </td><td> S90W </td><td></td></tr><tr><td> Оптический разъем обжимной инструмент </td><td> Thorlabs </td><td> CT042 </td><td></td></tr><tr><td> Развилки трубы </td><td> Thorlabs </td><td> FT030 </td><td> 03,0 мм </td></tr><tr><td></td><td> Thorlabs </td><td> T10S13 </td><td> Max Dia. 0,012 </td></tr><tr><td> Stripper развилки труб </td><td> Thorlabs </td><td> FTS3 </td><td></td></tr><tr><td> Голые жесткий Покрытие многомодовых волокон </td><td> Thorlabs </td><td> BFL37-200 </td><td> 200 мкм Core, 0,37 NA </td></tr><tr><td> Провод Snips / Kevlar Ножницы </td><td> Thorlabs </td><td> T865 </td><td></td></tr><tr><td> Волоконно-оптические эпоксидные </td><td> Thorlabs </td><td> F112 </td><td></td></tr&gt; <tr><td> Инструмент волоконно Stripper </td><td> Thorlabs </td><td></td><td></td></tr><tr><td> Полировки стекла плиты </td><td> Thorlabs </td><td> CTG913 </td><td></td></tr><tr><td> Резиновые полировальные Pad </td><td> Thorlabs </td><td> NRS913 </td><td></td></tr><tr><td> Глаз Лупа </td><td> Thorlabs </td><td> JEL10 </td><td></td></tr><tr><td> Kim Wipes </td><td> Thorlabs </td><td> KW32 </td><td></td></tr><tr><td> Сжатый воздух </td><td> Thorlabs </td><td> CA3 </td><td></td></tr><tr><td> Полирование Puck </td><td> Thorlabs </td><td> D50-хх </td><td></td></tr><tr><td> Волоконно сферу инспекции </td><td> Thorlabs </td><td> CL-200 </td><td></td></tr><tr><td> Полировка фильмов </td><td> Thorlabs </td><td> LFG5P, LFG3P, LFG1P, LFG03P </td><td></td></tr><tr><td> FC / PC разъем конца </td><td> Thorlabs </tD&gt; <td> 30126G2-240 </td><td> 240 мкм Диаметр цилиндра, SS Ferrule </td></tr><tr><td> MC Стимул единицы </td><td> Многоканальных систем </td><td> STG-4002 </td><td></td></tr><tr><td> MC Стимул программного обеспечения </td><td> Многоканальных систем </td><td> MC-Стимул V 2.1.5 </td><td></td></tr><tr><td> Голубой лазер </td><td> CrystaLaser </td><td> CL473-050-0 </td><td></td></tr><tr><td> Лазерный питания </td><td> CrystaLaser </td><td> CL2005 </td><td></td></tr><tr><td> Волоконно-оптический поворотный Совместная </td><td> Дорический линзы </td><td> FRJ-v4 </td><td></td></tr><tr><td></td><td></td><td></td><td> Таблица 2. Материалов и оборудования. </td></tr></tbody></table>

simultaneous electroencephalography, real-time measurement of lactate concentration and optogenetic manipulation of neuronal activity in the rodent cerebral cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

<html> Как показано на рисунке 2, мышь оборудована для optogenetic стимуляции и лактат / ЭЭГ / ЭМГ сбора данных прошли спонтанные сон / бодрствование переходов государства, а ЭЭГ, ЭМГ и церебральной концентрации лактата контролируется постоянно. Ток при лактата датчик увеличил...

Watch this Scientific Journal Video about Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral C

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Процедура описана для управления деятельностью коры головного мозга пирамидальных нейронов optogenetically в то время как электроэнцефалограммы, электромиограммы, и церебральной концентрации лактата контролируется. Экспериментальные записи выполняются на кабельно-привязанный мышей, когда они подвергаются спонтанному сон / бодрствование циклы. Optogenetic сборки оборудования в нашей лаборатории; записывающее оборудование является коммерчески доступным.

Одновременное электроэнцефалография, в режиме реального времени измерения концентрации лактата и Optogenetic Манипуляция активности нейронов в коры головного мозга грызунов

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

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