Supported by APA to RJH, NHMRC program grant 400121 to SP, and NMHRC fellowship 1005050 to SP. CAR acknowledges the support of the ARC (FT0990628) and the DOWD fellowship scheme. The Florey Institute of Neuroscience and Mental Health is supported by Victorian State Government infrastructure funds.

All experiments on mice were approved by the Florey Institute animal ethics committee.
1. Setup for Cutting Brain Slices
<ol>
	<li>Prepare a cutting solution comprised of (mM) 125 Choline-Cl, 2.5 KCl, 0.4 CaCl2, 6 MgCl2, 1.25 NaH2PO4, 26 NaHCO3, 20 D-glucose saturated with carbogen gas (95% O2&#8211;5% CO2) and an artificial cerebrospinal fluid (aCSF) recording solution comprised of (mM) 125 NaCl, 2.5 KCl, 2&#160;Ca...

<ol>
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A robust method to generate CA1 &#947; oscillations in acute brain slices is described. The oscillations generated arise from a local circuit enabling a better opportunity for controlling and understanding the neurophysiological basis of network oscillations12. AMPA receptors, GABAA receptors, Ih and T-type Ca2+ channels are all required for &#947; oscillations in this model. While the local CA1 oscillations described here can be robustly generated this is dependent on ensuring...

Brain network oscillations occur within distinct frequency bands that correlate to behavioral states. In rodents, hippocampal &#952; oscillations (5 - 10 Hz) are observed during exploratory behaviors1,2, while &#947; oscillations (20 - 80 Hz) associate with various cognitive processes, including perception and attention3,4. Synchronous &#947; network activity is also implicated in the pathology of disorders such as epilepsy and schizophrenia5,6. For example, &#947; oscillations are thought to correspond to areas of cortical epileptic foci5,7,8 and could be used as markers of pharmacosensitivity or resistance, two important areas of investigation in epilepsy research9.
The hippocampal brain slice is a model that has been widely used to investigate network activity10-12. Various protocols have been developed to generate &#947; oscillations in brain slices that typically involve pharmacological modulation such as low Mg2+, 4-aminopyridine (4AP), bicuculline, and kainic acid12-17. Shortcomings of pharmacologically triggered oscillations are that they occur randomly after drug application and are not reliably generated or stable over time. Electrically triggered &#947; oscillations overcome many of these problems and also have the advantage of being temporally locked to the stimulating event allowing for episodic recording and analysis. Here a protocol is described for generating CA1 &#947; oscillations by delivering a tetanic stimulation to the stratum oriens in the hippocampal slice.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>4-(N-Ethyl-N-phenylamino)-1,2- dimethyl-6-(methylamino) pyrimidinium chloride (ZD7288)</td>
			<td>Sigma-Aldrich</td>
			<td>Z3777</td>
			<td></td>
		</tr>
		<tr>
			<td>Biuculline</td>
			<td>Sigma-Aldrich</td>
			<td>14340</td>
			<td></td>
		</tr>
		<tr>
			<td>6-cyano-7-nitroquinoxa- line-2,3-dione (CNQX)</td>
			<td>Sigma-Aldrich</td>
			<td>C127</td>
			<td></td>
		</tr>
		<tr>
			<td>Nickel</td>
			<td>Sigma-Aldrich</td>
			<td>266965</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbamazepine</td>
			<td>Sigma-Aldrich</td>
			<td>C4024</td>
			<td></td>
		</tr>
		<tr>
			<td>(2R)-amino-5-phosphonopentano-ate (APV)</td>
			<td>Tocris Bioscience</td>
			<td>0105</td>
			<td></td>
		</tr>
		<tr>
			<td>Retigabine</td>
			<td>ChemPacific</td>
			<td>150812-12-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Choline-Cl</td>
			<td>Sigma Aldrich</td>
			<td>C1879-5KG</td>
			<td></td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma Aldrich</td>
			<td>P9333-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Sigma Aldrich</td>
			<td>S9638-250G</td>
			<td></td>
		</tr>
		<tr>
			<td>NaHCO3</td>
			<td>Sigma Aldrich</td>
			<td>S6297-250G</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma Aldrich</td>
			<td>S7653-5KG</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>Sigma Aldrich</td>
			<td>G8270-1KG</td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2.2H2O</td>
			<td>Sigma Aldrich</td>
			<td>223506-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>MgCl2.6H2O</td>
			<td>Sigma Aldrich</td>
			<td>M2670-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode glass</td>
			<td>Harvard Apparatus&#160;</td>
			<td>GC150F-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Concentric bipolar stimulating metal electrode&#160;</td>
			<td>FHC</td>
			<td>CBBPF75</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital Isolator</td>
			<td>Getting Instruments</td>
			<td>Model BJN8-9V1&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Model 1800 amplifier</td>
			<td>A-M systems</td>
			<td>Model 1800 amplifier</td>
			<td></td>
		</tr>
		<tr>
			<td>Digitizer</td>
			<td>National Intruments</td>
			<td>NI USB-6211</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibrotome</td>
			<td>Leica</td>
			<td>VT1200s</td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of local ca1 &#947; oscillations by tetanic stimulation

Neuronal network oscillations are important features of brain activity in health and disease and can be modulated by a range of clinically used drugs. A protocol is provided to generate a model for studying CA1 &#947; oscillations (20 - 80 Hz). These &#947; oscillations are stable for at least 30 min&#160;and depend upon excitatory and inhibitory synaptic activity in addition to activation of pacemaker currents. Tetanically stimulated oscillations have a number of reproducible and easily quantifiable characteristics including spike count, oscillation duration, latency and frequency that report upon the network state. The advantages of the electrically stimulated oscillations include stability, reproducibility and episodic acquisition enabling robust characterization of network function. This model of CA1 &#947; oscillations can be used to study cellular mechanisms and to systematically investigate how neuronal network activity is altered in disease and by drugs. Disease state pharmacology can be readily incorporated by the use of brain slices from genetically modified or interventional animal models to enable selection of drugs that specifically target disease mechanisms.

Tetanic stimulation of the stratum oriens generated robust and reproducible &#947; oscillations (35.4 &#177; 2.2&#160;Hz), see Figure 3B. To demonstrate that the oscillations were generated within the local CA1 network the inputs from CA3 were severed by cutting the slice in the CA2 region using a bent 32&#160;G needle. The oscillation properties in the cut slices did not differ from the uncut slices (p = 0.85; cut slices 6.16 &#177; 1.1 spikes, n = 6; uncut slices 5.89 &#177; 0.8 spikes, n = 6), indicat...

Watch this Scientific Journal Video about Generation of Local CA1 Î³ Oscillations by Tetanic Stimulation at JoVE.com

Generation of Local CA1 &amp;#947; Oscillations by Tetanic Stimulation

Oscillations are fundamental network properties and are modulated by disease and drugs. Studying brain-slice oscillations allows characterization of isolated networks under controlled conditions. Protocols are provided for the preparation of acute brain slices for evoking CA1 &#947; oscillations.

Generation of Local CA1 &#947; Oscillations by Tetanic Stimulation

Neuronal network oscillations are important features of brain activity in health and disease and can be modulated by a range of clinically used drugs. A ...

generation-of-local-ca1-oscillations-by-tetanic-stimulation

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Neuroscience

8.9K Aufrufe.  University of Melbourne.  Schwingungen sind grundlegende Netzeigenschaften und sind von Krankheiten und Arzneimittel moduliert. Studieren Gehirn-slice Schwingungen ermöglicht Charakterisierung der isolierten Netze unter kontrollierten Bedingungen. Protokolle sind für die Herstellung von akuten Gehirnscheiben zum Hervorrufen CA1 γ Schwingungen vorgesehen. 