Name: whole cell recordings from brain of adult drosophila
Uploaded: 2007-07-29T00:00:00
Description: Watch this Scientific Journal Video about Whole Cell Recordings from Brain of Adult Drosophila at JoVE.com

<p class="jove_content">This work was supported by an NIH grant NS27501 to DKOD. Additional support for this work was provided by an HHMI Professor Program Grant to DKOD.</p>

<p class="jove_step">I. Dissection of brains from adult fly</p>
<ol>
<li>Place a small drop (~100 &micro;l) of dissecting solution in center of a 35 mm Petri dish.</li>
<li>Catch adult female using aspirator. Under dissecting microscope, use two syringe needles to decapitate the fly.</li>
<li>Place head in a small drop of dissecting saline (with papain added) in a Petri dish.</li>
<li>Position head with with proboscis facing bottom of the dish.</li>
<li>Hold the cuticle covering the left compound eye with needle 1. Make a diagonal cut that exte...

<ol>
	<li>Gu, H., O Dowd, D. K., K, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholinergic+synaptic+transmission+in+Adult+Drosophila+Kenyon+cells+in+situ.">Cholinergic synaptic transmission in Adult Drosophila Kenyon cells in situ.</a> <em style='font-style: italic'>Journal of Neuroscience</em>. <b>26</b>, 265-272 (2006).</li><li>Wilson, R. I., Turner, G. C., Laurent, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+of+olfactory+representations+in+the+Drosophila+antennal+lobe.">Transformation of olfactory representations in the Drosophila antennal lobe.</a> <em style='font-style: italic'>Science</em>. <b>303</b>, 366-370 (2004).</li></ol>

<p class="jove_content">The isolated whole brain preparation we illustrate in this video allows assessment of cellular mechanisms underlying excitability and synaptic transmission in identified neurons, including those in the olfactory processing pathways, in the adult Drosophila brain (Gu and O Dowd, 2006). This approach is complementary to study of neuronal activity in the brain of intact adult Drosophila (Wilson et all 2004), in much the same way recordings from neurons in a mammalian brain slice are complementary to recordings in awake beha...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>#5 Foceps</td>
<td>Tool</td>
<td>FST by DUMONT</td>
<td>No. 11251-10 </td>
<td>Be careful, never damage the fine tips of forceps</td>
</tr>
<tr>
<td>Papain</td>
<td>Reagent</td>
<td>Worthington</td>
<td>3126</td>
<td>20 U/ml activated by 1 mM L-cysteine</td>
</tr>
<tr>
<td>Dissecting microscope</td>
<td>SMZ-2B Model:C-PS</td>
<td>Nikon</td>
<td>&nbsp;</td>
<td>Equipped with gooseneck fiber optic lights positioned a low angle</td>
</tr>
<tr>
<td>Needle & Syringe </td>
<td>&nbsp;</td>
<td>PrecisionGlide&#174;?, Becton Dickinson & Co </td>
<td>305175 & 309602</td>
<td>Cheap and durable</td>
</tr>
<tr>
<td>Upright microscope</td>
<td>Axioskop2 FS plus</td>
<td>Zeiss</td>
<td>&nbsp;</td>
<td>Equipped with a fixed stage, 40X water immersion objective, Nomarski optics, and fluorescent lamp</td>
</tr>
<tr>
<td>Patch clamp amplifier</td>
<td>200 B</td>
<td>Molecular device</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Digidata  D-A converter </td>
<td>1322A</td>
<td>Molecular device</td>
<td>&nbsp;</td>
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whole cell recordings from brain of adult drosophila

In this video, we demonstrate the procedure for isolating whole brains from adult Drosophila in preparation for recording from single neurons. We begin by describing the dissecting solution and capture of the adult females used in our studies. The procedure for removing the whole brain intact, including both optic lobes, is illustrated. Dissection of the overlying trachea is also shown.  The isolated brain is not only small but needs special care in handling at this stage to prevent damage to the neurons, many of which are close to the outer surface of the tissue. We show how a special holder we developed is used to stabilize the brain in the recording chamber. A standard electrophysiology set up is used for recording from single neurons or pairs of neurons. A fluorescent image, viewed through the recording microscope, from a GAL4 line driving GFP expression (GH146) illustrates how projection neurons (PNs) are identified in the live brain. A high power Nomarski image shows a view of a single neuron that is being targeted for whole cell recording. When the brain is successfully removed without damage, the majority of the neurons are spontaneously active, firing action potentials and/or exhibiting spontaneous synaptic input. This in situ preparation, in which whole cell recording of identified neurons in the whole brain can be combined with genetic and pharmacological manipulations, is a useful model for exploring cellular physiology and plasticity in the adult CNS.

Watch this Scientific Journal Video about Whole Cell Recordings from Brain of Adult Drosophila at JoVE.com

Whole Cell Recordings from Brain of Adult Drosophila

<p class="jove_content">This video demonstrates the procedure for isolating whole brains from adult Drosophila in preparation for recording from single neurons using standard whole cell technology.  It includes images of GFP labeled cells and neurons viewed during recording.</p>

In this video, we demonstrate the procedure for isolating whole brains from adult Drosophila in preparation for recording from single neurons. We begin by ...

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