Name: harvesting and preparing <em>drosophila</em> embryos for electrophysiological recording and other procedures
Uploaded: 2009-05-20T00:00:00
Description: Watch this Scientific Journal Video about Harvesting and Preparing Drosophila Embryos for Electrophysiological Recording and Other Procedures at JoVE.com

<p class="jove_content">K.B. is supported by NIH grant GM54544.</p>

<p class="jove_title">Part 1: Equipment and Supplies</p>
<ol>
<li>A good dissection microscope is required for embryo dissections; 40X magnification is suggested, with 25X eye-pieces to maximally increase magnification.</li>
<li>Fine forceps (number 5) are required for manual selection of embryos and devitellinization of embryos.</li>
<li>Equipment to make and modify fine glass needles is required.  Pulled-glass needles are required for the dissection. We prefer solid glass for dissection (last longer), but standard thick-walled glass tubing (o...

<p class="jove_content">Precise staging of embryos is critical due to the rapid maturation of functional properties over the time course of just several hours. Several issues complicate this staging. First, most researchers use timed egg lays to stage embryos, but the egg laying time can vary enormously from animal to animal in different conditions (Broadie et al. 1992). In particular, females on a limited diet tend to retain fertilized eggs for prolonged periods prior to laying. It is therefore critical to &ldquo;clear&rdquo; females by feedin...

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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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<td>Embryo Collection Cages</td>
<td>&nbsp;</td>
<td>Genesee Scientific (www.flystuff.com)</td>
<td>59-100 (for 60 mm dish; other sizes available)</td>
<td>Cages can also be home made using punctured tri-pour beakers, as shown in video</td>
</tr>
<tr>
<td>Sylgard 184</td>
<td>&nbsp;</td>
<td>Dow Corning</td>
<td>Available from various companies</td>
<td>Surgical glue adheres better to sylgard-coated coverslips</td>
</tr>
<tr>
<td>Fine glass tubing, outer diameter 1.0-1.5 mm</td>
<td>&nbsp;</td>
<td>various</td>
<td>&nbsp;</td>
<td>For pulling into fine glass needles for dissection and tubes for glue delivery</td>
</tr>
<tr>
<td>Plastic tubing, to attach to glass pipette for mouth suction and glue application</td>
<td>&nbsp;</td>
<td>Tygon</td>
<td>&nbsp;</td>
<td>Tubing inner diameter needs to match glass outer diameter.</td>
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harvesting and preparing <em>drosophila</em> embryos for electrophysiological recording and other procedures

<em>Drosophila</em> is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite isolated from each other, with largely independent histories and scientific communities. However, the interface between these usually disparate fields is the developmental programs underlying acquisition of functional electrical signaling properties and differentiation of functional chemical synapses during the final phases of neural circuit formation. This interface is a critically important area for investigation. In <em>Drosophila</em>, these phases of functional development occur during a period of &lt;8 hours (at 25&deg;C) during the last third of embryogenesis. This late developmental period was long considered intractable to investigation owing to the deposition of a tough, impermeable epidermal cuticle. A breakthrough advance was the application of water-polymerizing surgical glue that can be locally applied to the cuticle to enable controlled dissection of late-stage embryos. With a dorsal longitudinal incision, the embryo can be laid flat, exposing the ventral nerve cord and body wall musculature to experimental investigation. This system has been heavily used to isolate and characterize genetic mutants that impair embryonic synapse formation, and thus reveal the molecular mechanisms governing the specification and differentiation of synapse connections and functional synaptic signaling properties.

Watch this Scientific Journal Video about Harvesting and Preparing Drosophila Embryos for Electrophysiological Recording and Other Procedures at JoVE.com

Harvesting and Preparing Drosophila Embryos for Electrophysiological Recording and Other Procedures

<p class="jove_content">This technique exposes the Drosophila embryonic neuromusculature for immunohistochemistry or electrophysiological recording.  It is useful for studying early events in neuromuscular development or performing electrophysiology in mutants that cannot hatch.</p>

Harvesting and Preparing <em>Drosophila</em> Embryos for Electrophysiological Recording and Other Procedures

Drosophila is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite ...

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