K.B. is supported by NIH grant GM54544.

Part 1: Equipment and Supplies
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<li>Electrophysiological recording from Drosophila embryos first requires proficiency in embryonic dissection techniques, which are described in another JoVE video.</li>
<li>Electrophysiological recording from Drosophila embryos utilizes standard patch clamp recording configurations. Patch clamp recording equipment and software suitable for many other preparations is also suitable for recording from Drosophila embryos. Because Drosophila embryos ...

Electrophysiological recording from Drosophila embryos requires manual manipulation and dissection. The health of the preparation, and consequent quality of the recordings, depends on one being able to quickly and neatly prepare the fragile embryonic tissues for recording, and then execute the experiment. Experimenters should be proficient in both embryonic dissections and patch clamp electrophysiology before trying to tackle both at once.
Recordings can be done in one of several var...

electrophysiological recording in the drosophila embryo

Drosophila 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 Drosophila, 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. Whole-cell patch-clamp techniques can then be employed to record from individually-identifiable neurons and somatic muscles. These recording configurations have been used to track the appearance and maturation of ionic currents and action potential propagation in both neurons and muscles. Genetic mutants affecting these electrical properties have been characterized to reveal the molecular composition of ion channels and associated signaling complexes, and to begin exploration of the molecular mechanisms of functional differentiation. A particular focus has been the assembly of synaptic connections, both in the central nervous system and periphery. The glutamatergic neuromuscular junction (NMJ) is most accessible to a combination of optical imaging and electrophysiological recording. A glass suction electrode is used to stimulate the peripheral nerve, with excitatory junction current (EJC) recordings made in the voltage-clamped muscle. This recording configuration has been used to chart the functional differentiation of the synapse, and track the appearance and maturation of presynaptic glutamate release properties. In addition, postsynaptic properties can be assayed independently via iontophoretic or pressure application of glutamate directly to the muscle surface, to measure the appearance and maturation of the glutamate receptor fields. Thus, both pre- and postsynaptic elements can be monitored separately or in combination during embryonic synaptogenesis. 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 Electrophysiological Recording in the Drosophila Embryo at JoVE.com

Electrophysiological Recording in the Drosophila Embryo

Electrophysiological recordings from Drosophila embryos allow analyses of developing muscle and neuron electrical properties, as well as characterization of functional synaptogenesis at the glutamatergic neuromuscular junction and central cholinergic and GABAergic synapses.

Electrophysiological Recording in the Drosophila Embryo

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

electrophysiological-recording-in-the-drosophila-embryo

Research

JoVE Journal

Biology

11.3K visualizações.  University of Illinois. Gravações eletrofisiológicas da Drosophila Embriões permitem análises de músculo em desenvolvimento e as propriedades elétricas do neurônio, bem como caracterização de sinaptogênese funcionais da junção neuromuscular e glutamatérgica colinérgica central e sinapses GABAérgica.