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Author (up) Bucher, D.; Buchner, E. file  url
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  Title Stimulating PACalpha increases miniature excitatory junction potential frequency at the Drosophila neuromuscular junction Type Journal Article
  Year 2009 Publication Journal of Neurogenetics Abbreviated Journal J Neurogenet  
  Volume 23 Issue 1-2 Pages 220-224  
  Keywords Adenylyl Cyclases/*physiology; Animals; Cyclic AMP/physiology; Drosophila/*physiology; Enzyme Activation/radiation effects; Excitatory Postsynaptic Potentials/physiology; Light Signal Transduction/physiology; Miniature Postsynaptic Potentials/physiology; Motor Neurons/enzymology; Neuromuscular Junction/*physiology; Photic Stimulation/methods; Synapses/enzymology/physiology  
  Abstract Photoactivated adenylate cyclase alpha (PACalpha) is a light-activated adenylate cyclase that was originally cloned from the eye spot of the protozoan Euglena gracilis. PACalpha has been shown to rapidly increase intracellular cyclic adenosine monophosphate (cAMP) in vivo in Xenopus oocytes and HEK293 cells, increase the spike width in Aplysia sensory neurons, and modify behavior in Drosophila. Using the GAL4 UAS system, we heterologously expressed PACalpha in motorneurons and quantified the effects of its activation at the neuromuscular junction of the Drosophila third instar wandering larva, a well-characterized model synapse. By recording from body-wall muscle 6, we show that the presynaptic activation of PACalpha with blue light significantly increased miniature excitatory junction potential (mEJP) frequency in the presence of calcium with a delay of about 1 minute. Similar effects have been observed in previous studies that utilized adenylate cyclase agonists (Forskolin) or membrane-permeable cAMP analogs [dibutyryl cAMP and 4-chlorophenylthio-(CPT)-cAMP] to increase presynaptic cAMP concentrations. PACalpha's efficacy in combination with its specificity make it an invaluable tool for the rapid regulation of cAMP in vivo and for investigating the mechanisms by which cAMP can modulate synaptic transmission and neuronal plasticity in Drosophila.  
  Call Number Serial 1255  
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Author (up) Zhang, B.; Stewart, B. file  url
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  Title Electrophysiological recording from Drosophila larval body-wall muscles Type Journal Article
  Year 2010 Publication Cold Spring Harbor Protocols Abbreviated Journal Cold Spring Harb Protoc  
  Volume 2010 Issue 9 Pages pdb.prot5487  
  Keywords Animals; Cytosol/physiology; Drosophila/*physiology; Electrophysiology/instrumentation/*methods; Muscles/physiology; Neuromuscular Junction/physiology  
  Abstract The Drosophila larval neuromuscular junction (NMJ) shares many structural and functional similarities to synapses in other animals, including humans. These include the basic feature of synaptic transmission, as well as the molecular mechanisms regulating the synaptic vesicle cycle. Because of its large size, easy accessibility, and the well-characterized genetics, the fly NMJ remains an excellent model system for dissecting the cellular and molecular mechanisms of synaptic transmission. This protocol describes the steps for performing intracellular recording from fly larval body-wall muscles and explains how to record and analyze spontaneous and evoked synaptic potentials. Methods used include larval dissection (“filleting”), identification of muscle fibers and their innervating nerves, the use of a micromanipulator and microelectrode in penetrating the muscle membrane, and nerve stimulation to evoke synaptic potentials.  
  Call Number Serial 1323  
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