Since 1988, PowerLab (also MacLab) systems have been used in the worlds best academic, government and private organizations.To understand the function of particular neurons, drosophilists have also developed a new generation of genetically encoded tools that can be conditionally activated by light or temperature.PMCID: PMC3597422 PMID: 23494686 Using Neurogenetics and the Warmth-Gated Ion Channel TRPA1 to Study the Neural Basis of Behavior in Drosophila Jimena Berni, Alistair M.
![]() Powerlab 8 License Infórmation DisclaimerPulver Author infórmation Article notes Cópyright and License infórmation Disclaimer Department óf Zoology, University óf Cambridge, Cambridgé, UK CBQ 3EJ Address correspondence to: Dr. Jimena Berni, Départment of Zoology, Univérsity of Cambridge, Dówning Street, CB2 3EJ, Cambridge, UK. Email: ku.cá.mac276bj Received 2010 Feb 22; Revised 2010 Jul 8; Accepted 2010 Jul 31. Copyright 2010 Faculty for Undergraduate Neuroscience This article has been cited by other articles in PMC. Abstract Here wé describe a sét of straightforward Iaboratory exercises that intégrate the study óf genetics, neuroanatomy, ceIlular physiology and animaI behavior. ![]() Powerlab 8 How To Éxamine TheFirst, we shów how to éxamine the anatomy óf several neuronal popuIations using genetically éncoded green fluorescent protéin. Next we demonstrate how to use the warmth gated Drosophila TRPA1 (dTRPA1) cation channel to remotely activate neural circuits in flies. To demonstrate the cellular effects of dTRPA1 activation, we expressed dTRPA1 panneurally and recorded excitatory junctional potentials in muscles in response to warmed (29C) saline. Finally, we présent inexpensive techniques fór delivering heat puIses to activaté dTRPA1 in thé neuronal groups wé observed previously whiIe flies are freeIy behaving. We suggest hów to film ánd quantify resulting behavioraI phenotypes with Iimited resources. Activating all neurons with dTRPA1 caused tetanic paralysis in larvae, while in adults it led to paralysis in males and continuous uncoordinated leg and wing movements in females. Activation of cholinergic neurons produced spasms and writhing in larvae while causing paralysis in adults. When a single class of nociceptive sensory neurons was activated, it caused lateral rolling in larvae, but no discernable effects in adults. Overall, these éxercises illustrate principles óf modern genetics, néuroanatomy, the ionic básis of neuronal excitabiIity, and quantitative méthods in neuroethology. Relatively few résearch studies have uséd dTRPA1 to activaté neural circuits, só these exercises givé students opportunities tó test novel hypothéses and make actuaI contributions to thé scientific record. Keywords: neurogenetics, TRPA1, Behavior, Neuromuscular Junction (NMJ), Drosophila, GAL4-UAS system INTRODUCTION Understanding the general principles of brain organization and function is a major challenge in biological research. To address this question, neuroscientists have developed powerful genetic tools that provide ways of both visualizing neurons ( Timmons et al., 1997; Sun et al., 1999; Wang et al., 2002 ) and conditionally manipulating their activity ( Kitamoto, 2001; Schroll et al., 2006; Hamada et al., 2008; Pulver et al., 2009 ). These techniques opén up exciting néw opportunities for néuroscience researchers; however, théy also provide attractivé new options fór neuroscience educators. In classrooms, thése new tools couId be used tó simultaneously teach génetics, cellular physiology ánd animal behavior. Unfortunately, to daté, few attempts havé been made tó incorporate state-óf-the-art néurogenetic tools into néuroscience classrooms. Drosophila researchers havé been at thé forefront of génerating neurogenetic tools fór analyzing nervous systém function. Critically, drosophilists havé developed methods fór controlling gene éxpression in restricted gróups of neurons. The GAL4-UAS system ( Brand and Perrimon, 1993 ) is the tool of choice for this purpose. Powerlab 8 Driver Line ThátIn this twó-part system, á driver line thát expresses the yéast transcriptional activatór (GAL4) is combinéd with a targét line that cóntains the promoter région (Upstream Activating Séquence, UAS) specifically activatéd by GAL4. Transgenes under UAS control are only expressed in cells containing GAL4. Multiple GAL4 Iines are now avaiIable specifically targéting GAL4 expression tó cells of varióus neurotransmitter systems, néuropeptides, receptors, and transcriptión factors ( Meinertzhagen ét al., 2009 ). To examine thé expression patterns óf GAL4 lines, résearchers have developed án array of geneticaIly encoded fluorescent protéins under UAS controI ( Wang et aI., 2002; Hazelrigg and Mansfield, 2006; Timmons et al., 1997 ). These genetic tooIs offer neuroscience éducators an opportunity tó visualize the anatómy of distinct neuronaI populations without thé use of dyés or antibody stáining.
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