Authorea

Walton Jones refs almost 9 years ago

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Sleep and locomotor behavior of flies were measured using Drosophila Activity Monitoring system (Trikinetics). For sleep analysis, 3-4 day-old female flies were placed into 65 mm X 5mm glass tube individually with one end filled with 2\% agar/ 5\% sucrose food and the other end capped by cotton ball. Activity was monitored as every bim break in 1 minute bin and 5 minutes and more inactivity were counted as sleep \cite{10710313}. \cite{Shaw:2000ui}. After one day of habituation in the incubator (25$\,^{\circ}\mathrm{C}$, 60\% humidity), sleep during two days of 12hr:12hr light-dark cycle and following two days of dark period was analysed using Counting Macro \cite{21041393}. For circadian locomotor analysis, 1-3 day-old male flies were used. Activity was measured every 30 min bin and analysed using ClockLab analysis software (Actimetrics) and Counting Macro\cite{21041392}. Significance level of the $\chi$\textsuperscript{2} periodogram was set to $\alpha$ = 0.05. Flies with a power of significance (P-S) $\geq$10 were considered as rhythmic.

%% http://bibdesk.sourceforge.net/ %% Created for Walton Jones at 2015-07-18 21:09:44 21:20:08 +0900 %% Saved with string encoding Unicode (UTF-8) @article{Young:2010jq, Affiliation = {Institute for Adaptive and Neural Computation, University of Edinburgh, Edinburgh, UK. [email protected]}, Author = {Young, J M and Armstrong, J D}, Date-Added = {2015-07-18 12:19:25 +0000}, Date-Modified = {2015-07-18 12:19:25 +0000}, Doi = {10.1002/cne.22284}, File = {{Young2010jq.pdf:/Users/walton/Google Drive/Papers/2010/Y/Young2010jq.pdf:application/pdf}}, Journal = {J Comp Neurol}, Keywords = {fly, behavior}, Language = {English}, Month = may, Number = {9}, Pages = {1500--1524}, Pmid = {20187142}, Rating = {0}, Title = {{Structure of the adult central complex in Drosophila: organization of distinct neuronal subsets.}}, Volume = {518}, Year = {2010}, Abstract = {The central complex (CX) is a defined set of neuropils located on the midline of the protocerebrum in several arthropods and has been implicated in a number of behaviors. To investigate the function of the CX further it is imperative to know the neuroarchitecture of this structure and to ensure all known neuron types conform to a common nomenclature system. Several types of CX neuron have been identified but it is not known if these exist singly or as components of isomorphic sets. We used an enhancer trap approach to study the adult structure, connectivity, and polarity of CX neurons in Drosophila. We observed several isomorphic sets of small-field neurons including pontine and fb-eb neurons, and also isomorphic sets of large-field neurons including R neurons and F neurons. We found that several types of large-field F neurons existed in isomorphic sets of approximately eight (four per hemisphere) and found evidence for small-field neuron types existing as isomorphic sets of 16. Small-field neurons were observed in clearly organized layers. This study provides a novel insight into CX structure and connectivity and provides a set of characterized enhancer trap lines that will be valuable for future study.}, Bdsk-File-1 = {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}, Bdsk-Url-1 = {http://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=20187142&retmode=ref&cmd=prlinks}, Bdsk-Url-2 = {http://dx.doi.org/10.1002/cne.22284}} @article{Kitamoto:2001ue, Affiliation = {Division of Neurosciences, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, Duarte, California 91010, USA. [email protected]}, Author = {Kitamoto, T}, Date-Added = {2015-07-18 12:17:04 +0000}, Date-Modified = {2015-07-18 12:17:21 +0000}, Doi = {10.1002/neu.1018}, File = {{Kitamoto2001ue.pdf:/Users/walton/Google Drive/Papers/2001/K/Kitamoto2001ue.pdf:application/pdf}}, Journal = {J Neurobiol}, Language = {eng}, Month = may, Number = {2}, Pages = {81--92}, Pmid = {11291099}, Rating = {0}, Title = {{Conditional modification of behavior in Drosophila by targeted expression of a temperature-sensitive shibire allele in defined neurons}}, Volume = {47}, Year = {2001}, Abstract = {Behavior is a manifestation of temporally and spatially defined neuronal activities. To understand how behavior is controlled by the nervous system, it is important to identify the neuronal substrates responsible for these activities, and to elucidate how they are integrated into a functional circuit. I introduce a novel and general method to conditionally perturb anatomically defined neurons in intact Drosophila. In this method, a temperature-sensitive allele of shibire (shi(ts1)) is overexpressed in neuronal subsets using the GAL4/UAS system. Because the shi gene product is essential for synaptic vesicle recycling, and shi(ts1) is semidominant, a simple temperature shift should lead to fast and reversible effects on synaptic transmission of shi(ts1) expressing neurons. When shi(ts1) expression was directed to cholinergic neurons, adult flies showed a dramatic response to the restrictive temperature, becoming motionless within 2 min at 30 degrees C. This temperature-induced paralysis was reversible. After being shifted back to the permissive temperature, they readily regained their activity and started to walk in 1 min. When shi(ts1) was expressed in photoreceptor cells, adults and larvae exhibited temperature-dependent blindness. These observations show that the GAL4/UAS system can be used to express shi(ts1) in a specific subset of neurons to cause temperature-dependent changes in behavior. Because this method allows perturbation of the neuronal activities rapidly and reversibly in a spatially and temporally restricted manner, it will be useful to study the functional significance of particular neuronal subsets in the behavior of intact animals.}, Annote = {Temperature-sensitive shibire}, Bdsk-File-1 = {YnBsaXN0MDDUAQIDBAUGJCVYJHZlcnNpb25YJG9iamVjdHNZJGFyY2hpdmVyVCR0b3ASAAGGoKgHCBMUFRYaIVUkbnVsbNMJCgsMDxJXTlMua2V5c1pOUy5vYmplY3RzViRjbGFzc6INDoACgAOiEBGABIAFgAdccmVsYXRpdmVQYXRoWWFsaWFzRGF0YV8QPC4uLy4uLy4uLy4uLy4uL0dvb2dsZSBEcml2ZS9QYXBlcnMvMjAwMS9LL0tpdGFtb3RvMjAwMXVlLnBkZtIXCxgZV05TLmRhdGFPEQG4AAAAAAG4AAIAAAxNYWNpbnRvc2ggSEQAAAAAAAAAAAAAAAAAAADQZvGzSCsAAABHtIUSS2l0YW1vdG8yMDAxdWUucGRmAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAEfuMcXAawUAAAAAAAAAAAAFAAUAAAkgAAAAAAAAAAAAAAAAAAAAAUsAABAACAAA0GZzIwAAABEACAAAxb/sdQAAAAEAGABHtIUAR7PGAEezvwBHszoAEOOeABDjhQACAE1NYWNpbnRvc2ggSEQ6VXNlcnM6AHdhbHRvbjoAR29vZ2xlIERyaXZlOgBQYXBlcnM6ADIwMDE6AEs6AEtpdGFtb3RvMjAwMXVlLnBkZgAADgAmABIASwBpAHQAYQBtAG8AdABvADIAMAAwADEAdQBlAC4AcABkAGYADwAaAAwATQBhAGMAaQBuAHQAbwBzAGgAIABIAEQAEgA6VXNlcnMvd2FsdG9uL0dvb2dsZSBEcml2ZS9QYXBlcnMvMjAwMS9LL0tpdGFtb3RvMjAwMXVlLnBkZgATAAEvAAAVAAIADf//AACABtIbHB0eWiRjbGFzc25hbWVYJGNsYXNzZXNdTlNNdXRhYmxlRGF0YaMdHyBWTlNEYXRhWE5TT2JqZWN00hscIiNcTlNEaWN0aW9uYXJ5oiIgXxAPTlNLZXllZEFyY2hpdmVy0SYnVHJvb3SAAQAIABEAGgAjAC0AMgA3AEAARgBNAFUAYABnAGoAbABuAHEAcwB1AHcAhACOAM0A0gDaApYCmAKdAqgCsQK/AsMCygLTAtgC5QLoAvoC/QMCAAAAAAAAAgEAAAAAAAAAKAAAAAAAAAAAAAAAAAAAAwQ=}, Bdsk-Url-1 = {http://www3.interscience.wiley.com/journal/78504228/abstract?CRETRY=1&SRETRY=0}} @article{Shaw:2000ui, Affiliation = {The Neurosciences Institute, 10640 John Jay Hopkins Drive, San Diego, CA 92121, USA.}, Author = {Shaw, P J and Cirelli, C and Greenspan, Ralph J and Tononi, G}, Date-Added = {2015-07-18 12:16:05 +0000}, Date-Modified = {2015-07-18 12:16:05 +0000}, File = {{Shaw2000ui.pdf:/Users/walton/Google Drive/Papers/2000/S/Shaw2000ui.pdf:application/pdf}}, Journal = {Science}, Language = {eng}, Month = mar, Number = {5459}, Pages = {1834--1837}, Pmid = {10710313}, Rating = {0}, Title = {{Correlates of sleep and waking in Drosophila melanogaster}}, Volume = {287}, Year = {2000}, Abstract = {Drosophila exhibits a circadian rest-activity cycle, but it is not known whether fly rest constitutes sleep or is mere inactivity. It is shown here that, like mammalian sleep, rest in Drosophila is characterized by an increased arousal threshold and is homeostatically regulated independently of the circadian clock. As in mammals, rest is abundant in young flies, is reduced in older flies, and is modulated by stimulants and hypnotics. Several molecular markers modulated by sleep and waking in mammals are modulated by rest and activity in Drosophila, including cytochrome oxidase C, the endoplasmic reticulum chaperone protein BiP, and enzymes implicated in the catabolism of monoamines. Flies lacking one such enzyme, arylalkylamine N-acetyltransferase, show increased rest after rest deprivation. These results implicate the catabolism of monoamines in the regulation of sleep and waking in the fly and suggest that Drosophila may serve as a model system for the genetic dissection of sleep.}, Bdsk-File-1 = {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}, Bdsk-Url-1 = {http://www.sciencemag.org/cgi/content/full/287/5459/1834}} @article{Iniguez:2013ib, Affiliation = {Department of Developmental and Cell Biology and Department of Anatomy and Neurobiology, University of California, Irvine, California.}, Author = {Iniguez, Jorge and Schutte, Soleil S and O'Dowd, Diane K}, Date-Added = {2015-07-18 12:14:51 +0000}, Date-Modified = {2015-07-18 12:14:51 +0000}, Doi = {10.1152/jn.00368.2013}, File = {{Iniguez2013ib.pdf:/Users/walton/Google Drive/Papers/2013/I/Iniguez2013ib.pdf:application/pdf}}, Journal = {J Neurophysiol}, Language = {English}, Month = oct, Number = {7}, Pages = {1490--1496}, Pmid = {23864373}, Rating = {0}, Title = {{Cav3-type $\alpha$1T calcium channels mediate transient calcium currents that regulate repetitive firing in Drosophila antennal lobe PNs.}}, Volume = {110}, Year = {2013}, Abstract = {Projection neurons (PNs), located in the antennal lobe region of the insect brain, play a key role in processing olfactory information. To explore how activity is regulated at the level of single PNs within this central circuit we have recorded from these neurons in adult Drosophila melanogaster brains. Our previous study demonstrated that PNs express voltage-gated calcium currents with a transient and sustained component. We found that the sustained component is mediated by cac gene-encoded Cav2-type channels involved in regulating action potential-independent release of neurotransmitter at excitatory cholinergic synapses. The function of the transient calcium current and the gene encoding the underlying channels, however, were unknown. Here we report that the transient current blocked by prepulse inactivation is sensitive to amiloride, a vertebrate Cav3-type channel blocker. In addition PN-specific RNAi knockdown of $\alpha$1T, the Drosophila Cav3-type gene, caused a dramatic reduction in the transient current without altering the sustained component. These data demonstrate that the $\alpha$1T gene encodes voltage-gated calcium channels underlying the amiloride-sensitive transient current. Alterations in evoked firing and spontaneous burst firing in the $\alpha$1T knockdowns demonstrate that the Cav3-type calcium channels are important in regulating excitability in adult PNs.}, Bdsk-File-1 = {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}, Bdsk-Url-1 = {http://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=23864373&retmode=ref&cmd=prlinks}, Bdsk-Url-2 = {http://dx.doi.org/10.1152/jn.00368.2013}} @article{alphen:2013aa, Author = {van Alphen, B. and Yap, M. H. W. and Kirszenblat, L. and Kottler, B. and van Swinderen, B.}, Date-Added = {2015-07-17 22:46:47 +0000},

Volume = {2010}, Year = {2010}} @article{10710313, Author = {Shaw, PJ and Cirelli, C and Greenspan, RJ and Tononi, G}, Authors = {Shaw, PJ and Cirelli, C and Greenspan, RJ and Tononi, G}, Date = {2000 Mar 10}, Journal = {Science}, Month = {Mar}, Pages = {1834-7}, Pubmed_Id = {10710313}, Source = {Science}, Title = {{Correlates of sleep and waking in Drosophila melanogaster.}}, Volume = {287}, Year = {2000}} @article{21041392, Author = {Pfeiffenberger, C and Lear, BC and Keegan, KP and Allada, R}, Authors = {Pfeiffenberger, C and Lear, BC and Keegan, KP and Allada, R},

Volume = {2010}, Year = {2010}} @article{11291099, Author = {Kitamoto, T}, Authors = {Kitamoto, T}, Date = {2001 May}, Journal = {J Neurobiol}, Month = {May}, Pages = {81-92}, Pubmed_Id = {11291099}, Source = {J Neurobiol}, Title = {{Conditional modification of behavior in Drosophila by targeted expression of a temperature-sensitive shibire allele in defined neurons.}}, Volume = {47}, Year = {2001}} @article{12568241, Author = {Hendricks, JC and Lu, S and Kume, K and Yin, JC and Yang, Z and Sehgal, A}, Authors = {Hendricks, JC and Lu, S and Kume, K and Yin, JC and Yang, Z and Sehgal, A},

Volume = {60}, Year = {2008}} @article{23864373, Author = {Iniguez, J and Schutte, SS and O'Dowd, DK}, Authors = {Iniguez, J and Schutte, SS and O'Dowd, DK}, Date = {2013 Oct}, Journal = {J Neurophysiol}, Month = {Oct}, Pages = {1490-6}, Pubmed_Id = {23864373}, Source = {J Neurophysiol}, Title = {{Cav3-type α1T calcium channels mediate transient calcium currents that regulate repetitive firing in Drosophila antennal lobe PNs.}}, Volume = {110}, Year = {2013}} @article{23849427, Author = {Park, HJ and Park, SJ and Ahn, EJ and Lee, SY and Seo, H and Lee, JH}, Authors = {Park, HJ and Park, SJ and Ahn, EJ and Lee, SY and Seo, H and Lee, JH},

Title = {{Asp residues of the Glu-Glu-Asp-Asp pore filter contribute to ion permeation and selectivity of the Ca(v)3.2 T-type channel.}}, Volume = {54}, Year = {2013}} @article{20187142, Author = {Young, JM and Armstrong, JD}, Authors = {Young, JM and Armstrong, JD}, Date = {2010 May 1}, Journal = {J Comp Neurol}, Month = {May}, Pages = {1500-24}, Pubmed_Id = {20187142}, Source = {J Comp Neurol}, Title = {{Structure of the adult central complex in Drosophila: organization of distinct neuronal subsets.}}, Volume = {518}, Year = {2010}}

In this study, we cloned voltage gated T-type Ca\textsuperscript{2+} channel in Drosophila (DmCa\textsubscript{v}3). The protein size of DmCa\textsubscript{v}3 is 3205 amino acids, the largest T-type channel cloned to date\cite{senatore:2010aa}. Electrophysiological characterization of DmCa\textsubscript{v}3 showed that the channel has the hallmark properties of native T-type channels described from isolated cells as well as those of cloned T-type channel; 1) low-threshold activation at around \textminus60 mV and creation of maximal current amplitude at \textminus20 mV in the I-V plot, 2) transient current kinetics during step pulses producing a criss-crossing pattern between current traces elicited by an I-V protocol and 3) slow deactivation of tail currents (Fig. \ref{fig:1}). These results indicate that DmCa\textsubscript{v}3 is a T-type channel cloned from \emph{Drosophila melanogaster}. Biophysical properties of the cloned channel are consistent with previous studies showing that DmCa\textsubscript{v}3 mediates low-voltage-activated (LVA) currents both in central and peripheral nervous system\cite{Ryglewski:2012jk, 23864373}. Iniguez:2013ib}. Analysis of the fly genome suggests that DmCa\textsubscript{v}3 is unique T-type channel isoform in the fly, which is different from existence of the three isoforms (Ca\textsubscript{v}3.1-3.3) in mammals. In current kinetics of view, DmCa\textsubscript{v}3 is similar to mammalian Ca\textsubscript{v}3.1 and Ca\textsubscript{v}3.2 rather than mammalian Ca\textsubscript{v}3.3 with much slower current kinetics. In terms of relative permeability and nickel inhibition sensitivity, the preferential permeation of Ba\textsuperscript{2+} over Ca\textsuperscript{2+} and nickel sensitive inhibition of DmCa\textsubscript{v}3 suggest that the fly T-type channel resembles to mammalian Ca\textsubscript{v}3.2\cite{16377633, 23849427}.

Flies were kept on a standard corn meal, corn syrup, yeast and agar medium at room temperature. \href{http://flystocks.bio.indiana.edu/Reports/27392.html}{\emph{UAS-mCD8-ChRFP}}, \href{http://flystocks.bio.indiana.edu/Reports/26160.html}{\emph{Vglut-gal4}} and \emph{Gad1-gal4} (47140) was obtained from \href{http://flystocks.bio.indiana.edu}{Bloomington Drosophila Stock Center} (Indiana, USA). \emph{c465-gal4} and \emph{210y-gal4} were gifts from J. Douglas Armstrong\cite{20187142}. Armstrong\cite{Young:2010jq}. The following stocks were described previously: \emph{Elav-gal4}\cite{Lin:1994vn}, \emph{Cha-gal4}\cite{11291099} \emph{Cha-gal4}\cite{Kitamoto:2001ue} \emph{c161-gal4}\cite{renn:1999aa}, \emph{104y-gal4}\cite{sakai:2006aa}, \emph{c309-gal4}\cite{connolly:1996aa}, \emph{MB247-gal4}\cite{zars:2000aa}, \emph{pdf-gal4}\cite{renn:1999ab}, \emph{TH-gal4}\cite{friggi-grelin:2003aa}, \emph{Tdc2-gal4}\cite{alekseyenko:2010aa}, \emph{c232-gal4}\cite{renn:1999aa}, \emph{TRH-gal4}\cite{alekseyenko:2010aa}, \emph{GMR-gal4}\cite{freeman:1996aa}, \emph{c929-gal4}\cite{taghert:2001aa}, \emph{clk8.0-gal4}\cite{glossop:2003aa} and \emph{Dilp2-gal4}\cite{Rulifson:2002cg}.