Temporal Calcium Profiling of Specific Circadian Neurons in Freely Moving Flies

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2017 Oct 5

PMID 28973886

Citations 44

Authors

Fang Guo

Xiao Chen

Michael Rosbash

Affiliations

Soon will be listed here.

Abstract

There are no general methods for reliably assessing the firing properties or even calcium profiles of specific neurons in freely moving flies. To this end, we adapted a GFP-based calcium reporter to luciferase that was expressed in small subsets of circadian neurons. This Tric-LUC reporter allowed a direct comparison of luciferase activity with locomotor activity, which was assayed in the same flies with video recording. The LUC profile from activity-promoting E cells paralleled evening locomotor activity, and the LUC profile from sleep-promoting glutamatergic DN1s (gDN1s) paralleled daytime sleep. Similar profiles were generated by novel reporters recently identified based on transcription factor activation. As E cell and gDN1 activity is necessary and sufficient for normal evening locomotor activity and daytime sleep profiles, respectively, we suggest that their luciferase profiles reflect their neuronal calcium and in some cases firing profiles in wake-behaving flies.

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References

Tataroglu O, Emery P . Studying circadian rhythms in Drosophila melanogaster. Methods. 2014; 68(1):140-50. PMC: 4049855. DOI: 10.1016/j.ymeth.2014.01.001. View

Liang X, Holy T, Taghert P . Synchronous Drosophila circadian pacemakers display nonsynchronous Ca²⁺ rhythms in vivo. Science. 2016; 351(6276):976-81. PMC: 4836443. DOI: 10.1126/science.aad3997. View

Tanenhaus A, Zhang J, Yin J . In vivo circadian oscillation of dCREB2 and NF-κB activity in the Drosophila nervous system. PLoS One. 2012; 7(10):e45130. PMC: 3471920. DOI: 10.1371/journal.pone.0045130. View

Flourakis M, Kula-Eversole E, Hutchison A, Han T, Aranda K, Moose D . A Conserved Bicycle Model for Circadian Clock Control of Membrane Excitability. Cell. 2015; 162(4):836-48. PMC: 4537776. DOI: 10.1016/j.cell.2015.07.036. View

Renn S, Park J, Rosbash M, Hall J, Taghert P . A pdf neuropeptide gene mutation and ablation of PDF neurons each cause severe abnormalities of behavioral circadian rhythms in Drosophila. Cell. 2000; 99(7):791-802. DOI: 10.1016/s0092-8674(00)81676-1. View

Yao Z, Shafer O . The Drosophila circadian clock is a variably coupled network of multiple peptidergic units. Science. 2014; 343(6178):1516-20. PMC: 4259399. DOI: 10.1126/science.1251285. View

Helfrich-Forster C . Differential control of morning and evening components in the activity rhythm of Drosophila melanogaster--sex-specific differences suggest a different quality of activity. J Biol Rhythms. 2000; 15(2):135-54. DOI: 10.1177/074873040001500208. View

Roberts L, Leise T, Noguchi T, Galschiodt A, Houl J, Welsh D . Light evokes rapid circadian network oscillator desynchrony followed by gradual phase retuning of synchrony. Curr Biol. 2015; 25(7):858-67. PMC: 4399721. DOI: 10.1016/j.cub.2015.01.056. View

Mohammad F, Stewart J, Ott S, Chlebikova K, Chua J, Koh T . Optogenetic inhibition of behavior with anion channelrhodopsins. Nat Methods. 2017; 14(3):271-274. DOI: 10.1038/nmeth.4148. View

10.

Zhang L, Chung B, Lear B, Kilman V, Liu Y, Mahesh G . DN1(p) circadian neurons coordinate acute light and PDF inputs to produce robust daily behavior in Drosophila. Curr Biol. 2010; 20(7):591-9. PMC: 2864127. DOI: 10.1016/j.cub.2010.02.056. View

11.

Parisky K, Agosto Rivera J, Donelson N, Kotecha S, Griffith L . Reorganization of Sleep by Temperature in Drosophila Requires Light, the Homeostat, and the Circadian Clock. Curr Biol. 2016; 26(7):882-92. PMC: 5080662. DOI: 10.1016/j.cub.2016.02.011. View

12.

Masuyama K, Zhang Y, Rao Y, Wang J . Mapping neural circuits with activity-dependent nuclear import of a transcription factor. J Neurogenet. 2012; 26(1):89-102. PMC: 3357894. DOI: 10.3109/01677063.2011.642910. View

13.

Kunst M, Hughes M, Raccuglia D, Felix M, Li M, Barnett G . Calcitonin gene-related peptide neurons mediate sleep-specific circadian output in Drosophila. Curr Biol. 2014; 24(22):2652-64. PMC: 4255360. DOI: 10.1016/j.cub.2014.09.077. View

14.

Levine J, Funes P, Dowse H, Hall J . Signal analysis of behavioral and molecular cycles. BMC Neurosci. 2002; 3:1. PMC: 65508. DOI: 10.1186/1471-2202-3-1. View

15.

Zhang Y, Liu Y, Bilodeau-Wentworth D, Hardin P, Emery P . Light and temperature control the contribution of specific DN1 neurons to Drosophila circadian behavior. Curr Biol. 2010; 20(7):600-5. PMC: 2862552. DOI: 10.1016/j.cub.2010.02.044. View

16.

Seluzicki A, Flourakis M, Kula-Eversole E, Zhang L, Kilman V, Allada R . Dual PDF signaling pathways reset clocks via TIMELESS and acutely excite target neurons to control circadian behavior. PLoS Biol. 2014; 12(3):e1001810. PMC: 3958333. DOI: 10.1371/journal.pbio.1001810. View

17.

Inagaki H, Jung Y, Hoopfer E, Wong A, Mishra N, Lin J . Optogenetic control of Drosophila using a red-shifted channelrhodopsin reveals experience-dependent influences on courtship. Nat Methods. 2013; 11(3):325-32. PMC: 4151318. DOI: 10.1038/nmeth.2765. View

18.

Cavanaugh D, Geratowski J, Wooltorton J, Spaethling J, Hector C, Zheng X . Identification of a circadian output circuit for rest:activity rhythms in Drosophila. Cell. 2014; 157(3):689-701. PMC: 4003459. DOI: 10.1016/j.cell.2014.02.024. View

19.

Yoshii T, Vanin S, Costa R, Helfrich-Forster C . Synergic entrainment of Drosophila's circadian clock by light and temperature. J Biol Rhythms. 2009; 24(6):452-64. DOI: 10.1177/0748730409348551. View

20.

Guo F, Cerullo I, Chen X, Rosbash M . PDF neuron firing phase-shifts key circadian activity neurons in Drosophila. Elife. 2014; 3. PMC: 4092873. DOI: 10.7554/eLife.02780. View