Optical calcium imaging in the nervous system of Drosophila melanogaster

Riemensperger, Thomas; Pech, Ulrike; Dipt, Shubham; Fiala, André

doi:10.1016/j.bbagen.2012.02.013

Cited by 32 publications

(25 citation statements)

References 127 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Other aspects of functional connectomics, such as imaging (e.g. Riemensperger et al ., 2012) and manipulating (Simpson, 2009) neural activity, have been well reviewed elsewhere and will only be updated here. Given that this is a relatively new field, we will include not only the methods used to trace synaptic circuits in the fly but also those originally described in other systems with potential Drosophila applications.…”

Section: Introductionmentioning

confidence: 99%

The Genetic Analysis of Functional Connectomics in Drosophila

Meinertzhagen

Lee

2012

Advances in Genetics

View full text Add to dashboard Cite

Fly and vertebrate nervous systems share many organization characteristics, such as layers, columns and glomeruli, and utilize similar synaptic components, such ion channels and receptors. Both also exhibit similar network features. Recent technological advances, especially in electron microscopy, now allow us to determine synaptic circuits and identify pathways cell-by-cell, as part of the fly’s connectome. Genetic tools provide the means to identify synaptic components, as well as to record and manipulate neuronal activity, adding function to the connectome. This review discusses technical advances in these emerging areas of functional connectomics, offering prognoses in each and identifying the challenges in bridging structural connectomics to molecular biology and synaptic physiology, thereby determining fundamental computation mechanisms that underlie behaviour.

show abstract

Section: Introductionmentioning

confidence: 99%

The Genetic Analysis of Functional Connectomics in Drosophila

Meinertzhagen

Lee

2012

Advances in Genetics

View full text Add to dashboard Cite

show abstract

“…Thus, we also asked whether this phenomenon occurs in flies. We focused on the MBs, large areas of the Drosophila brain involved in olfactory learning (16, 17) and sleep regulation (18,19), and used in vivo calcium imaging (20,21) to measure spontaneous and evoked neuronal activity in the MB principal neurons, the Kenyon cells, during sleep, wake, and in response to different periods of sleep deprivation. …”

mentioning

confidence: 99%

Sleep- and wake-dependent changes in neuronal activity and reactivity demonstrated in fly neurons using in vivo calcium imaging

Bushey

Tononi

Cirelli

2015

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Sleep in Drosophila shares many features with mammalian sleep, but it remains unknown whether spontaneous and evoked activity of individual neurons change with the sleep/wake cycle in flies as they do in mammals. Here we used calcium imaging to assess how the Kenyon cells in the fly mushroom bodies change their activity and reactivity to stimuli during sleep, wake, and after short or long sleep deprivation. As before, sleep was defined as a period of immobility of >5 min associated with a reduced behavioral response to a stimulus. We found that calcium levels in Kenyon cells decline when flies fall asleep and increase when they wake up. Moreover, calcium transients in response to two different stimuli are larger in awake flies than in sleeping flies. The activity of Kenyon cells is also affected by sleep/wake history: in awake flies, more cells are spontaneously active and responding to stimuli if the last several hours (5-8 h) before imaging were spent awake rather than asleep. By contrast, long wake (≥29 h) reduces both baseline and evoked neural activity and decreases the ability of neurons to respond consistently to the same repeated stimulus. The latter finding may underlie some of the negative effects of sleep deprivation on cognitive performance and is consistent with the occurrence of local sleep during wake as described in behaving rats. Thus, calcium imaging uncovers new similarities between fly and mammalian sleep: fly neurons are more active and reactive in wake than in sleep, and their activity tracks sleep/wake history.T he fundamental features that characterize mammalian sleep also define Drosophila melanogaster sleep (1-3). Most crucially, in both flies and mammals, sleep is distinguished from simple rest (quiet wake) by an increased arousal threshold, i.e., a reduced ability to respond to external stimuli. Moreover, in flies and mammals sleep is controlled homeostatically by the duration as well by the intensity of prior wake, suggesting basic similarities in the mechanisms of sleep regulation across species. Thus, in flies both sleep deprivation and a rich learning experience lead to a sleep rebound characterized by overall increased sleep time, increased arousal threshold, and longer sleep episodes (4-6). As in mammals, overall neuronal activity in flies is also high during wake and low during sleep (6-8). Specifically, a seminal study using local field potential (LFP) recordings from the Drosophila medial protocerebrum found high spike-like potentials that disappeared after the block of synaptic transmission in the mushroom bodies (MBs) (7). This high spike activity was present when flies were moving or had been quiescent for only a few seconds, but disappeared with sleep (i.e., after periods of immobility >5 min), when the overall LFP power in all frequencies also decreased by ∼60% (7). The study concluded that neural activity in the sleeping fly brain, or at least in a central region spanning the MBs, resembles that seen in mammals in several brainstem cell groups including noradrenergic ne...

show abstract

“…The intracellular Ca 2+ level closely correlates with neuronal excitation (Berridge, 1998; Burgoyne, 2007). Optical Ca 2+ imaging represents, therefore, a widely used technique to monitor the activity of neurons in general (Grienberger and Konnerth, 2012), and also in the central brain of Drosophila (Riemensperger et al, 2012). We first created transgenic flies expressing the widely-used fluorescence Ca 2+ sensors GCaMP3.0 (Tian et al, 2009) under control of the mb247 promoter and performed two-photon optical Ca 2+ imaging experiments with the focus on the horizontal lobes.…”

Section: Resultsmentioning

confidence: 99%

“…Transgenes that can help to analyze neuronal structure and/or function are, first, anatomical markers, e.g., cytosolic or subcellular anchored fluorescence proteins. Second, reporter proteins can be expressed to monitor physiological parameters of neuronal function, e.g., intracellular Ca 2+ dynamics (Fiala et al, 2002; Riemensperger et al, 2012) or second-messenger signaling (Lissandron et al, 2007; Shafer et al, 2008). Third, effector proteins can be expressed to manipulate specific aspects of neuronal functioning.…”

Section: Introductionmentioning

confidence: 99%

Mushroom body miscellanea: transgenic Drosophila strains expressing anatomical and physiological sensor proteins in Kenyon cells

Pech¹,

Dipt²,

Barth³

et al. 2013

Front. Neural Circuits

Self Cite

View full text Add to dashboard Cite

The fruit fly Drosophila melanogaster represents a key model organism for analyzing how neuronal circuits regulate behavior. The mushroom body in the central brain is a particularly prominent brain region that has been intensely studied in several insect species and been implicated in a variety of behaviors, e.g., associative learning, locomotor activity, and sleep. Drosophila melanogaster offers the advantage that transgenes can be easily expressed in neuronal subpopulations, e.g., in intrinsic mushroom body neurons (Kenyon cells). A number of transgenes has been described and engineered to visualize the anatomy of neurons, to monitor physiological parameters of neuronal activity, and to manipulate neuronal function artificially. To target the expression of these transgenes selectively to specific neurons several sophisticated bi- or even multipartite transcription systems have been invented. However, the number of transgenes that can be combined in the genome of an individual fly is limited in practice. To facilitate the analysis of the mushroom body we provide a compilation of transgenic fruit flies that express transgenes under direct control of the Kenyon-cell specific promoter, mb247. The transgenes expressed are fluorescence reporters to analyze neuroanatomical aspects of the mushroom body, proteins to restrict ectopic gene expression to mushroom bodies, or fluorescent sensors to monitor physiological parameters of neuronal activity of Kenyon cells. Some of the transgenic animals compiled here have been published already, whereas others are novel and characterized here for the first time. Overall, the collection of transgenic flies expressing sensor and reporter genes in Kenyon cells facilitates combinations with binary transcription systems and might, ultimately, advance the physiological analysis of mushroom body function.

show abstract

Optical calcium imaging in the nervous system of Drosophila melanogaster

Cited by 32 publications

References 127 publications

The Genetic Analysis of Functional Connectomics in Drosophila

The Genetic Analysis of Functional Connectomics in Drosophila

Sleep- and wake-dependent changes in neuronal activity and reactivity demonstrated in fly neurons using in vivo calcium imaging

Mushroom body miscellanea: transgenic Drosophila strains expressing anatomical and physiological sensor proteins in Kenyon cells

Contact Info

Product

Resources

About