Nature Protocols, Год журнала: 2017, Номер 12(11), С. 2355 - 2359
Опубликована: Окт. 12, 2017
Язык: Английский
G3 Genes Genomes Genetics, Год журнала: 2018, Номер 9(1), С. 81 - 94
Опубликована: Ноя. 6, 2018
The insect mushroom body (MB) is a conserved brain structure that plays key roles in diverse array of behaviors. Drosophila melanogaster MB the primary invertebrate model neural circuits related to memory formation and storage, its development, morphology, wiring, function has been extensively studied. MBs consist intrinsic Kenyon Cells are divided into three major neuron classes (γ, α'/β' α/β) 7 cell subtypes (γd, γm, α'/β'ap, α'/β'm, α/βp, α/βs α/βc) based on their birth order, connectivity. These play distinct processing, however underlying transcriptional differences unknown. Here, we used RNA sequencing (RNA-seq) profile nuclear transcriptomes each neuronal subtypes. We identified 350 class- or subtype-specific genes, including widely α/β class marker Fas2 trio Immunostaining corroborates RNA-seq measurements at protein level for several cases. Importantly, our data provide full accounting neurotransmitter receptors, transporters, biosynthetic enzymes, neuropeptides, neuropeptide receptors expressed within these types. This high-quality, type-level transcriptome catalog provides valuable resource fly neuroscience community.
Язык: Английский
Процитировано
53
eLife, Год журнала: 2018, Номер 7
Опубликована: Фев. 9, 2018
Sleep during development is involved in refining brain circuitry, but a role for sleep the earliest periods of nervous system elaboration, when neurons are first being born, has not been explored. Here we identify state Drosophila larvae that coincides with major wave neurogenesis. Mechanisms controlling larval partially distinct from adult sleep: octopamine, analog mammalian norepinephrine, arousal neuromodulator larvae, dopamine required. Using real-time behavioral monitoring closed-loop deprivation system, find loss impairs cell division neural progenitors. This work establishes uniquely suited studying nascent periods, and demonstrates early life regulates stem proliferation.
Язык: Английский
Процитировано
49
European Journal of Neuroscience, Год журнала: 2018, Номер 51(1), С. 454 - 481
Опубликована: Сен. 30, 2018
Abstract Endogenous circadian oscillators regulate molecular, cellular and physiological rhythms, synchronizing tissues organ function to coordinate activity metabolism with environmental cycles. The technological nature of modern society round‐the‐clock work schedules heavy reliance on personal electronics has precipitated a striking increase in the incidence sleep disorders. Circadian dysfunction contributes an increased risk for many diseases appears have adverse effects aging longevity animal models. From invertebrate organisms humans, synchronization system weakens age aggravating age‐related disorders pathologies. In this review, we highlight impacts reciprocal examples from Drosophila humans underscoring highly conserved these interactions. Additionally, review potential using reinforcement promote healthy mitigate Advancements medicine public health significantly human life span past century. With demographics countries worldwide shifting older population, there is critical need understand factors that shape aging. melanogaster , as model interactions, capacity facilitate rapid advancement research area provide mechanistic insights targeted investigations mammals.
Язык: Английский
Процитировано
48
EMBO Reports, Год журнала: 2019, Номер 20(3)
Опубликована: Фев. 25, 2019
Review25 February 2019Open Access Genetic sleep deprivation: using mutants to study functions Henrik Bringmann Corresponding Author [email protected] orcid.org/0000-0002-7689-8617 Max Planck Institute for Biophysical Chemistry, Göttingen, Germany Search more papers by this author Information *,1 1Max *Corresponding author. Tel: +49 551 2011091; E-mail: EMBO Reports (2019)20:e46807https://doi.org/10.15252/embr.201846807 See the Glossary abbreviations used in article. PDFDownload PDF of article text and main figures. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Sleep is a fundamental conserved physiological state animals humans. It may serve multiple functions, ranging from energy conservation higher brain operation. Understanding underlying mechanisms requires sleeplessness its consequences. The traditional approach remove deprivation (SD) sensory stimulation. However, stimulation-induced SD can be stressful cause non-specific side effects. An emerging alternative method "genetic SD", which removes genetics or optogenetics. sleep-active neurons their regulators. Thus, genetic impairment circuits might lead specific comprehensive loss. Here, I discuss advantages limits key model animals: rodents, zebrafish, fruit flies roundworms, how alters our view functions. typically causes less severe phenotypes compared with SD, suggesting that have overestimated role sleep, calling re-investigation ALA name Caenorhabditis elegans interneuron/mechanosensory neuron AMPA α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid CNO clozapine-N-oxide dFB dorsal fan-shaped body EEG electroencephalogram EGF epidermal growth factor GABA γ-aminobutyric GPCR G protein-coupled receptor HPA hypothalamic–pituitary–adrenal axis MS medial septum PB parabrachial nuclei PI pars intercerebralis PZ parafacial zone REM rapid eye movement RIS ring interneuron s, C. ROS reactive oxygen species SIK3 salt-inducible kinase 3 VLPO ventrolateral preoptic nucleus Introduction We spend about one-third lives asleep. curtailment detrimental human health associated an increased risk infection, depression, cardiovascular disease, obesity, type 2 diabetes, cancer, illustrating importance time investment. high prevalence insomnia insufficient quality modern societies thus presents massive unmet problem 1234. appears affect virtually all aspects animal physiology, numerous processes been proposed depend on regular occurrence sleep. While broad consequences loss are obvious, there still no consensus as what direct are, carries out at molecular level. research first focused Seminal work recordings showed humans two types non-REM also called active quiet respectively. During shows asynchronous activity similar wake, concomitant paralysis striated muscles, few exceptions including musculature controlling breathing. both muscles show reduced highly synchronous 56. Using electrophysiological characteristics it has possible detect wide range mammals birds 78. Even reptiles possess different states resemble whereas amphibians appear only 9. This led conclusion diverged base amniotes into Behavioral quiescence long observed across species, invertebrates. defining correlates often not due anatomical differences. Nevertheless, identified four behavioral criteria 10. (i) A typical posture assumed compatible muscle activity. (ii) reduces responsiveness stimulation, indicating global neural dampening extends systems, contrasts wakefulness. (iii) rapidly reversible, meaning readily awoken, separating coma paralysis. (iv) under homeostatic regulation, implying exist ensure takes place, underscoring By applying these criteria, nervous system, cnidaria presenting most basal phylum detected 11. much widespread among than initially thought 12. Particularly important was identification three non-mammalian models, Drosophila, elegans, models facilitate mechanistic dissection 1314151617. Compelling evidence existence system but never sleeps lacking, amount plastic some get away little Environmental conditions impact need, spent differs substantially species. Under extreme conditions, temporary restriction even complete confers selective advantage. For example, migrating mating able suspend reduce need least several days 1819. Also, such large herbivores cave-dwelling fish, manage live sleeping little, h per day sufficient 2021. On other extreme, bats up 20 21. suggests adapted to, depends ecological constraints, perhaps regulate behavior preserve 22. Because asleep 10% time, lower limit required survival seems (Fig 1). Figure 1. fraction varies greatly does drop below 10%Sleep between 3–20 h/24 lot Model organisms fall within wild 3885103124. Download figure PowerPoint Functions underpinnings play roles coarsely sorted groups overlapping mutually exclusive. group function theories posits plays optimizing allocation energy. second core cellular processes. And third serves 1223 2). 2. Summary hypothesized sleepVarious ideas changes hypotheses depicted here. (A) In simple form, save when adaptive. would hibernation Energy saved later use could rather allocated anabolic reactions protein synthesis 25. (B) become adaptive compartmentalizing conflicting metabolic make efficient 36. controls hormones, food intake, metabolism (including lipid sugar metabolism) 34. cyclic biochemical reactions. Wakefulness, phosphorylation synaptic proteins dephosphorylation 37. Various homeostasis exist, accumulation extracellular adenosine 144. immune 323334. (C) plasticity learning memory. Synaptic during include downscaling weak synapses, process promoted Homer1a. Strong synapses preserved 4547145. support systems memory consolidation re-activating re-distributing areas 49. These re-arrangements novel insight creativity 50. Note overlapping. Most stems value understood viewing inactive state. At times wakefulness advantageous, organism enter strong argument energetic constraints determining variation intensity seen share energy-saving torpor, metabolically behaviorally phase found characterized temperature, instance hibernation. Both transitions torpor well exit involve they related 222425. differ defined reversible state, generally reversible. functional difference dissipate 2627. benefits cannot simply explained alone. According theory primarily diverted restorative biosynthetic 2528. becomes regenerative allowing facilitating housekeeping supported transcriptomic studies increase expression genes biosynthesis transport 293031. Anabolic could, growth, stress resistance, 32333435. control metabolism, part, regulating rhythmic timing intake. instance, increases concentration appetite-stimulating hormone ghrelin, appetite-inhibiting leptin, syndrome, diabetes itself present cycle, provides temporal compartmentalization difficult reconcile energetically favorable if carried subsequently example cycling reaction substantial proteins, globally wakefulness, decreased responsible cycle SIK3, gain-of-function mutation sleepy excessive duration 38. known regulator link 3940. Completing picture housekeeping, mice period potentially toxic molecules aggregates removed brain. removal neuronal shrinkage increasing flux interstitial fluid 41. experiments good night's Memory formation regions encode Transcriptome analysis brains formation, 424344. Plasticity involves alterations size composition synapses. new memories strengthen form weaken disappear. leads net synapse subsequent downscaling, mostly affects leaves intact 45. weakening receptors, Homer1a 46. working model, kept noradrenergic signaling enters recruitment adenosine, somnogenic (sleep-promoting) accumulate promotes drive 4748. Besides cell biological composition, occurs level involving recurrent reactivation redistribution integration existing circuits, updating knowledge. Disconnecting input restructuring mature allow associations creative insights problems hard solve help consolidate certain memories, that, mediated hippocampus, though 649. induced crucial induction. Research substrates started patients who had suffered consequence infection-induced injury. Lesions particular area, anterior hypothalamus, reduction demonstrating dedicated centers mammalian 51. motivated centers, cats, rats, mice. Central induction sleep-promoting express inhibitory neurotransmitters, GABA, neuropeptides. Sleep-active depolarize specifically onset inhibit wake-promoting promote inhibited stimulation arousal quick reversibility. They over-activated confer drive. motor turn regulated upstream driver determine 5253. reveals regarding based observations correlate causality established studying deprivation. wakefulness-promoting oppose each generate discrete 54. i.e., leading inhibition system. Stimulation-induced accounts causal testing summarized above. Acute essential Complete rodents caused weight loss, skin ulceration, sepsis, ultimately death experimental 55. To prevent lethality, applied partially shorten then restriction, imposed chronically Chronic understand effects chronic health. injury vigilance attribute partial stress. pleiotropic made impossible clearly deduce immediate Sleep, arousal, intimately linked hyperarousal mental mammals, activates sets off response, maintains suppresses perpetuating vicious 5758. Gentler protocols standard today aim arouse motivating instead stressing. achieved over pathways 3) 59. confounding after interference rebound wake pressure "lapses" "microsleep" bouts disturb deficits attention, memory, information processing 60. performance, whether defects directly mechanisms. 3. Classic impairs sleep-inducing systemAccording flip-flop switch antagonizing wake-inducing 52. dominates wake. Sensory suppressing despite (D) Genetically impairing permits disinhibition. contribute part normal waking ablation hyperarousal. arousing effect likely smaller hyperactivity manipulations without causing hyperactivity. approaches change fundamentally means complementary. should pursued establishing strategy render sleepless paradigm, lacks induction, requiring additional following attributable disinhibition 3). How impaired? ablate parts neurosurgical methods, way impair through targeting. call SD". deletion results lethality many either conditional approaches. Conditional generated optogenetic chemogenetic inducible knockouts create analog Alternatively, hypomorphic mutations analogs restriction. imminently lethal, choice phenotypes. As targeting directly, manipulating downstream employed removing achieved, activating preventing normally neurons. complement studies, devised activate gain". Specificity mutant phenotype indirectly. circadian rhythms abrogation result 6162. phenotype, will Similarly, strongly same 63. probably obtained mutating circuits. arousal. Assuming minor limiting prominent inducing moderate Consistent idea, (see below). specificity systems. degree long-term consequences, genetically tools organisms, high, complete. Homeostatic compensatory compensate depth remaining which, ameliorates Some relatively small amounts fulfill sleep's 2152. detectable residual advantageous bound. induces over-activation neurons, baseline 5464. systems: discoveries were variety mice, monkeys. pivotal structures turned conserved. Its amenability mouse intensively 236566. investigated mammals. rodent Forward screening Dreamless, dominant gene encodes ion channel excitability, GABAergic ventral medulla stem. Inhibition induce optogenetically depolarizing 67. Dreamless investigate yet published. Proving challenge because 6. involved high-amplitude theta waves hippocampal EEG. itself, silenced Silencing providing 68. optogenetics Mutants completely described, display very ideal 6269. gain 4). There principal triggering via successfully rodents. impaired. area PB, activated chemogenetically extend restrict 70. Alternatively extended supramammillary glutamatergic 71. lesioning approximately 50% stress, hyperarousal, 7273. controlled 74. Sleep-promoting indirectly, activation inhibitors lateral hypothalamus forebrain 7576. Other forebrain, stem, cortex contain 66. brainstem region shown sleep-active, 40%, 5) 777879. populations explain why subsets fascinating ablating reasonable next step combine ablations see stronger 4. modelsShown examples low specificity, levels do
Язык: Английский
Процитировано
45
Nature Protocols, Год журнала: 2017, Номер 12(11), С. 2355 - 2359
Опубликована: Окт. 12, 2017
Язык: Английский
Процитировано
48