Medicine in the Fourth Dimension

Cell Metabolism, Journal Year: 2019, Volume and Issue: 30(2), P. 238 - 250

Published: Aug. 1, 2019

Language: Английский

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The cognitive impact of light: illuminating ipRGC circuit mechanisms

Nature reviews. Neuroscience, Journal Year: 2024, Volume and Issue: 25(3), P. 159 - 175

Published: Jan. 26, 2024

Language: Английский

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The time is now: accounting for time-of-day effects to improve reproducibility and translation of metabolism research

Nature Metabolism, Journal Year: 2025, Volume and Issue: unknown

Published: March 17, 2025

Language: Английский

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A Head-Mounted Camera System Integrates Detailed Behavioral Monitoring with Multichannel Electrophysiology in Freely Moving Mice

Neuron, Journal Year: 2018, Volume and Issue: 100(1), P. 46 - 60.e7

Published: Oct. 1, 2018

Highlights•Eyes, whiskers, head, and neural activity monitored in freely moving mice•System generates stable video output leaves mouse behavior largely unchanged•Close link between eye head movements at both slow fast timescales•Active the dark strongly modulate primary visual cortex activitySummaryBreakthroughs understanding basis of natural require recording intervention to be paired with high-fidelity multimodal behavioral monitoring. An extensive genetic toolkit for circuit dissection, well-developed technology, make a powerful model organism systems neuroscience. However, most methods high-bandwidth acquisition data mice rely upon fixed-position cameras other off-animal devices, complicating monitoring animals engaged behaviors. Here, we report development lightweight head-mounted camera system combined head-movement sensors simultaneously monitor position, pupil dilation, whisking, pinna along motion unrestrained, behaving mice. The power technology is demonstrated by observations linking position orientation; whisking non-tactile stimulation; and, electrophysiological experiments, cortical volitional movements.

Language: Английский

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Regulation and function of neurogenesis in the adult mammalian hypothalamus

Progress in Neurobiology, Journal Year: 2018, Volume and Issue: 170, P. 53 - 66

Published: April 6, 2018

Language: Английский

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Circadian Photoentrainment in Mice and Humans

Biology, Journal Year: 2020, Volume and Issue: 9(7), P. 180 - 180

Published: July 21, 2020

Light around twilight provides the primary entrainment signal for circadian rhythms. Here we review mechanisms and responses of mouse human systems to light. Both utilize a network photosensitive retinal ganglion cells (pRGCs) expressing photopigment melanopsin (OPN4). In both species action spectra functional expression OPN4 in vitro show that has λmax close 480 nm. Anatomical findings demonstrate there are multiple pRGC sub-types, with some evidence mice, but little humans, regarding their roles regulating physiology behavior. Studies non-human primates rods cones project can modulate light pRGCs. Such an integration signals enables detect dim light, higher intensities intermittent exposure, whilst measures bright over extended periods time. Although photoreceptor similar, sensitivity thresholds differ markedly between mice humans. Mice entrain at approximately 1 lux few minutes, humans require high irradiance (>100's lux) long duration (>30 min). The basis this difference remains unclear. As our exposure is highly dynamic, because interactions complex difficult model, attempts develop evidence-based lighting enhance very challenging. A way forward will be define artificial natural "real world" where intensity, duration, spectral quality, time day, history age each assessed.

Language: Английский

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Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning

Scientific Reports, Journal Year: 2018, Volume and Issue: 8(1)

Published: May 18, 2018

We present a miniature head mounted two-photon fiber-coupled microscope (2P-FCM) for neuronal imaging with active axial focusing enabled using electrowetting lens. show three-dimensional of structure and record activity from GCaMP6s fluorescence multiple focal planes in freely-moving mouse. Two-color simultaneous GFP tdTomato is also demonstrated. Additionally, dynamic control the scanning lens allows tilting plane enabling neurons depths to be imaged single plane. Two-photon increased penetration depth tissue yielding working distance 450 μm an additional 180 focusing. The objective NA 0.45 lateral resolution 1.8 μm, 10 field-of-view 240 diameter. 2P-FCM has weight only ~2.5 g capable repeatable stable head-attachment. provides new capability functionally distinct layers, opening opportunities neuroscience research.

Language: Английский

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Neurovascular coupling and bilateral connectivity during NREM and REM sleep

eLife, Journal Year: 2020, Volume and Issue: 9

Published: Oct. 29, 2020

To understand how arousal state impacts cerebral hemodynamics and neurovascular coupling, we monitored neural activity, behavior, hemodynamic signals in un-anesthetized, head-fixed mice. Mice frequently fell asleep during imaging, these sleep events were interspersed with periods of wake. During both NREM REM sleep, mice showed large increases blood volume ([HbT]) arteriole diameter relative to the awake state, two five times larger than those evoked by sensory stimulation. NREM, amplitude bilateral low-frequency oscillations [HbT] increased markedly, coherency between activity was higher resting states. Bilateral correlations highest lowest state. Hemodynamic cortex are strongly modulated changes substantially sensory-evoked responses.

Language: Английский

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Novel AAV capsids for intravitreal gene therapy of photoreceptor disorders

EMBO Molecular Medicine, Journal Year: 2021, Volume and Issue: 13(4)

Published: Feb. 22, 2021

Article22 February 2021Open Access Source DataTransparent process Novel AAV capsids for intravitreal gene therapy of photoreceptor disorders Marina Pavlou Department Ophthalmology, Ludwig-Maximilians-University, Munich, Germany Centre Integrated Protein Science Munich (CIPSM) at the Pharmacy, Search more papers by this author Christian Schön Laurence M Occelli Small Animal Clinical Sciences, Michigan State University, East Lansing, MI, USA Axel Rossi Laboratory Infection Biology and Gene Transfer, Institute Experimental Haematology, Hannover Medical School, Hannover, Nadja Meumann REBIRTH Research Translational Regenerative Medicine, Ryan F Boyd Ophthalmology Services, Charles River Laboratories, Mattawan, Joshua T Bartoe Jakob Siedlecki Maximilian J Gerhardt Sabrina Babutzka Jacqueline Bogedein Johanna E Wagner Siegfried G Priglinger Martin Biel Simon Petersen-Jones Hildegard Büning Corresponding Author [email protected] orcid.org/0000-0002-3365-6933 Stylianos Michalakis orcid.org/0000-0001-5092-9238 Information Pavlou1,2, Schön2, Occelli3, Rossi4, Meumann4,5, Boyd6, Bartoe6, Siedlecki1, Gerhardt1, Babutzka1,2, Bogedein1,2, Wagner2, Priglinger1, Biel2, Petersen-Jones3, *,4,5 *,1,2 1Department 2Centre 3Department 4Laboratory 5REBIRTH 6Ophthalmology *Corresponding author. Tel: +49 5115325106; E-mail: 89440053083; EMBO Mol Med (2021)13:e13392https://doi.org/10.15252/emmm.202013392 PDFDownload PDF article text main figures. Peer ReviewDownload a summary editorial decision including letters, reviewer comments responses to feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract using recombinant adeno-associated virus (rAAV) vectors treat blinding retinal dystrophies has become clinical reality. Therapeutically impactful targeting photoreceptors still relies on subretinal vector delivery, which detaches retina harbours substantial risks collateral damage, often without achieving widespread transduction. Herein, we report development novel engineered rAAV that enable efficient via less invasive administration. A unique in vivo selection procedure was performed, where an AAV2-based peptide-display library intravenously administered mice, followed isolation DNA from target cells after only 24 h. This stringent yielded vectors, termed AAV2.GL AAV2.NN, mediate high-level transduction injection dogs non-human primates. Importantly, both efficiently transduce human explant cultures. As proof-of-concept, Cnga3 delivery lead cone-specific expression protein rescued photopic cone Cnga3−/− mouse model achromatopsia. These expand applicability dystrophies. Synopsis Ocular aims improve or preserve vision patients with inherited disorders. The current technology administration therapeutic damage treats small portion affected retina. study presents two viral capable through route. AAV2.NN achieve mouse, dog primate single delivery. have rod tropism. outperforms existing mice. supplementation rescues function CNGA3 paper explained Problem Loss is one most severe handicaps high socioeconomic importance. More than 5 million suffer dystrophies, making them leading cause blindness young individuals. first specific form now clinically available. However, many other conditions caused mutations hundreds genes remain treatment. promising treatment concept supplement healthy copy mutated retina; requires use vector. Currently, standard practice delivering under patient's retina, effective but work addresses need better can light-sensing our efficiently, minimally route Results We optimised properties ocular therapy. show these efficacy three animal models simple intraocular also culture. For provide proof-of-concept data restoration total colour blindness. Impact could help patient outcome terms efficiency minimising evoked during make therapies successful accessible wider pool, as method simpler be performed any trained ophthalmologist. Introduction number obtaining market approval increasing, focus set improving first-generation tools safety. includes are considered tool acquired (IRDs) (Ali et al, 2017). Dependoparvovirus no assigned pathogenicity humans (Berns Muzyczka, lack integrase activity, proliferating well post-mitotic cells, long-term episomes slowly It therefore ideal pathologies manifesting neurons incapable innate regeneration. Many IRDs genetic degeneration photoreceptors. Consequently, supporting retinal-pigmented epithelium (RPE) developed restore inhibit delay pathological process. extensively used preclinical studies purpose, excellent safety record non-ocular (Trapani Auricchio, 2018). With regard eye, transient immune response been observed so far, did not limit success (Reichel 2018; Rabinowitz 2019). Indeed, supplementing RPE65 resulted authorization voretigene neparvovec RPE65-linked United States following year Europe (Keeler Flotte, In order outer far subretinal, entails surgical detachment RPE into temporally formed cavity, i.e. bleb. holds true neparvovec, uses serotype 2 (AAV2) one-off Despite its photoreceptors, deleterious already compromised exception certain (Khabou Boye 2020), conventional serotypes unable spread laterally local transduction, outside bleb area exposed sufficient amounts such, there unmet larger areas applicable route, such injection. recent years, various capsid engineering strategies employed (Grimm Büning, 2017) develop enhanced (Petrs-Silva 2009; Dalkara 2013; Zinn 2015; Katada study, combined systemic recovery genomes very pressure. Three screening rounds were vivo, C57BL6/J mice recovered cells. To further increase pressure, limited incubation time short period challenging detectable nuclei variants, across species produced yield, sensitive neutralising serum, injections primates (NHP), vitreal explants. Using achromatopsia (Biel 1999), validation restoring Library Our goal variants profile crossing biological barriers penetrance synaptic layers extracellular matrix. random AAV2 scaffold (Fig EV1A), 7-mer amino acid sequence inserted between N587 R588 EV1B). resulting approximately (Perabo 2003, 2006b) counter-selected wildtype heparan sulphate proteoglycan (HSPG) binding heparin affinity chromatography then injected adult (2-months old) > 8E10 (vg) initial (library #1) per mouse. At 24h injection, animals sacrificed, their retinas harvested, isolated. examined qPCR presence revealing accumulation thus barriers. Subsequently, amplified served template sublibrary production #2). repeated second round intravenous 5E11 vg time, isolated analysed reached internalised, successfully transported nucleus. #3) generated before, third round, #3 RG-eGFP old). magnetic-activated cell sorting (MACS) fluorescence-activated (FACS). had rods and/or cones extracted, NGS identify targeted cones. addition, whole samples #2. Click here figure. Figure EV1. Schematic diversification strategy Depiction parental structure (PDB 6IH9) VP3 monomer (amino acids (aa) 219–735) coloured blue. On right, blue shown unmodified insertion position I-587 circled residues highlighted orange yellow, respectively. constituent genes, assembly activating (AAP) (VP) 1–3. part VP3-coloured represents structural depicted (A) same colour. orange/yellow line schematic below indicate site peptides (orange) (yellow) flanked alanine linkers. Download figure PowerPoint Side-by-side comparison candidates Valid reads compared, valid means those containing encoding RGNAAA X1 X2 X3 X4 X5 X6 X7 AARQ "X" indicating Sequences detected excluded. Furthermore, ratio over calculated, select enriched favouring top five frequent AAV2.GL, AAV2.GA, AAV2.NS AAV2.SS (Table 1), vectorised tested displayed selected encoded self-complementary (sc) genome green fluorescent (eGFP) marker controlled ubiquitous cytomegalovirus (CMV) promoter. interrogate capacity 2E9 intravitreally 2-month-old (n = 2–4). weeks post-injection (WPI), cSLO imaging document eGFP EV2A) revealed all constant detector gain observe comparable levels fluorescence cSLO, it achieved brightest WPI. subsequent histological analysis cryosections 3 WPI EV2B) verified optimal since highest confirm decision, revised screened related sequences parallel. found GGSPPYR SLGGSPR closely EV2C), while might hint PTPS/LR receptor-binding motif EV2D). Table 1. Capsid identified screening. Abbreviation variant Enrichment factor rods/whole cones/whole GLSPPTR n/d 36 NNPTPSR 137 AAV2.GA GAHRSDS 28 NSRPAAA 4 SSPGLPR Sequence follows: RGN587 (AAV2 sequence)—AAA (linker sequence)—7mer insertion—AA sequence)—RQ sequence). enrichment calculated versus cones, corresponding population. EV2. Selection candidate Comparative packaged sc-CMV-eGFP; vg. A. reporting murine B. Confocal scans native 3WPI. Scale bar: 50 μm. Acquisition settings kept samples. GFP eGFP. C, D. Phylogenetic tree (C) (D) 3. peptide inserts aligned, phylogenetic constructed maximum likelihood method. Numbers bootstrap values, highlighted. compared AAV2.7m8 evaluate comprehensively performance AAV2.GL(GL) AAV2.NN(NN) AAV2.7m8(7m8) (Dalkara 2013) state-of-art reference. sc-CMV-eGFP cassette packaged, wild-type 3–4). up weekly fluorescence. Animals treated scattered weak 1 WPI, became gradually intense contrast, AAV2.7m8-, AAV2.GL- AAV2.NN-treated eyes strong expanded fundus 1A). euthanized, processed IHC. cross sections AAV2-treated signal mainly ganglion layer (GCL), sparse eGFP-positive inner (INL) nuclear (ONL). Engineered AAV2.7m8, however, spanned throughout 1B). Notably, stronger 1C) co-localised photoreceptor-specific recoverin 1D E). ≥ 3) 13.4-fold (P 0.0086) transcript 12.1-fold 0.0211) AAV2. While 5.5-fold transcripts, significantly different 0.2295) 1F). compare among capsids, previous 8E9 carrying stranded (ss) control rhodopsin (hRho) promoter injected. again monitored in-life week 2A). All outperformed AAV2, showing strongest signal. Specifically, faster brighter present rod-favouring nature 1). quantification confirmed finding showed higher level transcripts 0.0002), 0.0018) (0.0313). 0.0427) trending 0.2725). recorded AAV2.7m8-treated 0.2725) 2B). superiority ONL, least 2C). panretinal Vector mediated controls examinations 1- 2-week (WPI) (in µl) variants. immunolabelled DAPI. 100 C. tile-scans Hoechst. 500 D, E. Immunolabelled infected respectively, ONL INL, stained eGFP, F. qRT–PCR comparing fold change Biological replicates n Bars mean ± SEM. One-way ANOVA post hoc Holm–Sidak multiple comparisons test. *P ≤ 0.05, **P 0.01. Data information: eGFP; layer; layer. (F) Detailed statistical Appendix S1. available online [emmm202013392-sup-0003-SDataFig1.zip] 2. outperform 1-, 2- 3-week ss-hRho-eGFP relative ITR2 amplico

Language: Английский

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Environment shapes sleep patterns in a wild nocturnal primate

Scientific Reports, Journal Year: 2019, Volume and Issue: 9(1)

Published: July 9, 2019

Abstract Among primates, the suborder Haplorhini is considered to have evolved a consolidated monophasic sleep pattern, with diurnal species requiring shorter duration than nocturnal species. Only few primate been systematically studied in their natural habitat where environmental variables, including temperature and light, major influence on activity patterns. Here we report first study performed wild. We fitted seven wild Javan slow lorises ( Nycticebus javanicus ) West Java, Indonesia accelerometers that collected data, installed climate loggers each individual’s home range collect ambient readings (over 321 days total). All individuals showed strictly pattern of displayed striking synchronisation onset cessation relation sunset sunrise. The longest rest episodes were typically clustered near beginning towards end light period, this was inversely related daily fluctuations temperature. relationship between patterns, levels suggests role environment shaping architecture waking sleep. concluded well-known phenotypic variability amount across may represent an adaptation changes environment. Our data suggest patterns shaped by pressures observed phylogenetic inertia evolution humans.

Language: Английский

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