Heterogeneous Expression of Nuclear Encoded Mitochondrial Genes Distinguishes Inhibitory and Excitatory Neurons DOI Creative Commons
Meghan E. Wynne, Alicia R. Lane, Kaela S. Singleton

et al.

eNeuro, Journal Year: 2021, Volume and Issue: 8(4), P. ENEURO.0232 - 21.2021

Published: July 1, 2021

Mitochondrial composition varies by organ and their constituent cell types. This mitochondrial diversity likely determines variations in function. However, the heterogeneity of mitochondria brain remains underexplored despite large types neuronal tissue. Here, we used molecular systems biology tools to address whether region type mice. We reasoned that proteomics transcriptomics microdissected regions combined with analysis single-cell mRNA sequencing (scRNAseq) could reveal extent compositional diversity. selected nuclear encoded gene products forming complexes fixed stoichiometry, such as respiratory chain ribosome, well molecules perform function monomers, family SLC25 transporters. found proteome encompassing these nuclear-encoded genes obtained from tissue segregated hippocampus, striatum, cortex each other. Nuclear-encoded transcripts only segregate when was performed at level. In fact, transcriptomes were able distinguish glutamatergic distinct GABAergic neurons one another. Within categories, unique SLC25A transporters identify subpopulations. Our results demonstrate heterogeneous across postulate influences regional type-specific mechanisms health disease.

Language: Английский

A perspective on astrocyte regulation of neural circuit function and animal behavior DOI
Johannes Hirrlinger, Axel Nimmerjahn

Glia, Journal Year: 2022, Volume and Issue: 70(8), P. 1554 - 1580

Published: March 17, 2022

Studies over the past two decades have demonstrated that astrocytes are tightly associated with neurons and play pivotal roles in neural circuit development, operation, adaptation health disease. Nevertheless, precisely how integrate diverse neuronal signals, modulate structure function at multiple temporal spatial scales, influence animal behavior or disease through aberrant excitation molecular output remains unclear. This Perspective discusses new state-of-the-art approaches, including fluorescence indicators, opto- chemogenetic actuators, genetic targeting tools, quantitative behavioral assays, computational methods, might help resolve these longstanding questions. It also addresses complicating factors interpreting astrocytes' role regulation behavior, such as their heterogeneity, metabolism, inter-glial communication. Research on questions should provide a deeper mechanistic understanding of astrocyte-neuron assemblies' function, complex behaviors,

Language: Английский

Citations

38
Neurophotonic Tools for Microscopic Measurements and Manipulation: Status Report DOI Creative Commons
Ahmed S. Abdelfattah,

Srinivasa Rao Allu,

Robert E. Campbell

et al.

Neurophotonics, Journal Year: 2022, Volume and Issue: 9(S1)

Published: April 27, 2022

Neurophotonics was launched in 2014 coinciding with the launch of BRAIN Initiative focused on development technologies for advancement neuroscience. For last seven years, Neurophotonics' agenda has been well aligned this focus neurotechnologies featuring new optical methods and tools applicable to brain studies. While 2.0 is pivoting towards applications these novel quest understand brain, article we review an extensive diverse toolkit explore function that have emerged from related large-scale efforts measurement manipulation structure function. Here, neurophotonic mostly animal A companion article, scheduled appear later year, will cover diffuse imaging noninvasive human each domain, outline current state-of-the-art respective technologies, identify areas where innovation needed provide outlook future directions.

Language: Английский

Citations

33
Fluorogenic Rhodamine-Based Chemigenetic Biosensor for Monitoring Cellular NADPH Dynamics DOI

Huimin Chang,

Simon Clemens,

Pingting Gao

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(30), P. 20569 - 20576

Published: July 22, 2024

Ratiometric biosensors employing Förster Resonance Energy Transfer (FRET) enable the real-time tracking of metabolite dynamics. Here, we introduce an approach for generating a FRET-based biosensor in which changes apparent FRET efficiency rely on analyte-controlled fluorogenicity rhodamine rather than commonly used distance change between donor-acceptor fluorophores. Our fluorogenic, rhodamine-based, chemigenetic (

Language: Английский

Citations

9
State-dependent motion of a genetically encoded fluorescent biosensor DOI Creative Commons
Paul C. Rosen, Samantha M. Horwitz, Daniel J. Brooks

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(10)

Published: March 6, 2025

Genetically encoded biosensors can measure biochemical properties such as small-molecule concentrations with single-cell resolution, even in vivo. Despite their utility, these sensors are "black boxes": Very little is known about the structures of low- and high-fluorescence states or what features required to transition between them. We used LiLac, a lactate biosensor quantitative fluorescence-lifetime readout, model system address questions. X-ray crystal engineered high-affinity metal bridges demonstrate that LiLac exhibits large interdomain twist motion pulls fluorescent protein away from "sealed," high-lifetime state absence "cracked," low-lifetime its presence. Understanding dynamics will help think engineer other biosensors.

Language: Английский

Citations

1
Developmental energetics: Energy expenditure, budgets and metabolism during animal embryogenesis DOI Creative Commons
Suhrid Ghosh,

Anna Körte,

Giulia Serafini

et al.

Seminars in Cell and Developmental Biology, Journal Year: 2022, Volume and Issue: 138, P. 83 - 93

Published: March 19, 2022

Developing embryos are metabolically active, open systems that constantly exchange matter and energy with their environment. They function out of thermodynamic equilibrium continuously use metabolic pathways to obtain from maternal nutrients, in order fulfill the energetic requirements growth development. While an increasing number studies highlight role metabolism different developmental contexts, physicochemical basis embryogenesis, or how cellular processes act together transform a zygote into adult organism, remains unknown. As we better understanding metabolism, benefit current technology development, it is promising time revisit cost development principles may govern embryogenesis. Here, review recent advances methodology measure infer parameters developing embryos. We potential common pattern embryonic expenditure strategy across animal discuss challenges questions energetics.

Language: Английский

Citations

27
Genetically encoded fluorescent sensors for imaging neuronal dynamics in vivo DOI Open Access
Julian Day‐Cooney, Rochelin Dalangin, Haining Zhong

et al.

Journal of Neurochemistry, Journal Year: 2022, Volume and Issue: 164(3), P. 284 - 308

Published: March 14, 2022

The brain relies on many forms of dynamic activities in individual neurons, from synaptic transmission to electrical activity and intracellular signaling events. Monitoring these neuronal with high spatiotemporal resolution the context animal behavior is a necessary step achieve mechanistic understanding function. With rapid development dissemination highly optimized genetically encoded fluorescent sensors, growing number can now be visualized vivo. To date, cellular calcium imaging, which has been largely used as proxy for activity, become mainstay systems neuroscience. While challenges remain, voltage imaging neural populations possible. In addition, it becoming increasingly practical image over half dozen neurotransmitters, well certain metabolic activities. These new capabilities enable neuroscientists test previously unattainable hypotheses questions. This review summarizes recent progress delivery highlights example applications vivo imaging.

Language: Английский

Citations

27
Deficits in mitochondrial TCA cycle and OXPHOS precede rod photoreceptor degeneration during chronic HIF activation DOI Creative Commons
Vyara Todorova,

Mia Fee Stauffacher,

Luca Ravotto

et al.

Molecular Neurodegeneration, Journal Year: 2023, Volume and Issue: 18(1)

Published: March 7, 2023

Abstract Background Major retinal degenerative diseases, including age-related macular degeneration, diabetic retinopathy and detachment, are associated with a local decrease in oxygen availability causing the formation of hypoxic areas affecting photoreceptor (PR) cells. Here, we addressed underlying pathological mechanisms PR degeneration by focusing on energy metabolism during chronic activation hypoxia-inducible factors (HIFs) rod PR. Methods We used two-photon laser scanning microscopy (TPLSM) genetically encoded biosensors delivered adeno-associated viruses (AAV) to determine lactate glucose dynamics inner Retinal layer-specific proteomics, situ enzymatic assays immunofluorescence studies were analyse mitochondrial PRs HIF activation. Results exhibited remarkably higher glycolytic flux through hexokinases than neurons retina. Chronic rods did not cause overt change but an increase production nonetheless. Furthermore, dysregulation oxidative phosphorylation pathway (OXPHOS) tricarboxylic acid (TCA) cycle activated response decelerated cellular anabolism shortening outer segments (OS) before onset cell degeneration. Interestingly, deficient OXPHOS intact TCA exhibit these early signs anabolic showed slower course Conclusion Together, data indicate exceeding high highlight importance especially for survival conditions increased activity.

Language: Английский

Citations

14
Gray and white matter astrocytes differ in basal metabolism but respond similarly to neuronal activity DOI Creative Commons
Susanne Köhler,

Ulrike Winkler,

Tabea Junge

et al.

Glia, Journal Year: 2022, Volume and Issue: 71(2), P. 229 - 244

Published: Sept. 5, 2022

Abstract Astrocytes are a heterogeneous population of glial cells in the brain, which adapt their properties to requirements local environment. Two major groups astrocytes protoplasmic residing gray matter as well fibrous white matter. Here, we compared energy metabolism cortex and corpus callosum representative regions, acute brain slices taking advantage genetically encoded fluorescent nanosensors for NADH/NAD + redox ratio ATP. presented more reduced basal ratio, lower cytosolic concentration ATP cortical astrocytes. In astrocytes, neurotransmitter glutamate increased extracellular concentrations K , typical correlates neuronal activity, induced ratio. While application decreased [ATP], combination resulted an increase levels. Strikingly, very similar regulation by was observed callosum. Finally, strong intrinsic activity provoked bicuculline withdrawal Mg 2+ caused shift state slight reduction [ATP] summary, shows distinct properties, but qualitatively responses probably reflecting different environment these regions.

Language: Английский

Citations

22
Enlightening brain energy metabolism DOI Creative Commons
L. Felipe Barros, Iván Ruminot, Pamela Y. Sandoval

et al.

Neurobiology of Disease, Journal Year: 2023, Volume and Issue: 184, P. 106211 - 106211

Published: June 21, 2023

Brain tissue metabolism is distributed across several cell types and subcellular compartments, which activate at different times with temporal patterns. The introduction of genetically-encoded fluorescent indicators that are imaged using time-lapse microscopy has opened the possibility studying brain cellular sub-cellular levels. There for sugars, monocarboxylates, Krebs cycle intermediates, amino acids, cofactors, energy nucleotides, inform about relative levels, concentrations fluxes. This review offers a brief survey metabolic have been validated in cells, some illustrative examples from literature. Whereas only small fraction metabolome currently accessible to probes, there grounds be optimistic coming developments application these tools study disease.

Language: Английский

Citations

12
An engineered biosensor enables dynamic aspartate measurements in living cells DOI Creative Commons
Kristian Davidsen, Jonathan S. Marvin, Abhi Aggarwal

et al.

eLife, Journal Year: 2024, Volume and Issue: 12

Published: Feb. 23, 2024

Intracellular levels of the amino acid aspartate are responsive to changes in metabolism mammalian cells and can correspondingly alter cell function, highlighting need for robust tools measure abundance. However, comprehensive understanding has been limited by throughput, cost, static nature mass spectrometry (MS)-based measurements that typically employed levels. To address these issues, we have developed a green fluorescent protein (GFP)-based sensor (jAspSnFR3), where fluorescence intensity corresponds concentration. As purified protein, 20-fold increase upon saturation, with dose-dependent covering physiologically relevant concentration range no significant off target binding. Expressed lines, correlated measured MS could resolve temporal intracellular from genetic, pharmacological, nutritional manipulations. These data demonstrate utility jAspSnFR3 highlight opportunities it provides temporally resolved high-throughput applications variables affect

Language: Английский

Citations

5