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

и другие.

eNeuro, Год журнала: 2021, Номер 8(4), С. ENEURO.0232 - 21.2021

Опубликована: Июль 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.

Язык: Английский

AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells DOI Creative Commons
Lars Hellweg, Martin Pfeifer, Mirosław Tarnawski

и другие.

Cell chemical biology, Год журнала: 2024, Номер 31(8), С. 1529 - 1541.e12

Опубликована: Май 27, 2024

Aspartate is crucial for nucleotide synthesis, ammonia detoxification, and maintaining redox balance via the malate-aspartate-shuttle (MAS). To disentangle these multiple roles of aspartate metabolism, tools are required that measure concentrations in real time live cells. We introduce AspSnFR, a genetically encoded green fluorescent biosensor intracellular aspartate, engineered through displaying screening libraries on mammalian In cells, AspSnFR able to precisely quantitatively cytosolic dissect its production from glutamine. Combining high-content imaging with pharmacological perturbations exposes differences metabolic vulnerabilities levels based nutrient availability. Further, facilitates tracking export mitochondria SLC25A12, MAS' key transporter. show SLC25A12 rapidly responding direct route couple Ca

Язык: Английский

Процитировано

5
Functional principles of genetically encoded fluorescent biosensors for metabolism and their quantitative use DOI Open Access
Dorothy Koveal

Journal of Neurochemistry, Год журнала: 2023, Номер 168(5), С. 496 - 505

Опубликована: Июнь 14, 2023

Genetically encoded fluorescent biosensors provide an attractive means of measuring chemical changes in single cells on fast timescales (milliseconds to seconds). While their most prominent application has been tracking neural activity and neurotransmitter release, there growing interest developing deploying new versions these tools study brain metabolism. However, the careful use interpretation data they remain challenging. Many are subject interferences that can alter sensor responses within a cell or between cells, producing ambiguous results. This presents challenge for quantitation our ability accurately interpret responses. review describes current methods quantitation, with focus cellular commonly affect performance, ways avoid false inferences, recent advances optimization make them more robust.

Язык: Английский

Процитировано

10
A ratiometric ER calcium sensor for quantitative comparisons across cell types and subcellular regions DOI Creative Commons
Ryan Farrell, Kirsten Bredvik, Michael B. Hoppa

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Фев. 15, 2024

Abstract The endoplasmic reticulum (ER) is an important regulator of Ca 2+ in cells and dysregulation ER calcium homeostasis can lead to numerous pathologies. Understanding how various pharmacological genetic perturbations impacts cellular physiology would likely be facilitated by more quantitative measurements levels that allow easier comparisons across conditions. Here, we developed a ratiometric version our original ER-GCaMP probe allows for the concentration cell types sub-cellular compartments. Using this approach show resting Ca2+ primary dissociated neurons substantially lower than measured embryonic fibroblasts.

Язык: Английский

Процитировано

4
Local glycolysis supports injury-induced axonal regeneration DOI Creative Commons
Luca Masin, Steven Bergmans, Annelies Van Dyck

и другие.

The Journal of Cell Biology, Год журнала: 2024, Номер 223(12)

Опубликована: Окт. 1, 2024

Successful axonal regeneration following injury requires the effective allocation of energy. How axons withstand initial disruption in mitochondrial energy production caused by and subsequently initiate regrowth is poorly understood. Transcriptomic data showed increased expression glycolytic genes after optic nerve crush retinal ganglion cells with co-deletion Pten Socs3. Using cultures a multicompartment microfluidic device, we observed enhanced trafficking Socs3 co-deleted neurons. While wild-type relied on metabolism, injury, absence Socs3, was supported local glycolysis. Specific inhibition lactate hindered survival initiation while slowing down glycolysis upstream impaired initiation, elongation, production. Together, these observations reveal that ATP, combined sustained transport, essential for injury-induced regrowth, providing new insights into metabolic underpinnings regeneration.

Язык: Английский

Процитировано

4
Heterogeneous Expression of Nuclear Encoded Mitochondrial Genes Distinguishes Inhibitory and Excitatory Neurons DOI Creative Commons
Meghan E. Wynne, Alicia R. Lane, Kaela S. Singleton

и другие.

eNeuro, Год журнала: 2021, Номер 8(4), С. ENEURO.0232 - 21.2021

Опубликована: Июль 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.

Язык: Английский

Процитировано

22