Mouse tissue harvest-induced hypoxia rapidly alters the in vivo metabolome, between-genotype metabolite level differences, and 13C-tracing enrichments

Rauckhorst, Adam J.; Borcherding, Nicholas; Pape, Daniel J.; Kraus, Alora S.; Scerbo, Diego; Taylor, Eric B.

doi:10.1016/j.molmet.2022.101596

Cited by 22 publications

(11 citation statements)

References 36 publications

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“…The relative abundances of 49 metabolites were significantly different in MPC TubKO kidneys ( Figure S4A, Supplemental Table 3 ). While pyruvate was not significantly increased, lactate and alanine, which are produced from pyruvate by single metabolic reactions and were previously identified to be increased in models of MPC disruption (28, 29, 37), were significantly increased in MPC TubKO kidneys ( Figure 4A ). TCA cycle metabolites were broadly decreased except for α-ketoglutarate, which is consistent with decreased pyruvate and increased glutamine oxidation ( Figure 4B ).…”

Section: Resultsmentioning

confidence: 73%

“…Whole kidneys were rapidly dissected from live mice under isoflurane anesthesia and then within 1-2 seconds freeze clamped with liquid nitrogen temperature tongs. Kidneys were extracted for metabolomics analysis (supplemental methods) by gas chromatography (GC)- and liquid chromatography (LC)-mass spectrometry (MS) as previously described (37). Acquired MS data were processed by Thermo Scientific TraceFinder 4.1 software.…”

Section: Methodsmentioning

confidence: 99%

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Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

Rauckhorst

Martinez

Andrade

et al. 2023

Preprint

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Energy-intensive kidney reabsorption processes essential for normal whole-body function are maintained by tubular epithelial cell metabolism. Tubular metabolism changes markedly following acute kidney injury (AKI), but which changes are adaptive versus maladaptive remain poorly understood. In publicly available data sets, we noticed a consistent downregulation of the mitochondrial pyruvate carrier (MPC) after AKI, which we experimentally confirmed. To test the functional consequences of MPC downregulation, we generated novel tubular epithelial cell-specific Mpc1 knockout (MPC TubKO) mice.13C-glucose tracing, steady-state metabolomic profiling, and enzymatic activity assays revealed that MPC TubKO coordinately increased activities of the pentose phosphate pathway and the glutathione and thioredoxin oxidant defense systems. Following rhabdomyolysis-induced AKI, MPC TubKO decreased markers of kidney injury and oxidative damage and strikingly increased survival. Our findings suggest that decreased mitochondrial pyruvate uptake is a central adaptive response following AKI and raise the possibility of therapeutically modulating the MPC to attenuate AKI severity.

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Section: Resultsmentioning

confidence: 73%

Section: Methodsmentioning

confidence: 99%

Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

Rauckhorst

Martinez

Andrade

et al. 2023

Preprint

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“…In this study, we aimed to improve the assessment of the brain metabolome by decreasing the post-mortem impact following brain resection. We would like to highlight that rodent brain metabolism is highly sensitive to stimuli such as handling and stress 72 , anesthesia 73 , and time post-euthanasia 29 . Additional steps will be needed to control these factors in the future to truly capture the “normal” brain metabolome.…”

Section: Discussionmentioning

confidence: 99%

“…Cellular metabolism continues post-mortem in rodents with signs of hypoxia driven metabolic reprogramming 29 . These metabolic changes are especially true for the brain, and capturing an accurate snapshot of the brain metabolome remains a critical challenge in the field.…”

Section: In Situ Microwave Fixation To Define Brain Metabolismmentioning

confidence: 99%

“…Post-mortem surgical removal of the brain ranges from 90 seconds to several minutes depending on the individual performing the resection. It is well documented that metabolism changes post-mortem 28,29 , and the rates of metabolic flux through glycolysis and the TCA cycle occurs in seconds [30][31][32] . Furthermore, artificial tissue hypoxia has been reported in mice following cervical dislocation 33 , decapitation 34 , and CO2 euthanasia 35 .…”

Section: Introductionmentioning

confidence: 99%

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In Situ Microwave Fixation to Define the Terminal Rodent Brain Metabolome

Juras

Webb

Young

et al. 2022

Preprint

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The brain metabolome directly connects to brain physiology and neuronal function. Brain glucose metabolism is highly heterogeneous among brain regions and continues postmortem. Therefore, challenges remain to capture an accurate snapshot of the physiological brain metabolome in healthy and diseased rodent models. To overcome this barrier, we employ a high-power focused microwave for the simultaneous euthanasia and fixation of mouse brain tissue to preserve metabolite pools prior to surgical removal and dissection of brain regions. We demonstrate exhaustion of glycogen and glucose and increase in lactate production during conventional rapid brain resection prior to preservation by liquid nitrogen that is not observed with microwave fixation. Next, microwave fixation was employed to define the impact of brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple regions of the mouse brain, evidenced by reduced 13C enrichment into glycogen, glycolysis, and the TCA cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation to study terminal brain metabolism in rodent models.

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Perspectives on the Drosophila melanogaster Model for Advances in Toxicological Science

Rand,

Tennessen,

Mackay

et al. 2023

Current Protocols

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The use of Drosophila melanogaster for studies of toxicology has grown considerably in the last decade. The Drosophila model has long been appreciated as a versatile and powerful model for developmental biology and genetics because of its ease of handling, short life cycle, low cost of maintenance, molecular genetic accessibility, and availability of a wide range of publicly available strains and data resources. These features, together with recent unique developments in genomics and metabolomics, make the fly model especially relevant and timely for the development of new approach methodologies and movements toward precision toxicology. Here, we offer a perspective on how flies can be leveraged to identify risk factors relevant to environmental exposures and human health. First, we review and discuss fundamental toxicologic principles for experimental design with Drosophila. Next, we describe quantitative and systems genetics approaches to resolve the genetic architecture and candidate pathways controlling susceptibility to toxicants. Finally, we summarize the current state and future promise of the emerging field of Drosophila metabolomics for elaborating toxic mechanisms. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.

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Mouse tissue harvest-induced hypoxia rapidly alters the in vivo metabolome, between-genotype metabolite level differences, and 13C-tracing enrichments

Cited by 22 publications

References 36 publications

Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

In Situ Microwave Fixation to Define the Terminal Rodent Brain Metabolome

Perspectives on the Drosophila melanogaster Model for Advances in Toxicological Science

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