Eddy, N. | bioRxiv Science

bioRxiv2024

Mitochondrial oxidation of the carbohydrate fuel driven by pyruvate dehydrogenase robustly enhances stemness of older and geriatric Intestinal Stem Cells

Background and AimsAging impairs Intestinal Stem Cell (ISC) function and attenuates their regenerative capacity. Although the transcriptional landscape governing ISC fate during aging has been described, almost nothing is known about how metabolite handling regulates ISC renewal and maintains stemness. We investigated how mitochondrial metabolism of glucose and fatty acid-derived carbons, regulated by the gatekeeper, pyruvate dehydrogenase (PDH) rescues ISC stemness in older and geriatric mice and humans. MethodsProximal small intestinal organoids (enteroids) generated from pinch biopsy specimens obtained from young (21-25y) and older individuals (64-75y), and GFP-sorted single ISCs from Lgr5-EGFP mice (2-24 months) were used to examine hallmarks of ISC stemness. Mitochondrial morphology was evaluated using transmission electron microscopy. Mitochondrial oxygen consumption rate (OCR), ATP (mitoATP), and glycolytic ATP production were measured in the presence of full and single metabolic substrates (pyruvate, glutamate, and fatty acids) in whole cell and isolated mitochondria using the high throughput Seahorse XF technology. Carbon flux through TCA cycle was determined by 13C6-glucose tracing and measuring 13C enrichment in TCA cycle intermediates using liquid chromatography mass spectrometry. ResultsAge induced decline in ISC stemness is driven by a dramatic decrease in PDH activity that shuttles pyruvate away from the TCA cycle. Restoring PDH activity by inhibition of pyruvate dehydrogenase kinase 4 (PDK4) drives glucose-derived carbon entry into TCA cycle and subsequently increases mitochondrial OCR and mitoATP, collectively rescuing the decline in stemness in aging ISCs. The observed shift in fuel preference from fatty acids to glucose is unaltered by PDK4 inhibition. ConclusionPDH upregulation rescues age-induced decline in ISC stemness in humans and mice via directing glucose derived carbons to TCA cycle and increasing mitoATP production.

cell biology↗

bioRxiv2022

Kelpwatch: A new visualization and analysis tool to explore kelp canopy dynamics reveals variable resistance and resilience to marine heat waves

Giant kelp and bull kelp forests are increasingly at risk from marine heatwave events, herbivore outbreaks, and the loss or alterations in the behavior of key herbivore predators. The dynamic floating canopy of these kelps is well-suited to study via satellite imagery, which provides high temporal and spatial resolution data of floating kelp canopy across the western United States and Mexico. However, the size and complexity of the satellite image dataset has made ecological analysis difficult for scientists and managers. To increase accessibility of this rich dataset, we created Kelpwatch, a web-based visualization and analysis tool. This tool allows researchers and managers to quantify kelp forest change in response to disturbances, assess historical trends, and allow for effective and actionable kelp forest management. Here, we demonstrate how Kelpwatch can be used to analyze long-term trends in kelp canopy across regions, quantify spatial variability in the response to and recovery from the 2014 to 2016 marine heatwave events, and provide a local analysis of kelp canopy status around the Monterey Peninsula, California. We found that 18.6% of regional sites displayed a significant trend in kelp canopy area over the past 38 years and that there was a latitudinal response to heatwave events for each kelp species. The recovery from heatwave events was more variable across space, with some local areas like Bahia Tortugas in Baja California Sur showing high recovery while kelp canopies around the Monterey Peninsula continued a slow decline and patchy recovery compared to the rest of the Central California region. Kelpwatch provides near real time spatial data and analysis support and makes complex earth observation data actionable for scientists and managers, which can help identify areas for research, monitoring, and management efforts.

ecology↗