Intravital two-photon imaging and quantification of hepatic steatosis and fibrosis in a live small animal model

Moon, Jaekyun; Jeon, Jehwi; Kong, Eunji; Hong, Sujung; Lee, Jingu; Lee, Eun Kyung; Kim, Pilhan

doi:10.1364/boe.442608

Cited by 12 publications

(5 citation statements)

References 49 publications

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“…NAFLD can progress to nonalcoholic steatohepatitis (NASH) via inflammation and fibrosis [ 27 ]. Bio-imaging data suggest that, as fatty liver progresses in mice, tiny individual hepatic LDs interact and fuse to form large LDs [ 28 ]. During the fatty liver progression, the area and length of sinusoidal vessels are decreased [ 14 ] and nonparenchymal cells, including infiltrated leucocytes, are pathologically changed [ 15 ].…”

Section: Discussionmentioning

confidence: 99%

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In Vivo Two-Photon Imaging Analysis of Dynamic Degradation of Hepatic Lipid Droplets in MS-275-Treated Mouse Liver

Lee

Park

et al. 2022

IJMS

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The accumulation of hepatic lipid droplets (LDs) is a hallmark of non-alcoholic fatty liver disease (NAFLD). Appropriate degradation of hepatic LDs and oxidation of complete free fatty acids (FFAs) are important for preventing the development of NAFLD. Histone deacetylase (HDAC) is involved in the impaired lipid metabolism seen in high-fat diet (HFD)-induced obese mice. Here, we evaluated the effect of MS-275, an inhibitor of HDAC1/3, on the degradation of hepatic LDs and FFA oxidation in HFD-induced NAFLD mice. To assess the dynamic degradation of hepatic LDs and FFA oxidation in fatty livers of MS-275-treated HFD C57BL/6J mice, an intravital two-photon imaging system was used and biochemical analysis was performed. The MS-275 improved hepatic metabolic alterations in HFD-induced fatty liver by increasing the dynamic degradation of hepatic LDs and the interaction between LDs and lysozyme in the fatty liver. Numerous peri-droplet mitochondria, lipolysis, and lipophagy were observed in the MS-275-treated mouse fatty liver. Biochemical analysis revealed that the lipolysis and autophagy pathways were activated in MS-275 treated mouse liver. In addition, MS-275 reduced the de novo lipogenesis, but increased the mitochondrial oxidation and the expression levels of oxidation-related genes, such as PPARa, MCAD, CPT1b, and FGF21. Taken together, these results suggest that MS-275 stimulates the degradation of hepatic LDs and mitochondrial free fatty acid oxidation, thus protecting against HFD-induced NAFLD.

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Section: Discussionmentioning

confidence: 99%

“…ZEN 3.2 (Carl Zeiss), Volocity (Quorum Technologies Inc., Puslinch, ON, Canada), and Imaris 9.3.1 (Bitplane, Zurich, Switzerland) software were used for the 3D image analysis. The motion and volume of hepatic LDs and lysosomes were quantified and visualized using the surface function of Imaris, as previously described [ 28 ].…”

Section: Methodsmentioning

confidence: 99%

In Vivo Two-Photon Imaging Analysis of Dynamic Degradation of Hepatic Lipid Droplets in MS-275-Treated Mouse Liver

Lee

Park

et al. 2022

IJMS

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“…The exposed kidney surface was covered by transparent coverslip 52 , 53 . A wet gauze soaked in warm saline was placed between the kidney and the underlying tissue to reduce motion artifacts 54 – 56 . To image muscle, a 10 mm incision was made on thigh skin and then muscle was exposed and covered by transparent coverslip 57 .…”

Section: Methodsmentioning

confidence: 99%

Statistically unbiased prediction enables accurate denoising of voltage imaging data

Eom,

Han,

Park

et al. 2023

Nat Methods

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Here we report SUPPORT (statistically unbiased prediction utilizing spatiotemporal information in imaging data), a self-supervised learning method for removing Poisson–Gaussian noise in voltage imaging data. SUPPORT is based on the insight that a pixel value in voltage imaging data is highly dependent on its spatiotemporal neighboring pixels, even when its temporally adjacent frames alone do not provide useful information for statistical prediction. Such dependency is captured and used by a convolutional neural network with a spatiotemporal blind spot to accurately denoise voltage imaging data in which the existence of the action potential in a time frame cannot be inferred by the information in other frames. Through simulations and experiments, we show that SUPPORT enables precise denoising of voltage imaging data and other types of microscopy image while preserving the underlying dynamics within the scene.

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“…Imaging using this optimized IVM protocol provided details of cell trafficking, recruitment, function, and behavior in fatty liver ( Davis et al, 2019 ). Moon et al used intravital two-photon microscopy to quantitate lipid droplet accumulation, fibrosis, and disruption of the microvasculature ( Moon et al, 2020 ; Moon et al, 2021 ). These methods enable studying the inflammatory response and disease progression in NAFLD and can facilitate the development of diagnostic and therapeutic strategies.…”

Section: Novel Ivm Methods To Investigate Dynamic Single-cell Behaviormentioning

confidence: 99%

Intravital Imaging of Inflammatory Response in Liver Disease

Wang

Wang²

2022

Front. Cell Dev. Biol.

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The healthy liver requires a strictly controlled crosstalk between immune and nonimmune cells to maintain its function and homeostasis. A well-conditioned immune system can effectively recognize and clear noxious stimuli by a self-limited, small-scale inflammatory response. This regulated inflammatory process enables the liver to cope with daily microbial exposure and metabolic stress, which is beneficial for hepatic self-renewal and tissue remodeling. However, the failure to clear noxious stimuli or dysregulation of immune response can lead to uncontrolled liver inflammation, liver dysfunction, and severe liver disease. Numerous highly dynamic circulating immune cells and sessile resident immune and parenchymal cells interact and communicate with each other in an incredibly complex way to regulate the inflammatory response in both healthy and diseased liver. Intravital imaging is a powerful tool to visualize individual cells in vivo and has been widely used for dissecting the behavior and interactions between various cell types in the complex architecture of the liver. Here, we summarize some new findings obtained with the use of intravital imaging, which enhances our understanding of the complexity of immune cell behavior, cell–cell interaction, and spatial organization during the physiological and pathological liver inflammatory response.

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Intravital two-photon imaging and quantification of hepatic steatosis and fibrosis in a live small animal model

Cited by 12 publications

References 49 publications

In Vivo Two-Photon Imaging Analysis of Dynamic Degradation of Hepatic Lipid Droplets in MS-275-Treated Mouse Liver

In Vivo Two-Photon Imaging Analysis of Dynamic Degradation of Hepatic Lipid Droplets in MS-275-Treated Mouse Liver

Statistically unbiased prediction enables accurate denoising of voltage imaging data

Intravital Imaging of Inflammatory Response in Liver Disease

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