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Laser microdissection-based microproteomics of the hippocampus of a rat epilepsy model reveals regional differences in protein abundances
摘要： Mesial temporal lobe epilepsy (MTLE) is a chronic neurological disorder affecting almost 40% of adult patients with epilepsy. Hippocampal sclerosis (HS) is a common histopathological abnormality found in patients with MTLE. HS is characterised by extensive neuronal loss in different hippocampus sub-regions. In this study, we used laser microdissection-based microproteomics to determine the protein abundances in different regions and layers of the hippocampus dentate gyrus (DG) in an electric stimulation rodent model which displays classical HS damage similar to that found in patients with MTLE. Our results indicate that there are differences in the proteomic profiles of different layers (granule cell and molecular), as well as different regions, of the DG (ventral and dorsal). We have identified new signalling pathways and proteins present in specific layers and regions of the DG, such as PARK7, RACK1, and connexin 31/gap junction. We also found two major signalling pathways that are common to all layers and regions: inflammation and energy metabolism. Finally, our results highlight the utility of high-throughput microproteomics and spatial-limited isolation of tissues in the study of complex disorders to fully appreciate the large biological heterogeneity present in different cell populations within the central nervous system.
关键词： microproteomics,rat epilepsy model,hippocampus,protein abundances,Laser microdissection,PARK7,energy metabolism,connexin 31/gap junction,RACK1,inflammation
更新于2025-09-19 17:13:59
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Fluorescence Lifetime Imaging Ophthalmoscopy of the Retinal Pigment Epithelium During Wound Healing After Laser Irradiation
摘要： Purpose: To investigate the change in fluorescence lifetime of retinal pigment epithelium (RPE) after laser irradiation by using an organ culture model. Methods: Porcine RPE-choroid-sclera explants were irradiated with selective retina treatment laser (wavelength: 527 nm, beam diameter: 200 lm, energy: 80–150 lJ). At 24 and 72 hours after irradiation, the mean fluorescence lifetime (sm) was measured with fluorescence lifetime imaging ophthalmoscopy (FLIO) (excitation wavelength: 473 nm, emission: short spectral channel: 498-560 nm, long spectral channel: 560–720 nm). For every laser spot, central damaged zone (zone 1: 120 3 120 lm), area including wound rim (280 3 280 lm except zone 1), and environmental zone (440 3 440 lm except zone 1 and 2) were analyzed. Peripheral zone at a distance from laser spots longer than 2000 lm was examined for comparison. Cell viability was evaluated with calcein-acetoxymethyl ester and morphology with fluorescence microscopy for filamentous-actin. Results: The RPE defect after selective retina treatment was mostly closed within 72 hours. FLIO clearly demarcated the irradiated region, with prolonged sm at the center of the defect decreasing with eccentricity. In short spectral channel, but not in long spectral channel, sm in the environmental zone after 72 hours was still significantly longer than in the peripheral zone. Conclusions: FLIO may clearly demarcate the RPE defect, demonstrate its closure, and, moreover, indicate the induced metabolic changes of surrounding cells during wound healing. Translational Relevance: This ex vivo study showed that FLIO may be used to evaluate the extent and quality of restoration of the damaged RPE and to detect its metabolic change in human fundus noninvasively.
关键词： selective retina therapy,retinal pigment epithelium,fluorescence lifetime,energy metabolism,wound healing
更新于2025-09-16 10:30:52
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Anti-Myocardial Infarction Effects of Radix Aconiti Lateralis Preparata Extracts and Their Influence on Small Molecules in the Heart Using Matrix-Assisted Laser Desorption/Ionization–Mass Spectrometry Imaging
摘要： Radix Aconiti Lateralis Preparata (fuzi) is the processed product of Aconitum carmichaelii Debeaux tuber, and has great potential anti-myocardial infarction e?ects, including improving myocardial damage and energy metabolism in rats. However, the e?ects of Radix Aconiti Lateralis Preparata extracts in a rat model of myocardial infarction have not yet been fully illustrated. Herein, Radix Aconiti Lateral Preparata was used to prepare Radix Aconiti Lateralis Preparata extract (RAE), fuzi polysaccharides (FPS), and fuzi total alkaloid (FTA). Then, we aimed to compare the e?ects of RAE, FPS, and FTA in MI rats and further explore their in?uence on small molecules in the heart. We reported that Radix Aconiti Lateralis Preparata extract (RAE) and fuzi total alkaloid (FTA) signi?cantly improved left ventricular function and structure, and reduced myocardial damage and infarct size in rats with myocardial infarction by the left anterior descending artery ligation. In contrast, fuzi polysaccharides (FPS) was less e?ective than RAE and FTA, indicating that alkaloids might play a major role in the treatment of myocardial infarction. Moreover, via matrix-assisted laser desorption/ionization–mass spectrometry imaging (MALDI–MSI), we further showed that RAE and FTA containing alkaloids as the main common components regulated myocardial energy metabolism-related molecules and phospholipids levels and distribution patterns against myocardial infarction. In particular, it was FTA, not RAE, that could also regulate potassium ions and glutamine to play a cardioprotective role in myocardial infarction, which revealed that an appropriate dose of alkaloids generated more obvious cardiotonic e?ects. These ?ndings together suggested that Radix Aconiti Lateralis Preparata extracts containing an appropriate dose of alkaloids as its main pharmacological active components exerted protective e?ects against myocardial infarction by improving myocardial energy metabolism abnormalities and changing phospholipids levels and distribution patterns to stabilize the cardiomyocyte membrane structure. Thus, RAE and FTA extracted from Radix Aconiti Lateralis Preparata are potential candidates for the treatment of myocardial infarction.
关键词： myocardial infarction,left anterior descending artery ligation,phospholipid,energy metabolism,matrix-assisted laser desorption/ionization–mass spectrometry imaging,Radix Aconiti Lateralis Preparata
更新于2025-09-16 10:30:52
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ROLE OF CYCLIC AND PSEUDO-CYCLIC ELECTRON TRANSPORT IN RESPONSE TO DYNAMIC LIGHT CHANGES IN PHYSCOMITRELLA PATENS
摘要： Photosynthetic organisms support cell metabolism by harvesting sunlight and driving the electron transport chain at the level of thylakoid membranes. Excitation energy and electron flow in the photosynthetic apparatus is continuously modulated in response to dynamic environmental conditions. Alternative electron flow around photosystem I plays a seminal role in this regulation contributing to photo-protection by mitigating over-reduction of the electron carriers. Different pathways of alternative electron flow coexist in the moss Physcomitrella patens, including cyclic electron flow mediated by the PGRL1/PGR5 complex and pseudo-cyclic electron flow mediated by the flavodiiron proteins FLV. In this work we generated P. patens plants carrying both pgrl1 and flva knock-out (KO) mutations. A comparative analysis of the WT, pgrl1, flva and pgrl1 flva lines suggests that cyclic and pseudo-cyclic processes have a synergic role in the regulation of photosynthetic electron transport. However, while both contribute to photosystem I protection from over-reduction by modulating electron flow following changes in environmental conditions, FLV activity is particularly relevant in the first seconds after a light change while PGRL1 has a major role upon sustained strong illumination.
关键词： Photosynthetic Reaction Center Complex Proteins,Photosynthesis,photoprotection,Energy metabolism,Evolution, Molecular,Bryophyta
更新于2025-09-10 09:29:36