The plasma membrane, which is a phosphoglyceride bilayer at the outer edge of the cell, plays diverse and important roles in biological systems. Visualization of the plasma membrane in live samples is important for various applications in biological functions. We developed an amphiphilic two-photon (TP) fluorescent probe (THQ-Mem) to selectively monitor the plasma membrane in live samples. This probe exhibited red emission (620-700 nm), large TP absorption cross sections (δmax > 790 GM), and high selectivity to the plasma membrane. In cultured cells and in vivo hepatic tissue imaging, THQ-Mem showed bright TP-excited fluorescence (TPEF) and remarkable selectivity for the plasma membrane. Furthermore, simultaneous in vivo imaging with THQ-Mem and a TP lipid droplet probe could serve as an efficient tool to monitor morphological and physiological changes in the plasma membrane and lipid droplets.

In Vivo Simultaneous Imaging of Plasma Membrane and Lipid Droplets in Hepatic Steatosis using Red-Emissive Two-Photon Probes.


 Gestational hyperandrogenism adversely impacts offspring health. Using an ovine model, we found prenatal testosterone (T) excess adversely affects growth and cardiometabolic outcomes in female offspring and sex-specific effects on fetal myocardium. Since lipids are essential to cardiometabolic function, we hypothesized that prenatal T excess will lead to sex-specific disruptions in lipid metabolism at birth. Shotgun lipidomics was performed on the plasma samples collected 48 hours after birth from female (F) and male (M) lambs of control (C) and (T) sheep (CF=4, TF=7, CM=5, TM=10) and data analyzed by univariate analysis, multivariate dimensionality reduction modeling followed by functional enrichment and pathway analyses. Biosynthesis of phosphatidylserine was the major pathway responsible for sex differences in controls. Unsupervised and supervised models showed separation between C and T in both sexes with glycerophospholipids and glycerolipids classes being responsible for the sex differences between C and T. T excess increased cholesterol in females while decreasing phosphatidylcholine levels in male lambs. Specifically, T excess: 1) suppressed phosphatidylethanolamine N-methyltransferase (PEMT) phosphatidylcholine synthesis pathway overall and in TM lambs as opposed to suppression of carnitine levels overall and TF lambs and 2) activated biosynthesis of PE-O and PC-O from DG-O overall and in TF lambs. Higher cholesterol levels could underlie adverse cardiometabolic outcomes in TF lambs, whereas suppressed PEMT pathway in TM lambs could lead to endoplasmic reticulum stress and defect in lipid transport. These novel findings point to sex-specific effects of prenatal T excess on lipid metabolism of newborn lamb, a precocial ovine model of translational relevance.

Sex-specific perturbation of systemic lipidomic profile in newborn lambs impacted by prenatal testosterone excess

Delivery of Eicosapentaenoic acid-loaded cellulose nanocrystal alleviates liver fibrosis via modulating phospholipids and apoptotic regulators

Alcohol consumption and high-fat diets often coincide in Western society, exerting negative synergistic effects on the liver. While many studies have demonstrated the impact of ALD and NAFLD on organ protein expression, none have offered a comprehensive view of the dysregulation at the level of the membrane proteome. In this study, we utilize peptidisc and solvent precipitation (SP4) methods to isolate and compare the membrane protein content of the liver with its unique biological functions. Using mice treated with a high-fat diet and ethanol in drinking water, we identified 1,563 liver proteins, with 46% predicted to have a transmembrane segment. Among these, 106 integral membrane proteins are dysregulated compared to the untreated sample. Gene ontology analysis reveals several dysregulated membrane processes associated with lipid metabolism, cell adhesion, xenobiotic processing, and mitochondrial membrane formation. Pathways related to cholesterol and bile acid transport are also mutually affected, suggesting an adaptive mechanism to counter the steatosis of the liver model. Our peptidisc-based membrane proteome profiling thus emerges as an effective way to gain insights into the role of the transmembrane proteome in disease development, warranting further in-depth analysis of the individual effect of the identified dysregulated membrane proteins.

Profiling the organ membrane proteome dysregulation in the context of liver disease

Alcohol consumption and high-fat diets often coincide in Western society, exerting negative synergistic effects on the liver. While many studies have demonstrated the impact of ALD and NAFLD on organ protein expression, none have offered a comprehensive view of the dysregulation at the level of the membrane proteome. In this study, we utilize peptidisc and solvent precipitation (SP4) methods to isolate and compare the membrane protein content of the liver with its unique biological functions. Using mice treated with a high-fat diet and ethanol in drinking water, we identified 1,563 liver proteins, with 46% predicted to have a transmembrane segment. Among these, 106 integral membrane proteins are dysregulated compared to the untreated sample. Gene ontology analysis reveals several dysregulated membrane processes associated with lipid metabolism, cell adhesion, xenobiotic processing, and mitochondrial membrane formation. Pathways related to cholesterol and bile acid transport are also mutually affected, suggesting an adaptive mechanism to counter the steatosis of the liver model. Our peptidisc-based membrane proteome profiling thus emerges as an effective way to gain insights into the role of the transmembrane proteome in disease development, warranting further in-depth analysis of the individual effect of the identified dysregulated membrane proteins. 

Obesity has reached global epidemic proportions and it affects the development of insulin resistance, type 2 diabetes, fatty liver disease and other metabolic diseases. Membrane lipids are important structural and signaling components of the cell membrane. Recent studies highlight their importance in lipid homeostasis and are implicated in the pathogenesis of fatty liver disease. Here, we discuss the numerous membrane lipid species and their metabolites including, phospholipids, sphingolipids and cholesterol, and how dysregulation of their composition and physiology contribute to the development of fatty liver disease. The development of new genetic and pharmacological mouse models has shed light on the role of lipid species on various mechanisms/pathways; these lipids impact many aspects of the pathophysiology of fatty liver disease and could potentially be targeted for the treatment of fatty liver disease.

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Characterization and Roles of Membrane Lipids in Fatty Liver Disease

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Characterization and Roles of Membrane Lipids in Fatty Liver Disease