1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced Parkinson’s disease in zebrafish

Parkinson\u2019s disease (PD) is the most common age associated neurodegenerative disease, which has been extensively studied for its etiology and phenotype. PD has been widely studied in alternate model system such as rodents towards understanding the role of neurotoxin by inducing PD. This study is aimed to understand the biomechanism of PD in zebrafish model system induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The phenotype and role of various genes and proteins for Parkinsonism were tested and evaluated in this study using behaviour, molecular and proteomic approaches. Zebrafish PD model induced by 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyridine showed a significant level of decrease in the movement with erratic swimming pattern and increased freezing bouts. CHCHD2, EEF2B, LRRK2, PARK7, PARK2, POLG, SNCGB and SYNB genes were differentially regulated at the transcript level in PD zebrafish. Similarly a total of 73 proteins were recognized as differentially expressed in the nervous system of zebrafish due to Parkinsonism based on quantitative proteomics approach. Proteins such as NEFL,MUNC13-1, NAV2 and GAPVD1 were down regulated in the zebrafish brain for the PD phenotype, which were associated with the neurological pathways. This zebrafish based PD model can be used as a potential model system for screening prospective drug molecules for PD

Protein network analyses of pulmonary endothelial cells in chronic thromboembolic pulmonary hypertension

Chronic thromboembolic pulmonary hypertension (CTEPH) is a vascular disease characterized by the presence of organized thromboembolic material in pulmonary arteries leading to increased vascular resistance, heart failure and death. Dysfunction of endothelial cells is involved in CTEPH. The present study describes for the first time the molecular processes underlying endothelial dysfunction in the development of the CTEPH. The advanced analytical approach and the protein network analyses of patient derived CTEPH endothelial cells allowed the quantitation of 3258 proteins. The 673 differentially regulated proteins were associated with functional and disease protein network modules. The protein network analyses resulted in the characterization of dysregulated pathways associated with endothelial dysfunction, such as mitochondrial dysfunction, oxidative phosphorylation, sirtuin signaling, inflammatory response, oxidative stress and fatty acid metabolism related pathways. In addition, the quantification of advanced oxidation protein products, total protein carbonyl content, and intracellular reactive oxygen species resulted increased attesting the dysregulation of oxidative stress response. In conclusion this is the first quantitative study to highlight the involvement of endothelial dysfunction in CTEPH using patient samples and by network medicine approach

Bioanalytical and proteomic approaches in the study of pathologic ECs dysfunctionality, oxidative stress and the effect of PFKFB3 modulators

[eng] The vascular endothelium is one of the major targets of oxidative stress, playing an important role in the development of vascular-related disorders, such as atherosclerotic disease1. Oxidative stress increases the formation of reactive species (ROS), such as peroxides and hydroxyl radicals. ROS are signalling molecules and are able to induce leukocyte adhesion and promote the vascular endothelial permeability, that leads to the dysfunction of endothelium- related signal transduction pathways as well as redox-regulated transcription factors2. ROS cause reorganization of the actin filament, formation of an intracellular gap and changes in the cell shape. The endothelium contains adherents and tight junctions that are responsible for maintaining the restrictive barrier. Figure A1 shows the subcellular endothelial sites regulated by oxidative stress. In the normal structure of vascular endothelium, the lateral cell border contains occluding protein in association with zonula occludens-1 (ZO-1); VE-cadherin in association with a-, b-, and g-catenins; other proteins such as platelet endothelial cell adhesion molecule 1 (PECAM-1). Cortical actin filaments are directly linked with ZO-1, vinculin and α-actinin, that connect catenin to VE-cadherin in actin filament network. Intercellular adhesion molecule-1 (ICAM-1) expressed on the endothelial luminal surface might also involve in cortical actin filaments. ECs continuously express surface adhesion molecules including vascular cell adhesion molecule-1 (VCAM-1), ICAM-1, ICAM-2 and P-selectin (stored in Weible-Palade bodies). However, oxidative stress causes phosphorylation and reorganization of occludin and PECAM-1, reduces levels of vascular endothelium catenins and actin-binding proteins. In the actin filament network, oxidant stress causes the increased formation of stress fibers and decreased cortical actin band, resulting in disruption of the tight and adherens junctions. ROS promote cell contraction and stress fiber formation due to the increased phosphorylation of myosin light chain (MLC). Additionally, oxidative stress functionally upregulates adhesion molecule

Detection of viral RNA fragments in human iPSC cardiomyocytes following treatment with extracellular vesicles from SARS-CoV-2 coding sequence overexpressing lung epithelial cells

Coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a global pandemic. The prevalence/severity of COVID-19 is higher among patients with cardiovascular risk factors. Despite the expression of angiotensin-converting enzyme 2 (ACE2), a receptor for SARS-CoV-2 infection, in cardiomyocytes, there has been no conclusive evidence of direct viral infection although the presence of viral genome within COVID-19 patients' hearts has been reported. Here, we overexpressed SARS-CoV-2 genes in A549 lung epithelial cells. We then isolated extracellular vesicles (EVs) and detected the presence of viral RNA within these EVs. We observed that human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are receptive to these EVs, and viral genes were detectable in the cardiomyocytes. Accordingly, the uptake of viral RNA-harboring EVs led to an upregulation of inflammation-related genes in hiPSC-CMs. Thus, our findings indicate that SARS-CoV-2 RNA containing EVs represents an indirect route of viral RNA entry into cardiomyocytes.Open access articleThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]