epon-post-embedding-correlative-light-and-electron-microscopy

Epon Post Embedding Correlative Light and Electron Microscopy

Correlative light and electron microscopy (CLEM) is a comprehensive microscopy that combines the localization information provided by fluorescence ...

in-vivo-imaging-of-zebrafish-larvae-heart-regeneration

<em>In Vivo</em> Imaging of Zebrafish Larvae Heart Regeneration

Cardiovascular disease is the leading cause of death globally, so learning about regeneration in the heart is a critical research topic. In the past, ...

Fujian Medical University

Fujian Maternity and Child Health Hospital

Bepridil is a commonly used medication for arrhythmia and heart failure. It primarily exerts hemodynamic effects by inhibiting Na/K movement and regulating the Na/Ca exchange. In comparison to other Ca inhibitors, bepridil has a long half-life and a complex pharmacology. Additionally, it is widely used in antiviral research and the treatment of various diseases. However, the toxicity of this compound and its other possible effects on embryonic development are unknown. In this study, we investigated the toxicity of bepridil on rat myocardial H9c2 cells. After treatment with bepridil, the cells became overloaded with Ca and entered a state of cytoplasmic vacuolization and nuclear abnormality. Bepridil treatment resulted in several morphological abnormalities in zebrafish embryo models, including pericardium enlargement, yolk sac swelling, and growth stunting. The hemodynamic effects on fetal development resulted in abnormal cardiovascular circulation and myocardial weakness. After inhibiting the Ca transmembrane, the liver of zebrafish larvae also displayed an ectopic and deficient spatial location. Additionally, the results of the RNA-seq analysis revealed the detailed gene expression profiles and metabolic responses to bepridil treatment in zebrafish embryonic development. Taken together, our study provides an important evaluation of antiarrhythmic agents for clinical use in prenatal heart patients.

Effects of bepridil on early cardiac development of zebrafish.

Congenital heart disease (CHD) is the most common type of birth defect and the main noninfectious cause of death during the neonatal stage. The non-POU domain containing, octamer-binding gene, NONO, performs a variety of roles involved in DNA repair, RNA synthesis, transcriptional and post-transcriptional regulation. Currently, hemizygous loss-of-function mutation of NONO have been described as the genetic origin of CHD. However, essential effects of NONO during cardiac development have not been fully elucidated. In this study, we aim to understand role of Nono in cardiomyocytes during development by utilizing the CRISPR/Cas9 gene editing system to deplete Nono in the rat cardiomyocytes H9c2. Functional comparison of H9c2 control and knockout cells showed that Nono deficiency suppressed cell proliferation and adhesion. Furthermore, Nono depletion significantly affected the mitochondrial oxidative phosphorylation (OXPHOS) and glycolysis, resulting in H9c2 overall metabolic deficits. Mechanistically we demonstrated that the Nono knockout impeded the cardiomyocyte function by attenuating phosphatidyl inositol 3 kinase-serine/threonine kinase (Pi3k/Akt) signaling via the assay for transposase-accessible chromatin using sequencing in combination with RNA sequencing. From these results we propose a novel molecular mechanism of Nono to influence cardiomyocytes differentiation and proliferation during the development of embryonic heart. We conclude that NONO may represent an emerging possible biomarkers and targets for the diagnosis and treatment of human cardiac development defects.

Nono deficiency impedes the proliferation and adhesion of H9c2 cardiomyocytes through Pi3k/Akt signaling pathway.

Fluoride has strong electronegativity and exposes diversely in nature. Water fluoridation is the most pervasive form of occurrence, representing a significant threat to human health. In this study, we investigate the morphometric and physiological alterations triggered by fluoride stimulation during the embryogenesis of zebrafish and reveal its putative effects of stage- and/or dose-dependent. Fluoride exhibits potent biological activity and can be extensively absorbed by the yolk sac, exerting significant effects on the development of multiple organs. This is primarily manifested as restricted nutrient utilization and elevated levels of lipid peroxidation, further leading to the accumulation of superoxide in the yolk sac, liver, and intestines. Moreover, pericardial edema exerts pressure on the brain and eye development, resulting in spinal curvature and reduced body length. Besides, acute fluoride exposure with varying concentrations has led to diverse teratogenic outcomes. A low dose of water fluoridation tends to induce abnormal development of the embryonic yolk sac, while vascular malformation is widely observed in all fluoride-treated groups. The effect of fluoride exposure on blood circulation is universally present, even in zebrafish larvae that do not exhibit obvious deformities. Their swimming behavior is also affected by water fluoridation, resulting in reduced activity and delayed reactions. In conclusion, this study provides valuable insights into the monitoring of environmental quality related to water fluoridation and disease prevention.

Effects of water fluoridation on early embryonic development of zebrafish.

 var.  (RW) is one of the traditional Chinese medicinal materials, which is used to treat angina pectoris (AP). However, the possible underlying mechanisms remains unclear. The aim of this study was to explore RW in the treatment of AP and to identify the potential mechanism of the core compounds.

Investigating the multitarget pharmacological mechanism of  var.  acting on angina pectoris using combined network pharmacology and molecular docking.

JOURNAL/nrgr/04.03/01300535-202501000-00034/figure1/v/2024-05-14T021156Z/r/image-tiff Certain amino acids changes in the human Na+/K+-ATPase pump, ATPase Na+/K+ transporting subunit alpha 1 (ATP1A1), cause Charcot-Marie-Tooth disease type 2 (CMT2) disease and refractory seizures. To develop in vivo models to study the role of Na+/K+-ATPase in these diseases, we modified the Drosophila gene homolog, Atpα, to mimic the human ATP1A1 gene mutations that cause CMT2. Mutations located within the helical linker region of human ATP1A1 (I592T, A597T, P600T, and D601F) were simultaneously introduced into endogenous DrosophilaAtpα by CRISPR/Cas9-mediated genome editing, generating the AtpαTTTF model. In addition, the same strategy was used to generate the corresponding single point mutations in flies (AtpαI571T, AtpαA576T, AtpαP579T, and AtpαD580F). Moreover, a deletion mutation (Atpαmut) that causes premature termination of translation was generated as a positive control. Of these alleles, we found two that could be maintained as homozygotes (AtpαI571T and AtpαP579T). Three alleles (AtpαA576T, AtpαP579 and AtpαD580F) can form heterozygotes with the Atpαmut allele. We found that the Atpα allele carrying these CMT2-associated mutations showed differential phenotypes in Drosophila. Flies heterozygous for AtpαTTTF mutations have motor performance defects, a reduced lifespan, seizures, and an abnormal neuronal morphology. These Drosophila models will provide a new platform for studying the function and regulation of the sodium-potassium pump.

Drosophila models used to simulate human ATP1A1 gene mutations that cause Charcot-Marie-Tooth type 2 disease and refractory seizures.

The C.florida. is one of the common medicines used by She population in China, with therapeutic effects of promoting blood circulation and anti-inflammatory. According to the acute toxicity grading standard of chemical substances, this herb is a low-toxicity herb. At present, the safety of C.florida., especially its impact on early embryonic development, is still unclear.

Early developmental anomalies in zebrafish (Danio rerio) embryos induced by the Clematis florida Thunb.

Valproic acid (VPA) is predominantly prescribed for epilepsy, convulsions, and other psychiatric disorders. As an epigenetic regulator, it is also used to treat various forms of cancer. The clinical demand for the drug may pose an environmental hazard. Evidence indicates that VPA's significant therapeutic value comes at the cost of possible side toxic effects, as symptoms of birth defects have been confirmed in animal experiments using VPA. However, the effects of VPA during the development of the circulatory system remain unclear. In this study, zebrafish embryos were exposed to a series of concentrations of VPA between three hours post fertilization (hpf) and five days post fertilization (dpf). The results demonstrated time- and dose-dependent developmental delays in the zebrafish, including cardiovascular malformation and decreased movement and reaction time. Consistent with the in vivo results, exposure to VPA increased the levels of myocardial reactive oxygen species (ROS) and cell apoptosis through cardiac mitochondrial turnover disorders. The expression levels of genes related to cardiovascular development and antioxidant response were downregulated, while genes related to apoptosis pathways were upregulated. Overall, our toxicological studies of VPA exposure illustrate the damage to cardiovascular development, raising concerns about the hazard of VPA exposure in early pregnancy. Our study provides novel insights into the potential environmental risks of VPA.

Cardiovascular developmental hazards of valproic acid in zebrafish.

Polycystic ovary syndrome (PCOS) and premature ovarian failure (POF) are common causes of female infertility, both characterized by endocrine and metabolic dysregulation and anovulation. The She ethnomedicine formula, Medical Research Center Sheyao No. 1 (MRCSY1#), traditionally used for reproductive disorders, remains mechanistically underexplored.

ArticleTotal : 2	Year
Publication title Cited by 1	2024
Publication title	2026

Article	Year
Effects of bepridil on early cardiac development of zebrafish. Cell and tissue research\| PubMed ID: 36422735	2023
Nono deficiency impedes the proliferation and adhesion of H9c2 cardiomyocytes through Pi3k/Akt signaling pathway. Scientific reports\| PubMed ID: 37130848	2023
Effects of water fluoridation on early embryonic development of zebrafish. Ecotoxicology and environmental safety\| PubMed ID: 38176185	2024
Investigating the multitarget pharmacological mechanism of var. acting on angina pectoris using combined network pharmacology and molecular docking. Journal of thoracic disease\| PubMed ID: 38505080	2024
Drosophila models used to simulate human ATP1A1 gene mutations that cause Charcot-Marie-Tooth type 2 disease and refractory seizures. Neural regeneration research\| PubMed ID: 38767491	2025
Early developmental anomalies in zebrafish (Danio rerio) embryos induced by the Clematis florida Thunb. Journal of ethnopharmacology\| PubMed ID: 39369922	2025
Cardiovascular developmental hazards of valproic acid in zebrafish. Ecotoxicology and environmental safety\| PubMed ID: 39405976	2024
She ethnomedicine ameliorates ovarian dysfunction through enhanced cellular proliferation and anti-inflammation. Journal of ethnopharmacology\| PubMed ID: 40669675	2025

Xudong Zhuang

Biography

JoVE Journal Publications

Other Publications