Cell adhesion to the extracellular matrix and its natural outcome of cell spreading, along with the maintenance of barrier activity, are essential behaviors of epithelial cells, including retinal pigment epithelium (RPE). Disruptions in these characteristics can result in severe vision-threatening diseases such as diabetic macular edema and age-related macular degeneration. However, the precise mechanisms underlying how RPE cells regulate their barrier integrity and cell spreading are not fully understood. This study aims to elucidate the relative importance of upper glycolytic components in governing these cellular behaviors of RPE cells. Electric Cell-Substrate Impedance Sensing (ECIS) technology was utilized to assess in real-time the effects of targeting various upper glycolytic enzymes on RPE barrier function and cell spreading by measuring cell resistance and capacitance, respectively. Specific inhibitors used included WZB117 for Glut1 inhibition, Lonidamine for Hexokinase inhibition, PFK158 for PFKFB3/PFK axis inhibition, and TDZD-8 for Aldolase inhibition. Additionally, the viability of RPE cells was evaluated using a lactate dehydrogenase (LDH) cytotoxicity assay. The most significant decrease in electrical resistance and increase in capacitance of RPE cells were observed due to dose-dependent inhibition of Glut1 using WZB117, as well as Aldolase inhibition with TDZD-8. LDH level analysis at 24-72 h post-treatment with WZB117 (1 and 10 μM) or TDZD-8 (1 μM) showed no significant difference compared to the control, indicating that the disruption of RPE functionality was not attributed to cell death. Lastly, inhibition of other upper glycolytic components, including PFKFB3/PFK with PFK158 or Hexokinase with Lonidamine, did not significantly affect RPE cell behavior. This study provides insights into the varied roles of upper glycolytic components in regulating the functionality of RPE cells. Specifically, it highlights the critical roles of Glut1 and Aldolase in preserving barrier integrity and promoting RPE cell adhesion and spreading. Such understanding will guide the development of safe interventions to treat RPE cell dysfunction in various retinal disorders.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11324787 | PMC |
| http://dx.doi.org/10.1038/s41598-024-68343-5 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Role of Fatty Acids β-Oxidation in the Metabolic Interactions Between Organs.
Int J Mol Sci
November 2024
Vanderbilt University Medical Center, Nashville, TN 37232, USA.
In recent decades, several discoveries have been made that force us to reconsider old ideas about mitochondria and energy metabolism in the light of these discoveries. In this review, we discuss metabolic interaction between various organs, the metabolic significance of the primary substrates and their metabolic pathways, namely aerobic glycolysis, lactate shuttling, and fatty acids β-oxidation. We rely on the new ideas about the supramolecular structure of the mitochondrial respiratory chain (respirasome), the necessity of supporting substrates for fatty acids β-oxidation, and the reverse electron transfer via succinate dehydrogenase during β-oxidation.
View Article and Find Full Text PDFGlucose-independent human cytomegalovirus replication is supported by metabolites that feed upper glycolytic branches.
Proc Natl Acad Sci U S A
November 2024
BIO5 Institute, University of Arizona, Tucson, AZ 85719.
Viruses with broad tissue distribution and cell tropism successfully replicate in various nutrient environments in the body. Several viruses reprogram metabolism for viral replication. However, many studies focus on metabolic reprogramming in nutrient-rich conditions that do not recapitulate physiological environments in the body.
View Article and Find Full Text PDFFatty acids promote migration of CD4 T cells through calcium release-activated calcium modulator ORAI1 sensitive glycolysis in dairy cows.
J Dairy Sci
October 2024
College of Veterinary Medicine, China Agricultural University, Beijing 100000. Electronic address:
Nutritional and metabolic state in dairy cows are important determinants of the immune response. During the periparturient period, a state of negative energy balance in the cow increases plasma concentrations of fatty acids, which are associated with inflammation. Among immune cells, CD4 T are able to function under high fatty acid (High-FA) conditions, but the underlying mechanisms regulating these events remain unclear.
View Article and Find Full Text PDFArticle Synopsis
- DOCK8 deficiency is a primary immunodeficiency caused by mutations in exon 45 of the DOCK8 gene, leading to symptoms like high serum IgE, eczema, and increased infections.
- Using CRISPR/Cas9, researchers created a mouse model with the same exon 45 mutation to study its effects on T cell behavior.
- The study found that the mutation affects T cell homeostasis, disrupts regulatory T cell development, alters glycolysis in CD4 T cells, and promotes a Th2 response upon infection, providing insights for potential treatments.
The tumor suppressor Parkin exerts anticancer effects through regulating mitochondrial GAPDH activity.
Oncogene
October 2024
Cancer Center, Department of Medical Oncology, Key Laboratory of Tumor Molecular Diagnosis and Individualized Medicine of Zhejiang Province, Key Laboratory for Diagnosis and Treatment of Upper Limb Edema and Stasis of Breast Cancer, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Hangzhou, China.
Cancer cells preferentially utilize glycolysis for energy production, and GAPDH is a critical enzyme in glycolysis. Parkin is a tumor suppressor and a key protein involved in mitophagy regulation. However, the tumor suppression mechanism of Parkin has still not been elucidated.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!