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Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence
Published on: April 17, 2019
MCT-4, A511/Basigin and EF5 expression patterns during early chick cardiomyogenesis indicate cardiac cell
Mingda Han1, Patrizia Trotta, Carl Coleman
1Department of Pediatrics, USF-Children's Research Institute, St. Petersburg, Florida 33701, USA.
Insights
Hypoxia marker EF5 indicates chick heart cells differentiate anaerobically. Monocarboxylate transporters (MCTs) and Basigin support this, with MCT-4 and Basigin showing early expression patterns during heart development.
Area of Science:
- Developmental Biology
- Cellular Metabolism
- Cardiovascular Research
Background:
- Hypoxia is a known factor influencing cell differentiation.
- Monocarboxylate transporters (MCTs) facilitate lactate and pyruvate transport, crucial for energy metabolism.
- Basigin (5A11/CD147) is essential for MCTs' plasma membrane localization and function.
Purpose of the Study:
- To investigate the role of anaerobic metabolism in early chick heart development.
- To analyze the expression patterns of key glycolytic metabolism indicators during cardiac formation.
Main Methods:
- Immunohistochemical localization of hypoxia marker EF5.
- Analysis of MCT-4, MCT-1, and 5A11 (Basigin/CD147) expression during chick embryonic stages (4-12).
- RT-PCR to confirm gene expression throughout cardiac development.
Main Results:
- EF5, a hypoxia marker, is present in early chick heart fields (stage 4/5).
- MCT-4 and 5A11/Basigin are expressed early in the bilateral plate mesoderm with a left-right pattern during cardiac formation.
- MCT-4 and 5A11/Basigin show sustained expression in the looping heart, while MCT-1 is detected from embryonic day 3 onwards.
Conclusions:
- Developing chick heart precursor cells utilize anaerobic metabolism in a hypoxic environment.
- MCT-4 and 5A11/Basigin play significant roles in early cardiac development and energy production.
- The findings suggest a specialized metabolic adaptation for cardiac differentiation under hypoxic conditions.
Abstract:
We have identified the presence of the hypoxia marker EF5 in the stage 4/5 chick heart fields. This suggests that cardiac cell differentiation occurs in a relatively anaerobic environment. Monocarboxylate transporter (MCT) studies in adult cardiac myocytes have demonstrated that MCTs catalyze proton-linked pyruvate and lactate transport activity. 5A11/Basigin is an ancillary protein that targets MCTs to the plasma membrane for their function. MCT-4 expression is most evident in cells with a high glycolytic rate associated with hypoxic energy production. Subsequent to the immunohistochemical localization of EF5 in the early heart field, we continued in our analysis during stages 5 to 12 for the expression of indicators of cellular glycolytic metabolism in the developing heart, such as MCT-4, MCT-1, and 5A11 (Basigin/CD147). Our observations indicate that MCT-4 and 5A11/Basigin are expressed early, in a differential left-right pattern, in the bi-lateral plate mesoderm, as the cardiac compartment is forming. At stage 11, MCT-4/5A11 continues to be highly expressed in the myocardial wall of the looping heart, but not in the dorsal mesocardium. RT-PCR analyses for MCT-1, -4, and 5A11 indicate that MCT-4 and 5A11 are expressed throughout precardiac, embryonic, and fetal stages in the heart. MCT-1 is first detected in the heart on embryonic day 3 and then remains expressed throughout development to hatching. These results indicate that cardiac precursor cells are equipped for differentiating in a hypoxic environment using anaerobic metabolism for energy production.
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