Related Experiment Video
Updated: May 15, 2026
Primary Cell Cultures to Study the Regeneration Potential of Murine Müller Glia after MicroRNA Treatment
Published on: March 28, 2022
Apelin-13 regulates proliferation, migration and survival of retinal Müller cells under hypoxia
Qiang Lu1, Yan-Rong Jiang, Jing Qian
1Department of Ophthalmology, People's Hospital, Peking University, Beijing, China.
Aims:
To investigate the effect of apelin-13 and the antagonist of apelin receptor (F13A) on retinal Müller cells in vitro.
Methods:
Localization of apelin-13, GFAP and VEGF of Müller cells was detected by immunofluorescence. The effects of apelin-13 and F13A on cell function were assessed by MTT, spreading assay, apoptosis and Boyden chamber assay in vitro. Additionally, the mRNA and protein of apelin-13, GFAP and VEGF in cultured Müller cells were measured by real-time PCR and western blot.
Results:
Under hypoxia, strong positive staining of apelin-13 was observed and particularly evident in the cytosol and around the nucleus. Exposure of Müller cells to hypoxia led to a progressive increase in mRNA (p<0.01) and protein levels of apelin-13 (p<0.01), with a maximal 2.5-fold and 2-fold stimulation at 4h respectively, compared with normoxic controls. Treated with 0.1, 1, 10 and 100 ng/ml apelin-13, the protein level of GFAP (p<0.01) and VEGF (p<0.01) increased significantly in Müller cells in a dose-dependent manner after 24h. Compared with the untreated cells, 10 ng/ml apelin-13 significantly promoted Müller cells migration (p<0.01). Annexin/PI staining showed that apelin-13 can downregulate cell apoptosis with 30% to the most (p<0.05). On the contrary, 20 ng/ml F13A-treated Müller cells spread less than the control cells, with significantly lower number of migrated cells and significantly higher rate of apoptosis.
Conclusions:
The results of this study showed that apelin-13 modulated the proliferation, migration, spreading, survival of Müller cells and the expressions of GFAP and VEGF.
Insights
Apelin-13 promotes retinal Müller cell proliferation, migration, and survival while upregulating GFAP and VEGF. Its antagonist, F13A, has opposite effects, indicating apelin-13’s role in Müller cell function.
Area of Science:
- Ophthalmology
- Cell Biology
- Molecular Biology
Background:
- Retinal Müller cells play crucial roles in retinal homeostasis and disease.
- Apelin-13 is a peptide hormone with diverse physiological functions.
- The role of apelin-13 in retinal Müller cells remains largely unexplored.
Purpose of the Study:
- To investigate the in vitro effects of apelin-13 and its receptor antagonist (F13A) on retinal Müller cells.
- To determine the impact of apelin-13 on Müller cell proliferation, migration, spreading, and apoptosis.
- To examine the influence of apelin-13 on the expression of glial fibrillary acidic protein (GFAP) and vascular endothelial growth factor (VEGF) in Müller cells.
Main Methods:
- Immunofluorescence was used to detect apelin-13, GFAP, and VEGF.
- Cell proliferation, spreading, apoptosis, and migration were assessed using MTT, spreading assays, Annexin/PI staining, and Boyden chamber assays.
- mRNA and protein levels of apelin-13, GFAP, and VEGF were quantified by real-time PCR and Western blot.
Main Results:
- Hypoxia increased apelin-13 mRNA and protein levels in Müller cells.
- Apelin-13 dose-dependently increased GFAP and VEGF protein levels and promoted cell migration and spreading.
- Apelin-13 reduced Müller cell apoptosis, while F13A exhibited opposing effects.
Conclusions:
- Apelin-13 significantly modulates Müller cell proliferation, migration, spreading, and survival.
- Apelin-13 influences the expression of GFAP and VEGF in retinal Müller cells.
- These findings highlight apelin-13 as a potential therapeutic target in retinal diseases involving Müller cells.
