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Quantification of Immunostained Caspase-9 in Retinal Tissue
Published on: July 25, 2022
Caspase-14: a novel caspase in the retina with a potential role in diabetic retinopathy
Mohamed Al-Shabrawey1, Saif Ahmad, Sylvia Megyerdi
1Department of Oral Biology and Anatomy, Georgia Health Sciences University (GHSU), College of Dental Medicine, Augusta, GA 30912, USA. malshabrawey@georgiahealth.edu
Purpose:
The purpose of this study was to evaluate caspase-14 expression in the retina under normal and diabetic conditions, and to determine whether caspase-14 contributes to retinal microvascular cell death under high glucose conditions.
Methods:
Quantitative real-time polymerase chain reaction and western blot analysis were used to evaluate caspase-14 expression in retinal cells, including pericytes (PCs), endothelial cells (ECs), astrocytes (ACs), choroidal ECs, and retinal pigment epithelium (RPE) cells. We also determined caspase-14 expression in the retinas of human subjects with or without diabetic retinopathy (DR) and in experimental diabetic mice. Retinal ECs and PCs were infected with adenoviruses expressing human caspase-14 or green fluorescent protein. Caspase-14 expression was also assessed in retinal vascular cells cultured under high glucose conditions. The number of apoptotic cells was determined with terminal deoxynucleotidyl transferase dUTP nick end labeling staining and confirmed by determining the levels of cleaved poly (ADP-ribose) polymerase-1 and caspase-3.
Results:
Our experiments demonstrated that retinal ECs, PCs, ACs, choroidal ECs, and RPE cells expressed caspase-14, and DR was associated with upregulation and/or activation of caspase-14 particularly in retinal vasculature. High glucose induced marked elevation of the caspase-14 level in retinal vascular cells. There was a significant increase in the apoptosis rate and the levels of cleaved poly (ADP-ribose) polymerase-1 and caspase-3 in retinal ECs and PCs overexpressing caspase-14.
Conclusions:
Our findings indicate that caspase-14 might play a significant role in the pathogenesis of DR by accelerating retinal PC and EC death. Further investigations are required to elaborate the underlying mechanisms.
Insights
Caspase-14 is upregulated in diabetic retinopathy (DR) and contributes to retinal microvascular cell death under high glucose conditions, suggesting a role in DR pathogenesis.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Diabetic retinopathy (DR) is a leading cause of vision loss.
- The role of specific cellular pathways, such as caspase-14, in DR pathogenesis remains incompletely understood.
Purpose of the Study:
- To evaluate caspase-14 expression in retinal cells under normal and diabetic conditions.
- To determine if caspase-14 contributes to retinal microvascular cell death induced by high glucose.
Main Methods:
- Quantitative real-time PCR and Western blot analysis were used to assess caspase-14 expression in various retinal cell types (pericytes, endothelial cells, astrocytes, RPE cells).
- Caspase-14 expression was examined in human retinas with and without DR, experimental diabetic mice, and cultured retinal vascular cells under high glucose.
- Apoptosis was quantified using TUNEL staining and assessment of cleaved PARP-1 and caspase-3 levels.
Main Results:
- Caspase-14 was expressed in multiple retinal cell types and was upregulated/activated in the retinal vasculature of DR patients and diabetic mice.
- High glucose significantly elevated caspase-14 levels in retinal vascular cells.
- Overexpression of caspase-14 in retinal endothelial cells and pericytes led to increased apoptosis.
Conclusions:
- Caspase-14 plays a significant role in DR pathogenesis by promoting retinal pericyte and endothelial cell death.
- Further research is needed to elucidate the precise mechanisms by which caspase-14 contributes to DR.
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01:27Diabetic Retinopathy
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