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Published on: February 10, 2021
Complement expression in the retina is not influenced by short-term pressure elevation
Konstantin Astafurov1, Cecilia Q Dong2, Lampros Panagis1
1Department of Cell Biology, SUNY Downstate Medical Center, Brooklyn, NY.
Purpose:
To determine whether short-term pressure elevation affects complement gene expression in the retina in vitro and in vivo.
Methods:
Muller cell (TR-MUL5) cultures and organotypic retinal cultures from adult mice and monkeys were subjected to either 24-h or 72-h of pressure at 0, 15, 30, and 45 mmHg above ambient. C57BL/6 mice were subjected to microbead-induced intraocular pressure (IOP) elevation for 7 days. RNA and protein were extracted and used for analysis of expression levels of complement component genes and complement component 1, q subcomponent (C1q) and complement factor H (CFH) immunoblotting.
Results:
mRNA levels of complement genes and C1q protein levels in Muller cell cultures remained the same for all pressure levels after exposure for either 24 or 72 h. In primate and murine organotypic cultures, pressure elevation did not produce changes in complement gene expression or C1q and CFH protein levels at either the 24-h or 72-h time points. Pressure-related glial fibrillary acidic protein (GFAP) mRNA expression changes were detected in primate retinal organotypic cultures (analysis of variance [ANOVA]; p<0.05). mRNA expression of several other genes changed as a result of time in culture. Eyes subjected to microbead-induced IOP elevation had no differences in mRNA expression of complement genes and C1q protein levels (ANOVA; p>0.05 for both) with contralateral control and naïve control eyes.
Conclusions:
Short-term elevation of pressure in vitro as well as short-term (1 week) IOP elevation in vivo does not seem to dramatically alter complement system gene expression in the retina. Prolonged expression to elevated pressure may be necessary to affect the complement system expression.
Insights
Short-term pressure elevation in the retina does not significantly alter complement gene expression. Longer exposure to elevated pressure may be required to impact the complement system.
Area of Science:
- Ophthalmology
- Immunology
- Molecular Biology
Background:
- The complement system plays a crucial role in ocular immune responses.
- Understanding how intraocular pressure (IOP) affects retinal complement gene expression is vital for diagnosing and treating glaucoma and other pressure-related eye diseases.
Purpose of the Study:
- To investigate the impact of short-term elevated pressure on retinal complement gene expression in vitro and in vivo.
- To determine if acute pressure changes influence key complement components like C1q and CFH.
Main Methods:
- Cultured Muller cells and organotypic retinal cultures (murine and primate) were exposed to varying pressure levels (0-45 mmHg) for 24 or 72 hours.
- Mice underwent microbead-induced IOP elevation for 7 days.
- Gene and protein expression of complement components, C1q, and CFH were analyzed using RNA and immunoblotting.
Main Results:
- No significant changes in complement gene mRNA or C1q protein levels were observed in Muller cell cultures under elevated pressure.
- Organotypic retinal cultures from mice and primates showed no alterations in complement gene expression or C1q/CFH protein levels after short-term pressure exposure.
- Microbead-induced IOP elevation in mice did not lead to significant changes in retinal complement gene expression or C1q protein levels.
Conclusions:
- Short-term elevation of intraocular pressure, both in vitro and in vivo, does not appear to substantially alter retinal complement gene expression.
- Prolonged exposure to elevated pressure may be necessary to elicit a significant response in the retinal complement system.
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