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Updated: Apr 28, 2026
Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo
Published on: July 9, 2013
Neonatal systemic inflammation in rats alters retinal vessel development and simulates pathologic features of
Hye Kyoung Hong, Hyun Ju Lee, Jung Hwa Ko
1Department of Ophthalmology, Seoul National University Bundang Hospital, 300 Gumi-dong, Seongnam 463-707, Korea. sejoon1@snu.ac.kr.
Insights
Systemic inflammation in neonatal rats, induced by lipopolysaccharide (LPS), impaired retinal vessel development and caused inflammation, mimicking retinopathy of prematurity (ROP). This suggests perinatal inflammation may contribute to ROP pathogenesis.
Area of Science:
- Ophthalmology
- Neonatal development
- Inflammation research
Background:
- Retinopathy of prematurity (ROP) is a leading cause of childhood visual impairment.
- ROP is linked to altered retinal angiogenesis in preterm infants.
- Perinatal infections and inflammation are associated with increased ROP rates.
Purpose of the Study:
- To investigate the effects of systemic inflammation on retinal vessel development.
- To examine retinal inflammation in neonatal rats exposed to systemic inflammation.
Main Methods:
- Neonatal rats received intraperitoneal injections of lipopolysaccharide (LPS) or saline on postnatal days 1, 3, and 5.
- Retinas were analyzed on postnatal days 7 and 14 for vascular changes, inflammatory cells, molecules, and apoptosis.
Main Results:
- LPS injection impaired retinal angiogenesis, decreasing vessel extension and capillary density.
- Abnormal vessel growth, dilated peripheral vascular ridge, and inflammatory cell infiltration were observed.
- Increased levels of inflammatory markers (TNF-α, IL-1β, IL-12a) and apoptosis were found in LPS-treated retinas.
Conclusions:
- Systemic LPS-induced inflammation causes retinal inflammation and angiogenesis impairment in neonatal rats.
- Perinatal inflammation is implicated as a contributing factor in ROP pathogenesis.
Background:
Alteration of retinal angiogenesis during development leads to retinopathy of prematurity (ROP) in preterm infants, which is a leading cause of visual impairment in children. A number of clinical studies have reported higher rates of ROP in infants who had perinatal infections or inflammation, suggesting that exposure of the developing retina to inflammation may disturb retinal vessel development. Thus, we investigated the effects of systemic inflammation on retinal vessel development and retinal inflammation in neonatal rats.
Methods:
To induce systemic inflammation, we intraperitoneally injected 100 μl lipopolysaccharide (LPS, 0.25 mg/ml) or the same volume of normal saline in rat pups on postnatal days 1, 3, and 5. The retinas were extracted on postnatal days 7 and 14, and subjected to assays for retinal vessels, inflammatory cells and molecules, and apoptosis.
Results:
We found that intraperitoneal injection of LPS impaired retinal vessel development by decreasing vessel extension, reducing capillary density, and inducing localized overgrowth of abnormal retinal vessels and dilated peripheral vascular ridge, all of which are characteristic findings of ROP. Also, a large number of CD11c+ inflammatory cells and astrocytes were localized in the lesion of abnormal vessels. Further analysis revealed that the number of major histocompatibility complex (MHC) class IIloCD68loCD11bloCD11chi cells in the retina was higher in LPS-treated rats compared to controls. Similarly, the levels of TNF-α, IL-1β, and IL-12a were increased in LPS-treated retina. Also, apoptosis was increased in the inner retinal layer where retinal vessels are located.
Conclusions:
Our data demonstrate that systemic LPS-induced inflammation elicits retinal inflammation and impairs retinal angiogenesis in neonatal rats, implicating perinatal inflammation in the pathogenesis of ROP.
