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Retinal microglia in the mouse model of oxygen-induced retinopathy

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6 8 10 12 14 16 18 20 0

50 100 150 200 250 300 350

400 Hyalocytes

time (days after birth)

cells / mm²

6 8 10 12 14 16 18 20

0 100 200 300 400 500 600

Microglial cells in peripheral zone

time (days after birth)

cells / mm²

6 8 10 12 14 16 18 20

0 100 200 300 400 500 600

Microglial cells in central zone

time (days after birth)

cells / mm²

Retinal microglia in the mouse model of oxygen-induced retinopathy

Fischer F; Martin G; Agostini HT

University Eye Hospital Freiburg, Germany

Contact: G. Martin, gottfried.martin@uniklinik-freiburg.de

Retinal neovascularization has been

intensively investigated in the mouse model of oxygen-induced retinopathy (OIR). Here, we studied the contribution of microglial cells to cell loss and reactive angiogenesis in

mice expressing green fluorescent protein (GFP) under the microglial cell-specific

promotor Cx3cr1.

Background

Microglial cell density is always higher in the superficial than in the deep retinal layer.

Hyalocytes density decreases with retinal vascularisation.

The density of microglial cells in the deep layer decreases significantly after return to normal room air.

It is unclear if microglial cells move from the deep to the superficial layer.

Activated microglial cells are observed in the superficial retinal layer from P8 to P10 and from P16 to P18.

Thus, microglial cells may be involved in the formation of the central avascular zone as well as in retinal revascularisation.

Conclusion

Methods

Results

Animals: heterozygous Cx3cr1-GFP mice (background: C57Bl/6)

Retinal flat-mounts were stained with lectin and investigated by fluorescence microscopy

Numbers of microglial cells and hyalocytes in 12 fields were determined from postnatal day 7 to day 20

Cells were counted:

A) in two zones of the retina (central and peripheral zone) B) in two layers of the retina (superficial and deep layer) C) hyalocytes in vitreous

A) Retinal flat mount, Lectin staining, OIR P17

superficial layer

deep layer

activated

microglia cells

OIR-model

control group without oxygen treatment

central zone

peripheral zone

superficial layer

deep layer

Mouse model of oxygen-induced retinopathy (OIR)

superficial layer

deep layer

B) Cryosection of a OIR-treated mouse, OIR P17, GFP+ microglial cells (green)

room air

P0 P7 P12 P17

retinal vascularisation vessel loss neovascularisation regression of the NV

75 % O

2

room air

central avascular zone

peripheral zone

OIRcontrol group

Hyalocytes in the central avascular zone of the retina,

vessels stained with lectin

Hyalocytes are positive for GFP and lectin. In contrast to the microglial

cells, they have a larger round cellular body and no processes.

Activated microglial cells

OIR P17

These cells are found in the superficial layer of the retinal avascular zone.

Activated microglial cells express GFP and are additionally positive for lectin. They undergo a

morphological change from

ramified cells to cells with short and broad processes.

bar = 50μm Error bars indicate standard error

central zone P12 central zone P14

Microglial cells expressing GFP (green) in the deep retinal layer, vessels are stained with lectin (red)

bar = 50μm

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