Fig. 4. Hypoxia in the retina after RD.
(A) Representative immunohistochemistry images of the outer nuclear layer labeled with Hypoxyprobe (brown staining) comparing the detached portion of a mouse retina (right panel) to the attached portion of the same retina (left panel) 24 hours after RD. Both sides of the retina were stained with toluidine blue stain (n = 7). (B) In vivo oxygen concentrations taken in the retina 24 hours after RD in an attached retina (right eye) compared to the detached retina (left eye) (n = 7). (C) Representative TUNEL labeling in the outer nuclear layer 24 hours after RD in mice kept in room air (left panel) compared to mice kept in 75% oxygen (right panel). (D) Quantitation of TUNEL-positive cells in mice kept in room air (n = 5) for 24 hours after RD compared to mice kept in 75% oxygen (n = 6). (E) RT-PCR showing gene expression for Fb (encoding factor B) in the attached or detached retina 24 hours after RD; comparing mice kept in room air to mice kept in 75% oxygen (attached, n = 3; room air, n = 3; 75% O2, n = 4). (F) Under hypoxic conditions (1% O2), increasing mouse serum concentrations (used as a source of complement) led to an increased percentage of cell death in 661W cells assessed using a live/dead assay (n = 4). *P ≤ 0.05, ****P ≤ 0.0001; unpaired Student’s t test. Scale bars, 50 μm.