Figure 5.

Late loss of bipolar cells was counteracted by hypothermia. (A) CALB mRNA expression was measured via qPCR. After eight days, a significantly decreased CALB mRNA expression was seen in the hypothermia treated CoCl₂ + 30 °C retinae. (B) Representative images of bipolar cells stained with anti-Chx10 (green) at eight days. DAPI was used for the visualization of cell nuclei (blue). (C) After eight days, all groups had a similar number of Chx10⁺ cells. (D) Representative pictures of the inner layers are given. Rod bipolar cells (red) were stained immunohistochemically at days four and eight using an anti-PKCα antibody. Cell nuclei are shown in blue. (E) At day eight, a significant loss of bipolar cells was noted in the CoCl₂ + 37 °C. A rescue of PKCα⁺ cells was achieved by hypothermia. Abbreviations: GCL = ganglion cell layer; IPL = inner plexiform layer; INL = inner nuclear layer; OPL = outer plexiform layer; ONL = outer nuclear layer; qPCR = quantitative real-time PCR; IHC = immunohistochemistry. Values are mean ± SEM. A: n = 6–7/group; C,E: n = 9–10/group. Statistical differences to control + 37 °C group are marked with * and differences to CoCl₂ + 37 °C group with ^#. ^#,*p < 0.05; **p < 0.01 Scale bar = 20 µm.