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. 2003 Jul 9;23(14):6030–6040. doi: 10.1523/JNEUROSCI.23-14-06030.2003

Figure 1.

Figure 1.

Distribution of retinoschisin mRNA and protein in normal (A) and rd/rd mouse (B) retinas. A, Retinoschisin mRNA was detected with a digoxigenin-labeled antisense-cRNA probe (AS) and visualized by the purple reaction product of the alkaline phosphatase reaction; no staining was observed with sense-cRNA probe (S) used as control. Retinoschisin was detected by immunohistochemistry (IHC); the brown peroxidase-reaction product was observed after immunoreaction with an affinity-purified polyclonal antibody against amino acid residues 24–37 of retinoschisin (aa 24–37). This antibody was preabsorbed with aa 24–37 and used as control (PA). NBZ, Neuroblastic zone; IPL, inner plexiform layer; G/N, ganglion cell and nerve fiber layers; RPE, retinal pigment epithelium; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; C, choroid; OS, outer segment of photoreceptors; IS, inner segment of photoreceptors. Retinoschisin mRNA and protein were not detected in the retina of normal mice at P3 but were found at a very low level at P5. A mismatched mRNA and protein localization started on P7 and persisted into adulthood (P82). Retinoschisin mRNA was in the outer retina throughout postnatal development (outer NBZ on P5; ONL on P6 and P7/8; IS and ONL on P14, 21, and 82). In contrast, retinoschisin (Protein) was initially in the outer retina (outer NBZ on P5 and ONL on P6) and then appeared in the inner retina. Immunopositive cell bodies in the INL (double arrowheads) started appearing at P8 and showed increased intensity as the retina matured. By P21, retinoschisin was abundant in all layers. Both mRNA and protein were most concentrated in the area corresponding to the photoreceptor inner segments (*). Visualization of the mRNA was achieved with a 2 hr alkaline phosphatase reaction (P7/8, 14, and 21) or 10.5 hr (P3, 5, 6, and 82, and the left micrograph in the AS column for P14). B, At P73 and P139, most photoreceptors had degenerated in the rd/rd retinas, and retinoschisin mRNA was not detected even after 10.5 hr of reaction. Faint retinoschisin staining was present in the INL, the IPL, and the G/N. The micrograph strips at the far right of the mRNA and protein panels for each age are high-contrast bright-field micrographs taken from the same section as the adjacent micrographs. High-contrast micrographs were also taken from a portion of the sections in the AS panel at P3, the IHC panel at P3, and the AS panel at P5. Scale bar: for all micrographs, 50 μm.