The spatiotemporal overview of retinoschisin (Rs1) protein presence during the first five days of retinal development shows that Rs1 protein presence is first detected at 48 hpf and increases in intensity and spatial distribution up to 120 hpf. Cryosections were incubated with primary antibodies overnight at 4°C. At least 3 biological replicates were examined per timepoint. A: No positive immunostaining of Rs1 was observed within the first 24 h of retinal development. B: At 48 h, Rs1 protein presence was first observed in this experiment in low levels in the ganglion cell layer (GCL). C: At 60 hpf, a weak signal was detected in the GCL, the inner plexiform layer (IPL), and the outer nuclear layer (ONL) D‐E. The intensity of Rs1 immunostaining continued to increase throughout 72 (D), 96 (E), and 120 (F) hpf. G. Co‐staining of Rs1 and cone arrestin (Arr3) demonstrated that Rs1 surrounds the developing outer segments at 120 hpf. F. Rs1 presence was quantified per retinal layer and normalized to total Rs1 signal per sample. Data represent mean percentage of Rs1 signal per layer (± standard error of the mean). A shift in Rs1 localization is observed from predominately INL at 72 hpf toward increasing IPL and GCL contribution at later stages. Hpf = hours post fertilization, Rs1 = retinoschisin, Arr3 = cone arrestin, DAPI = 4′,6‐diamidino‐2‐phenylindole.
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