(a) Expression of Clcc1 mRNA in the cornea, lens, iris, optic nerve, and retina by qRT-PCR at different ages. Values represent the mean (± SD) on an arbitrary scale (y axis) and were calculated from at least three independent experiments. While Clcc1 is expressed in all tissues tested, ocular expression is greatest in the retina and least in the lens. (b-f) In situ hybridization of clcc1 probes in zebrafish. clcc1 Is expressed widely in zebrafish. Staining with a digoxigenin-labeled cRNA probe shows a strong signal (black arrows) in the hindbrain (HB), swim bladder (SB), and eye at 1 dpf (b, c), and in the ganglion cell layer (GCL), outer nuclear layer (ONL), and retinal pigmented epithelium (RPE) at 3 dpf (d, e and f); OS. Scale Bar: 100 μm. (g–j) IHC of formalin fixed and paraffin embedded human retinal sections demonstrated CLCC1 is expressed extensively in the retina and optical nerves. High magnification (g,i) shows more intense CLCC1 staining (arrow) in the lamina cribrosa (LC), optic nerve (ON), ganglion cell layer (GCL), inner nuclear layer (INL), outer nuclear layer (ONL) and retinal pigmented epithelia (RPE) in the retina (counter stain is methyl green). Scale Bar: g, h, 50 μm; i, j, 20μm.