Figure 2: Adamts9 mRNA is expressed during murine eye development consistent with a developmental sequence of ASD in Adamts9^del/+ eyes.

(a) In situ hybridization using an Adamts9 probe was performed on E10.5 through newborn (P0) Adamts9^+/+ eyes. Adamts9 mRNA expression is visualized as red dots overlying cells, against a purple hematoxylin counterstain. Adamts9 was expressed throughout this period in the anterior pole of the retina (arrows) and hyaloid vasculature (single asterisk). Adamts9 expression was also observed in the lens and the lens epithelium (arrowheads) and in mesenchymal cells within the developing cornea (double asterisk). L = lens, R = optic cup/retina, I = iris, C = cornea. The boxed areas in the upper panels are shown at higher magnification (lower black boxed panels). Scale bar = 50 μm; blue scale bar in P0 images = 250 μm. (b,c) Sections from Adamts9^+/+ and Adamts9^del/+ eyes from E14.5 to P0 were stained by H&E and the lens area was measured. At E14.5 Adamts9^+/+ and Adamts9^del/+ eyes were of comparable size. Posterior to the lens, abnormal material (indicated by an arrow) was observed in the E16.5 and P0 Adamts9^del/+ eyes shown here (b) but never in Adamts9^+/+ eyes. At E16.5 and P0, the Adamts9^del/+ lens was significantly smaller than the Adamts9^+/+ lens (c). Images are representative of at least 3 eyes analyzed at each time point. L = lens. Scale bar = 100 μm. Significance was determined using a 2-tailed Student’s t test (*p < 0.05).