XB-IMG-154082
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Fig. 2.
MD3 KD disrupts eye morphogenesis. Alteration of eye morphology was analysed (A-E′) and quantified (F) in non-injected (NI), CT, MD3A, MD3B, MD3AB MO-injected embryos and embryos co-injected with MD3A MO and FL MD3 mRNA or MD3B MO and 7-mut MD3 mRNA. Lateral (A-E) and dorsal view (A′-E′) of the embryo; white arrow, level for dorsal view; black rectangle, eye; red asterisk, injected side of the embryo in all figures. Numbers indicate the embryos analysed. Crop of the eye (G-H) and head transversal section without (G′-H′) or with a hematoxylin & eosin (H&E) staining (G′′-H′′) were analysed in wild-type (wt) and MD3B MO embryos. Red arrow, retinal pigment epithelium (RPE); white arrowhead, eye region; dotted red line, remaining eye tissue. Analysis by immunohistochemistry on eye sections from wt and MD3B MO-injected embryos was performed for lens cells (β-crystallin, I-I′), rod photoreceptors (Rhodopsin; J,J′) and ganglion cells (Hu-C/Hu-D, K-K′), and the number of positive cells for each eye marker was quantified as described in the Materials and methods (L); numbers indicate the eye sections quantified. Statistical significance was assessed by Student's t-test in F (P values, CT MO=0.14; MD3A MO=8.99*10−5; MD3B MO=2.21*10−5; MD3AB MO=2.74*10−6; MD3A MO+MD3 FL RNA=0.18; MD3B MO+7mut-MD3 RNA=6*10−2, NS: non significant; ***P<0.001), and Mann–Whitney test in L (P values, β-crystallin=1.59*10−2; Rhodopsin=1*10−4; Hu-C/Hu-D=1.92*10−2, *P<0.05 and **P<0.01). Scale bars: 500 µm (A-E,G,H) and 100 μm (I-K). Embryos were analysed at stage 42. Image published in: Vacca B et al. (2016) © 2016. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |