XB-IMG-156909
Xenbase Image ID: 156909
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Fig. 2.
Alcam deficiency results in abnormal eye development. A. The spatial expression pattern of alcam by WMISH shows alcam expression in the developing eye (white arrowheads) and the neural tube (black arrowheads) at indicated stages. At stage 42, alcam RNA was predominantly found in the ganglion cell layer (lower left panel), the lens epithelium (LE) and the cornea epithelium (CE) as shown by WMISH. Antibody staining demonstrated that the Alcam protein is mainly localized in the ciliary marginal zone (CMZ) and inner synaptic and nuclear layer of the retina (lower right panel). B. Loss of Alcam led to smaller and deformed eyes on the injected side (white arrows) in comparison to the un-injected side or Control MO injected embryos. Furthermore, Alcam depletion results in a disorganized retinal lamination as well as absent RPE (red arrows). Representative embryos are shown. Scale bar (sections): 100 µm. C. Quantitative representation of the data shown in B. Loss of Alcam led to disturbed eye morphology in a MO-dose dependent manner. Co-injection of alcam RNA significantly rescued the Alcam MO-induced eye phenotype. D. Detailed views of Control MO and Alcam MO injected embryos. Dashed red lines indicate measured eye areas. Red lines indicate the eye fissure angle (°fis.) measured. Quantitative representations of eye area and eye fissure angle (°fis.) measurements are given. E. Cross sections of Alcam-depleted embryos after WMISH. In contrast to the un-injected side, loss of Alcam led to a disorganization of the retinal cell layers due to ectopic formation of rosette-like structures of photoreceptor cells (red arrowheads). The ratios of Alcam MO injected embryos showing retinal lamination defects versus all investigated embryos are indicated in each marker gene picture respectively. Scale bar: 100 µm. F. Lens specific markers celf1 and cryba1 are shown by WMISH. Marker gene expression was unaffected upon Alcam depletion although some lenses were smaller (the ratios of Alcam MO injected embryos showing smaller lenses versus all investigated embryos are indicated). Scale bar: 100 µm. Abbreviations: CE, cornea epithelium; CMZ, ciliary marginal zone; GCL, ganglion cell layer; INL, inner nuclear layer; inj., injected; ISL, inner synaptic layer; LE, lens epithelium; n, number of independent experiments; N, total number of analyzed embryos; ONL, outer nuclear layer; RPE, retinal pigmented epithelium; st, stage. Error bars indicate standard error of the mean (s.e.m.). ** p≤0.01, *** p≤0.001, **** p≤0.0001. p values were calculated by non-parametric Mann–Whitney rank sum test. Image published in: Seigfried FA et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V.
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