| |
Figure 3. Impairment of eye development upon Xdhcr7 knockdown. a-j rdquo : Microinjection of Xdhcr7-morpholino (1 pmol) in one blastomere of two-cell-stage embryos. In combination, 200 pg of beta -galactosidase mRNA was injected as linage tracer (blue staining). k-k rdquo ': Microinjection of Xdhcr7-morpholino (0.5 pmol) in two blastomeres of two-cell-stage embryos. l: Microinjection of Xdhcr7-morpholino (1 pmol) in two blastomeres of two-cell-stage embryos (a, c, e) Anterior view (dorsal up); dashed lines indicate the midline. b, b rdquo , d, d rdquo , f, f rdquo : Lateral view (dorsal up). k rdquo : Ventral view (anterior down). b', d', f': Frontal view (dorsal up). g-l: Dorsal view (anterior down). f rdquo ', g', h', j', j rdquo , k rdquo ': Transversal sections. b rdquo ', d rdquo ': Horizontal sections; dashed lines indicate the midline. a-h': WMISH analysis of Xrx1/Xen2/Xkrox20 (a-b rdquo '), Xsix3 (c-d rdquo '), Xn-tubulin (e-f rdquo '), Xgsh1 (g, g'), and Xnkx2.2 (h, h') expression. i-l: Eye phenotypes. a: Suppression of Xkrox20 (r3 and r5; 40%, n = 25) and Xen2 (mhb; 40%, n = 25) expression. a,c: Expansion of Xrx1 (48%, n = 25) and Xsix3 (56%, n = 25) expression at neurula stage. b-b rdquo ': Reduction of Xrx1 (56%, n = 32), Xen2 (53%, n = 32), and Xkrox20 (65%, n = 32) expression at tailbud stage. The expression domain of Xrx1 is shifted towards the midline (b rdquo '). d-d rdquo ': Severe reduction of Xsix3 expression (67%, n = 28). Retinal Xsix3 expression is almost linked to the brain (d rdquo '). e: Severe reduction of Xn-tubulin expression at neurula stage (72%, n = 29). Note the reduced expression in both the neural plate and the trigeminal placode. f-f rdquo ': Severe reduction of Xn-tubulin expression at tailbud stage (57%, n = 35). Xn-tubulin expression is almost absent in the neural tube (compare injected and non-injected side in f rdquo '). g-h': Suppression of Xgsh1 (63%, n = 30) and Xnkx2.2 (59%, n = 32) expression. The expression domain of Xnkx2.2 is slightly shifted ventrally. i-i rdquo : Small eye (60%, n = 300). The small eye is closer to the brain and displays a coloboma (i rdquo ). j-j rdquo : Severely reduced eye formation (33%, n = 300). The retinal pigmented epithelium is almost absent in the eye of the injected side (white arrowhead in j rdquo ). k-k rdquo ': Mild eyes phenotype induced by injection of 0.5 pmol Xdhcr7-morpholino in two cells of two-cell stage. l: Cyclopic eye induced by bilateral injection of 1 pmol Xdhcr7-morpholino. cp, cranial placode; cs, control side; ey, eye; is, injected side; le, lens; mhb, midbrain-hindbrain boundary; r3, rhombomere 3; r5, rhombomere 5; rpe, retinal pigmented epithelium; tp, trigeminal placode. |
