XB-IMG-172568
Xenbase Image ID: 172568
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Fig. 5.
Pitx2c is required for left liver lobe morphology. A Pitx2c morpholino was targeted to the left liver diverticulum, where pitx2c is normally expressed (see Fig. 4). Liver morphology was then analyzed in control and Pitx2-deficient embryos at NF 46 using in situ hybridization with a probe for fibrinogen (A-C). In control morpholino injected livers (CoMo, A), the right lobe (R, orange outline) is larger and more elongate than the left (L, yellow outline). In contrast, in Pitx2cMO injected livers (B-C), the left lobe is larger and more elongated, similar to control right side morphology (i.e., the liver exhibits right isomerism). Scale bars= 200 µm. D) Quantification of Normal, Left Isomerism (L-Iso), Right Isomerism (R-Iso), and LR Reversed (Rev) liver phenotypes (see cartoons) in Control and Pitx2cMo embryos. E) As expected in control embryos, the right lobe of the liver has a greater gross length/width ratio than the left lobe (NF 46). In contrast, in embryos injected with Pitx2cMO, the left lobe is larger, similar to a control right lobe. As expected, the dimensions of the right liver lobe are unaffected by the left-targeted Pitx2cMO. Error bars = S.E. Asterisks indicate p < 0.05 (*) or p < 0.0001 (**); NS, not significant. F-H) Similar results were obtained with CRISPR-Cas9-mediated Pitx2c loss of function. Liver lobe morphology was analyzed at NF 46 in embryos injected with synthetic Cas9 mRNA plus Tyr-gRNA (control; F) or plus pitx2c-gRNA (G-H) using in situ hybridization with a probe for fibrinogen. In control livers (F), the right lobe (R, orange outline) is larger and more elongate than the left (L, yellow outline). In contrast, in livers from pitx2c gRNA injected embryos (G-H), the left lobe is larger and more elongated, similar to control right side morphology (i.e., the liver exhibits right isomerism). Scale bars= 200 µm. Image published in: Womble M et al. (2018) Copyright © 2018. Image reproduced with permission of the Publisher, Elsevier B. V.
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