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Xenbase Image ID: 49651

Fig. 2. Depletion of Coco protein randomizes the left right axis. (A) Alignment of the Coco and Coco2 sequences. (B) Design of Coco MO1 and 2. A mix of both was used. (C) Coco MO specifically inhibits translation of injected and endogenous wild-type Coco RNA but not of a 5′ mutated RNA (Mut Coco). Western blots. Upper panel. 100 pg Coco or Mut Coco RNA were injected in the animal pole of each blastomere at the two-cell stage, alone (lanes 2, 5) or with 20 ng of either Coco MO (lanes 3, 6) or Coco Mut MO (lane 4). Animal caps were collected at stage 10. Coco protein was detected with anti-Coco antibody (1:10,000). Lower panel. Expression of endogenous Coco protein is inhibited by Coco MOs. The diagram represents a neurula-stage embryos, dorsal view, with the dissected dorsal posterior fragment boxed. Each lane contains 100 dissected dorsal posterior fragments (stage 18) from embryos injected dorsally with 3 ng control MO or 3 ng Coco MO. Positive control was overexpressed Coco FLAG RNA. Polyclonal antibodies for Coco (1:500) and β-catenin (1:10,000) were used. (D) Stage 46 embryos, dorsal (whole, D, H) and ventral (details) views. (D) Control embryos. (H) Embryos injected at four-cell stage in the dorsal right blastomere with 3 ng Coco MO. (E, I) Immunohistochemistry (peroxidase), (F, J) immunofluorescence for cardiac troponin. V: ventricle; A: atrium; Ao: aorta. Curved arrows indicate the direction of the outflow tract and of the gut looping, straight arrows in panels F and J indicate the antero-posterior axis. Panel I is a complete inversion (Situs inversus), panel K is an isolated gut inversion and panel L is an isolated heart inversion (Situs ambiguus). Hearts are contoured in panels G1, K1 and L1, and G2, K2, and L2 show the position of the gall bladder by UV autofluorescence in the same embryos.

Image published in: Vonica A and Brivanlou AH (2007)

Copyright © 2007. Image reproduced with permission of the Publisher, Elsevier B. V.

tnnt2.LCT3X.laevisThroughout NF stage 46heart

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