Table 2.

Effect on D/V axis formation of antisense targeting of glce. Capped mRNA and glce-MOs were dissolved in Danieau's buffer and injected as 1–3 nl bolus into the yolk of one- to two-cell embryos as described in Table 1. Capped human GLCE-mRNA was generated from a full length cDNA clone. The embryos were classified according to the severity of the observed phenotype at 48 hpf based on a classification of the dorsalization severity observed in embryos that had only received MOs. Mildly dorsalized embryos show reduced ventral tail fin extension. Moderately dorsalized embryos display kinked tail, enlarged heart cavity and in some case the absence of atrioventricular boundary. Severely affected morphants show dramatic shortening and reduction of the body mass with tail coiling. For the rescue experiments, embryos were first injected with MO and after randomization half received the indicated amount of mRNA before reaching the 4-cell stage.

Treatment	Phenotype Severity
	No.	Wild	Mild	Moderate	Severe
		(%)	(%)	(%)	(%)
Uninjected	42	100	0	0	0
glce-A MO (4 ng)	62	35	46	19	0
glce-A MO (8 ng)	40	20	25	39	16
glce-A MO (16 ng)	28	12	19	31	38
glce-B MO (16 ng)	30	25	19	30	26
glce-A MO (8 ng) glce-B MO (8 ng)	30	15	10	30	45
glce-B MO (16 ng) glce-A mRNA (200 pg)	68	28	38	22	12
glce-B MO (16 ng) glce-B mRNA (200 pg)	30	32	45	15	8
glce-A MO (16 ng) Human GLCE mRNA (200 pg)	35	21	41	29	9