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Table 1 Effect of the ectopic expression of Glce on Bmp2b ventralizing activity. Capped glce and bmp2b mRNAs were generated by reverse transcription from full length cDNA clones. After extraction in phenol/chloroform and precipitation in isopropanol, mRNA was dissolved in Danieau's buffer and the concentration assessed by UV reading (260 nm). Embryos at one or two cell stage were injected with 1–3 nl of mRNAs to achieve the indicated dose. Each injection session consisted of 2–3 treatment groups of 30 embryos each and several experiments were performed to reach the sample number indicated. Embryos with increasing degree of ventralization were ranked according to previously established criterias [40,41]. Ventralized V1 embryos show reduced eye size and shorter body. In addition to these abnormalities V2 embryos display abnormal notocord, reduced anterior somites, and enlarged blood islands. Ventralized V3 embryos have little or no head structures and no notochord. V4 embryos display gross body abnormalities and lack of anterior structures.

From: D-glucuronyl C5-epimerase acts in dorso-ventral axis formation in zebrafish

Injected mRNA

Strength of Ventralization

 

No.

Wild Type

V1

V2

V3

V4

  

(%)

(%)

(%)

(%)

(%)

Uninjected

120

100

0

0

0

0

glce-A (250 pg)

60

65

23

10

2

0

glce-B (250 pg)

60

54

16

28

2

0

glce-A (125 pg)

glce-B (125 pg)

30

60

7

23

10

0

glce-A (250 pg)

glce-B (250 pg)

30

37

6

10

47

0

glce-A (500 pg)

glce-B (500 pg)

60

26

0

38

35

0

bmp2b (20 pg)

50

0

26

37

26

11

bmp2b (20 pg)

glce-A (250 pg)

60

0

0

15

45

40

bmp2b (20 pg)

glce-B (250 pg)

30

0

0

26

44

30