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Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
tek
|
|
laevis
|
NF stage 31
|
cardinal vein
,
anterior cardinal vein
,
blood vessel
,
vascular plexus
|
cfd
|
|
laevis
|
NF stage 31
|
cardinal vein
,
anterior cardinal vein
,
vascular plexus
,
vitelline vein
|
cfd
|
|
laevis
|
NF stage 32
|
cardinal vein
,
anterior cardinal vein
,
blood vessel
,
intersomitic vessel
,
vascular plexus
|
cfd
|
|
laevis
|
NF stage 35 and 36
|
cardinal vein
,
anterior cardinal vein
,
blood vessel
,
intersomitic vessel
,
vascular plexus
|
cfd
|
|
laevis
|
NF stage 40
|
cardinal vein
,
anterior cardinal vein
,
blood vessel
,
intersomitic vessel
,
vascular plexus
|
kdr
|
|
laevis
|
NF stage 31
|
cardinal vein
,
vein
,
blood vessel
,
intersomitic vessel
,
vascular plexus
|
aplnr
|
|
laevis
|
NF stage 31
|
cardinal vein
,
vein
,
aortic arch
,
blood vessel
,
intersomitic vessel
|
|
|
Fig. 3. XRASGRP2 depletion results in aberrant development of blood vessels. (A) Schematic model for the splice inhibition antisense morpholino oligo- nucleotides (S-MOs). The binding site of MO is represented by a bolded blue line. Arrows indicate the primers used in the RT-PCR to examine the efficacies of the S-MOs. (B) The control MO (c, 40 ng), aS- MO (aS, 40 ng), bS-MO (bS, 40 ng), and S- MO (S, 40 ng, comprising 20 ng aS-MO plus 20 ng bS-MO) were injected into 2- cell-stage embryos, and the embryos were analyzed by RT-PCR at stage 30. The presence of the 312-bp band indicates amplification of the normally spliced mRNA. The intensity of this band is re- duced in both the aS-MO-injected and bS- MO-injected embryos, as compared to the uninjected embryos and control MO- injected embryos, and this band is not detected for the S-MO-injected embryos. This indicates that the S-MO-injected embryos do not produce a functional XRASGRP2 protein. Sample without reverse transcriptase; n uninjected embryos. (C-F) Expression patterns of blood vessel marker genes. The 2-cell-stage embryos were injected with the control MO (40 ng) or S-MO (40 ng) into one blastomere (corresponding to the future right-hand side), and harvested at stage 31. The injected sides are indicated as [Inj(+)] and the uninjected sides are indi- cated as [Inj(-)]. The expression levels of Xflk-1 in the PCV (C, red arrows) and of Xmsr in the ISV (D, black arrows) are diminished in the S-MO-injected side. The expression levels of Xtie2 (E) and Ami (F) in the PCV (red arrows) and VVN (red arrowheads) are diminished in the S-MO-injected side. The expression level of Ami (F) is greatly reduced in the S-MO-in- jected side. No differences are seen in the control MO-injected embryos. (G) The expression of Ami in VVN is gradually mitigated in the S-MO-injected side. (H) Expression of globin T3 in the control MO-injected embryos. S-MO injection does not affect the level of globin T3 expression. |