|
Gene |
Clone |
Species |
Stages |
Anatomy |
pcna.S
|
|
laevis
|
NF stage 14
to
NF stage 15
|
neural plate
,
anterior neural fold
,
preplacodal ectoderm
|
pcna.S
|
|
laevis
|
NF stage 19
to
NF stage 20
|
brain
,
branchial crest
,
eye primordium
,
cranial neural crest
,
neural tube
|
pcna.S
|
|
laevis
|
NF stage 25
to
NF stage 26
|
brain
,
spinal cord
,
eye
,
pharyngeal region
|
pcna.S
|
|
laevis
|
NF stage 37 and 38
|
olfactory placode
,
otic vesicle
,
brain
,
forebrain
,
midbrain-hindbrain boundary
,
[+]
|
|
|
Fig. 5. Expression pattern of PCNA at several developmental stages during MeHgCl solution exposure, as detected by ISH. Stages 14–15 (A–F), stages 19–20 (G–L), and stages 25–26 (M–R) embryos are seen from a dorsal view. Stages 37–38 embryos (S–X) are shown from a dorsal (left) and lateral (right) view, except for 200 μg/l, which is a featureless ball of cells. All embryos shown are representative near the average phenotype score from exposure at each dose. For all images, anterior is on the right. Dark purple-blue areas indicate positive PCNA expression. N = 2 experiments, with the total number of evaluated embryos shown at the lower right of each image. The number of embryos out of the total matching each image is also shown. (Y) qRT-PCR results of relative PCNA expression in stage 37 embryos at each MeHgCl dose. Values for the δδCт score in each condition have been normalized to the expression level of the 0 μg/l control and are shown using 2− δδCт scale. N = 1 biological experiment pooled from 10 embryos, and 3 technical replicates, for each dose. Scale bars represent 1 mm. |
|
|
|
Fig. 6. Transverse histological cryosections through stage 37 embryos stained for PCNA expression (blue regions) by ISH. Forebrain, midbrain, hindbrain, and spinal cord sections of control (A–D) and MeHgCl solution exposed (E–H) embryos are shown oriented dorsal on top. Two of the four MeHgCl exposed embryos displayed abnormally reduced or absent forebrain structures as seen in (E). N = 2 experiments, with 4 embryos sectioned from each condition. (I) Neural cell counts using DAPI stain compared between unexposed and MeHgCl exposed embryos averaged over each CNS region. N = 4 experiments, with 7 embryos sectioned from 0 μg/l and 8 from 100 μg/l total. Error bars represent standard deviation. *p = 0.039, **p = 0.0056, ns = not significant as determined by Student's t-test for each region. Abbreviations: e, eye; fb, forebrain; hb, hindbrain; mb, midbrain; olf, olfactory; ot, otic vesicle; pn, pronephros; sc, spinal cord. Scale bar represents 150 μm. |