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The paracrine effect of exogenous growth hormone alleviates dysmorphogenesis caused by tbx5 deficiency in zebrafish (Danio rerio) embryos.
[holt-oram syndrome]
Dysmorphogenesis
and
multiple
organ
defects
are
well
known
in
zebrafish
(
Danio
rerio
)
embryos
with
T
-
box
transcription
factor
5
(
tbx
5
)
deficiencies
,
mimicking
human
Holt-
Oram
syndrome
.
Using
an
oligonucleotide-based
microarray
analysis
to
study
the
expression
of
special
genes
in
tbx
5
morphants
,
we
demonstrated
that
GH
and
some
GH-related
genes
were
markedly
downregulated
.
Zebrafish
embryos
microinjected
with
tbx
5
-
morpholino
(
MO
)
antisense
RNA
and
mismatched
antisense
RNA
in
the
1
-
cell
stage
served
as
controls
,
while
zebrafish
embryos
co
-injected
with
exogenous
growth
hormone
(
GH
)
concomitant
with
tbx
5
-
MO
comprised
the
treatment
group
.
The
attenuating
effects
of
GH
in
tbx
5
-
MO
knockdown
embryos
were
quantified
and
observed
at
24
,
30
,
48
,
72
,
and
96
h
post-fertilization
.
Though
the
understanding
of
mechanisms
involving
GH
in
the
tbx
5
functioning
complex
is
limited
,
exogenous
GH
supplied
to
tbx
5
knockdown
zebrafish
embryos
is
able
to
enhance
the
expression
of
downstream
mediators
in
the
GH
and
insulin
-like
growth
factor
(
IGF
)
-
1
pathway
,
including
igf
1
,
ghra
,
and
ghrb
,
and
signal
transductors
(
erk
1
,
akt
2
)
,
and
eventually
to
correct
dysmorphogenesis
in
various
organs
including
the
heart
and
pectoral
fins
.
Supplementary
GH
also
reduced
apoptosis
as
determined
by
a
TUNEL
assay
and
decreased
the
expression
of
apoptosis-related
genes
and
proteins
(
bcl
2
and
bad
)
according
to
semiquantitative
reverse-transcription
polymerase
chain
reaction
and
immunohistochemical
analysis
,
respectively
,
as
well
as
improving
cell
cycle-related
genes
(
p
27
and
cdk
2
)
and
cardiomyogenetic
genes
(
amhc
,
vmhc
,
and
cmlc
2
)
.
Based
on
our
results
,
tbx
5
knockdown
causes
a
pseudo
GH
deficiency
in
zebrafish
during
early
embryonic
stages
,
and
supplementation
of
exogenous
GH
can
partially
restore
dysmorphogenesis
,
apoptosis
,
cell
growth
inhibition
,
and
abnormal
cardiomyogenesis
in
tbx
5
knockdown
zebrafish
in
a
paracrine
manner
.
Diseases
Validation
Diseases presenting
"growth hormone"
symptom
achondroplasia
aniridia
aromatase deficiency
cohen syndrome
dentinogenesis imperfecta
holt-oram syndrome
megacystis-microcolon-intestinal hypoperistalsis syndrome
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