利用斑馬魚建立心臟衰竭研究之動物模式:專一心臟troponin-C蛋白表現受抑制之斑馬魚心肌功能研究
Other Title
Conditional Antisense-Knockdown of Zebrafish Cardiac Troponin C as a
New Model for Dilated Cardiomyopathy
New Model for Dilated Cardiomyopathy
Date Issued
2005
Date
2005
Author(s)
何奕倫
DOI
932314B002219
Abstract
Background – Mutations of cardiac troponin C (cTnC) can cause dilated
cardiomyopathy in humans. We attempted to generate a zebrafish transgenic
line that enables us to produce the antinsense RNA strand of cTnC after
induction as a new animal model for studying dilated cardiomyopathy.
Methods and Results – We constructed plasmids that the reverse
tetracycline–controlled transactivator (rtTA) was driven by the cardiac myosin
light chain 2 promoter. This heart-specific rtTA bound another bidirectional
promoter to express the green fluorescence protein (GFP) reporter gene and
to transcribe the antisense RNA of cTnC in the presence of doxycycline (Dox).
After microinjection of these constructs into one-cell fertilized eggs, we
cultured, screened, and generated a transgenic line of zebrafish (CA17) which
enabled us to express GFP and to conditionally produce the antisense RNA of
cTnC in heart after induction. When 12-h postfertilization (hpf) embryos of F2
derived from CA17 were given Dox (10 µg/ml), the expression of endogenous
cTnC mRNA was not affected until 6 days postfertilization (dpf). The heart
rates of the embryos in CA17 were significantly slower than those of embryos
in the control T03 transgenic line (only expressing the heart-specific GFP) at 6
dpf (150±10 vs. 194±11 beats/min, p < 0.01) and 12 dpf (128±12 vs. 168±8 beats/min, p < 0.01). Moreover, the various values of cardiac chambers
measured in the induced F2 embryos from CA17 were significantly greater
than those of T03 embryos; while the ventricular ejection fraction of CA17 was
lower than that of the T03 both at 6 dpf (44%±4% vs. 50%±3%, p < 0.01) and
at 12 dpf (39%±5% vs. 52%±4%, p <0.01). In addition, asynchronized atrial
and ventricular contractions were noted in a few (1.3%) F2 embryos from the
induced CA17 transgenic fish. The mortality rate of F2 adult fish of the CA17
group was significantly higher (30% vs. 0%, p < 0.001) than that of F2 adults of
the T03 group after Dox induction.
Conclusions – Using conditional expression of antisense RNA of zebrafish
cTnC, we have created a new animal model with phenotypes simulating
dilated cardiomyopathy.
Subjects
Zebrafish
Cardiac troponin C
Antisense RNA
Dilated
cardiomyopathy
cardiomyopathy
SDGs
Publisher
臺北市：國立臺灣大學醫學院內科
Type
report
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