Functional analysis of KCNH2 gene mutations of type 2 long QT syndrome in larval zebrafish using microscopy and electrocardiography View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2018-07-25

AUTHORS

Yoshihiro Tanaka, Kenshi Hayashi, Noboru Fujino, Tetsuo Konno, Hayato Tada, Chiaki Nakanishi, Akihiko Hodatsu, Toyonobu Tsuda, Yoji Nagata, Ryota Teramoto, Shohei Yoshida, Akihiro Nomura, Masa-aki Kawashiri, Masakazu Yamagishi

ABSTRACT

Heterologous expression systems play a vital role in the characterization of potassium voltage-gated channel subfamily H member 2 (KCNH2) gene mutations, such as E637K which is associated with long QT syndrome type 2 (LQT2). In vivo assays using zebrafish provide a means for testing genetic variants of cardiac disease; however, limited information on the role of the E637K mutation is available from in vivo systems and their utility has yet to be fully exploited in the context of LQT2. We sought to evaluate the ability of the E637K mutant channel to restore normal repolarization in larval zebrafish with a human KCNH2 orthologue, kcnh2a-knockdown. A morpholino (MO) targeting kcnh2a was injected alone or with wild type (WT) or E637K KCNH2 cRNA into zebrafish embryos at the 1–2 cell stage. Cardiac repolarization phenotypes were screened using light microscopy and the QT interval was measured by single lead electrocardiograph (ECG) analysis at 72-h post-fertilization. In the MO alone group, 17% of zebrafish had a normal phenotype; this rate increased to 60% in the WT KCNH2 cRNA injected zebrafish and to 35% in the E637K injected zebrafish. The ECG of larval zebrafish revealed that QTc was significantly prolonged in the MO alone group compared to the control group. Co-injection of WT KCNH2 cRNA shortened the QTc interval, however, that of the E637K did not. We suggest that this in vivo cardiac assay using microscopy and ECG in larval zebrafish offers a reliable approach for risk discrimination of KCNH2 mutations. More... »

PAGES

159-166

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00380-018-1231-4

DOI

http://dx.doi.org/10.1007/s00380-018-1231-4

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1105808512

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30047011


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24 schema:description Heterologous expression systems play a vital role in the characterization of potassium voltage-gated channel subfamily H member 2 (KCNH2) gene mutations, such as E637K which is associated with long QT syndrome type 2 (LQT2). In vivo assays using zebrafish provide a means for testing genetic variants of cardiac disease; however, limited information on the role of the E637K mutation is available from in vivo systems and their utility has yet to be fully exploited in the context of LQT2. We sought to evaluate the ability of the E637K mutant channel to restore normal repolarization in larval zebrafish with a human KCNH2 orthologue, kcnh2a-knockdown. A morpholino (MO) targeting kcnh2a was injected alone or with wild type (WT) or E637K KCNH2 cRNA into zebrafish embryos at the 1–2 cell stage. Cardiac repolarization phenotypes were screened using light microscopy and the QT interval was measured by single lead electrocardiograph (ECG) analysis at 72-h post-fertilization. In the MO alone group, 17% of zebrafish had a normal phenotype; this rate increased to 60% in the WT KCNH2 cRNA injected zebrafish and to 35% in the E637K injected zebrafish. The ECG of larval zebrafish revealed that QTc was significantly prolonged in the MO alone group compared to the control group. Co-injection of WT KCNH2 cRNA shortened the QTc interval, however, that of the E637K did not. We suggest that this in vivo cardiac assay using microscopy and ECG in larval zebrafish offers a reliable approach for risk discrimination of KCNH2 mutations.
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31 K mutation
32 KCNH2 gene mutations
33 KCNH2 mutations
34 Mo
35 QT interval
36 QT syndrome
37 QT syndrome type 2
38 QTc
39 QTc interval
40 ability
41 analysis
42 approach
43 assays
44 cRNA
45 cardiac disease
46 cell stage
47 channels
48 characterization
49 context
50 control group
51 discrimination
52 disease
53 electrocardiograph (ECG) analysis
54 electrocardiography
55 embryos
56 expression system
57 functional analysis
58 gene mutations
59 genetic variants
60 group
61 heterologous expression system
62 information
63 interval
64 kcnh2a
65 larval zebrafish
66 light microscopy
67 limited information
68 long QT syndrome
69 long QT syndrome type 2
70 means
71 microscopy
72 mutant channels
73 mutations
74 normal phenotype
75 normal repolarization
76 orthologues
77 phenotype
78 potassium voltage-gated channel
79 rate
80 reliable approach
81 repolarization
82 risk discrimination
83 role
84 stage
85 syndrome
86 system
87 type 2
88 type 2 long QT syndrome
89 types
90 utility
91 variants
92 vital role
93 vivo
94 vivo assays
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