- Title
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A Functional Assay for Sick Sinus Syndrome Genetic Variants
- Authors
- Jou, C.J., Arrington, C.B., Barnett, S., Shen, J., Cho, S., Sheng, X., McCullagh, P.C., Bowles, N.E., Pribble, C.M., Saarel, E.V., Pilcher, T.A., Etheridge, S.P., Tristani-Firouzi, M.
- Source
- Full text @ Cell Physiol. Biochem.
Embryonic hnc4 expression: whole-mount in situ hybridization reveals hcn4 expression in the heart (arrow) at (A) 18 somites, (B) 24 hpf and (C) 48 hpf. To explore the mechanism by which hcn4 functions in zebrafish, we bathed whole embryos in 4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino) pyrimidinium chloride (ZD-7288), a nonselective hcn blocker, and assayed the heart rate at 48 hpf. We found that embryos bathed in 1 mM ZD-7288 had significantly slower heart rates than untreated embryos (Fig. 2A). However, cardiac pauses were not observed in embryos exposed to ZD-7288. EXPRESSION / LABELING:
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ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions. PHENOTYPE:
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ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions. PHENOTYPE:
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Specificity of hcn4 MO. Retention of intron 3 with injection of hcn4 splice-blocking morpholino. RNA from uninjected and morphant embryos was analyzed by RT-PCR. Primers designed to amplify the exon3-exon4 boundary produced the expected PCR product in uninjected embryos (lane 2). The PCR product is too large to amplify in morphants due to inclusion of intron 3 (lane 3). To prove inclusion of intron 3 in morphant hcn4 mRNA, primers were designed to amplify the exon3-intron3 and intron3-exon4 boundaries. Products of the expected size were found in morphants (lanes 4 and 5, respectively). |