کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10926333 | 1091815 | 2013 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Transgenic zebrafish for ratiometric imaging of cytosolic and mitochondrial Ca2+ response in teleost embryo
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیولوژی سلول
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چکیده انگلیسی
Intracellular Ca2+ imaging has widely been used to visualize intracellular signals, but the application in an intact animal is still limited due to difficulty of the indicator loading. In addition, the motion of the living animal produces artifacts. To investigate Ca2+ signaling at early embryonic stage, we established transgenic zebrafish line expressing a genetically encoded Ca2+ indicator, cameleon YC2.60, driven by a constitutively active promoter, hspa8. Although the embryo dynamically changes its morphology, the motion artifact could be canceled out by taking the advantage of YC2.60 as a ratiometric indicator. The transgenic zebrafish was used to visualize the propagation of cytosolic Ca2+ during the early embryonic stage upon fertilization and along cleavage furrow, and the rise in Ca2+ in the myocytes contracting spontaneously in the embryo. We also established a transgenic zebrafish line expressing YC2.60 targeted to the mitochondria. The rise in mitochondrial Ca2+ was rather sustained (â2Â min), which is consistent with the requirement of ATP refilling since the mitochondrial Ca2+ upregulates rate-limiting enzymes of Krebs cycle. This is in contrast with the transient rise in the cytosol Ca2+ that directly evokes the muscle contraction. These transgenic zebrafish lines are expected to serve as useful tools further Ca2+ imaging in vivo.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Cell Calcium - Volume 54, Issue 3, September 2013, Pages 236-245
Journal: Cell Calcium - Volume 54, Issue 3, September 2013, Pages 236-245
نویسندگان
Hideaki Mizuno, Takayuki Sassa, Shin-ichi Higashijima, Hitoshi Okamoto, Atsushi Miyawaki,