Expression pattern and functional analysis of fundc1 in rare minnow ( Gobiocypris rarus )

Xu, Gongyu; Li, Zhenzhen; Xiao, Jinwen; Li, Fangqing; Ye, Weiyuan; Zhao, Haobin; Zhou, Qingchun

doi:10.1016/j.gene.2017.05.015

Cited by 3 publications

(3 citation statements)

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“…Zebrafish Fundc1 is involved in mitophagy, as described above. In this study, Drfundc1 knockdown caused severe defects in the body axis, such as midline bifurcation and headlessness, which had been previously observed in rare minnows 25 . Co-injection of Drfundc1 mRNA repaired these defects, although Drfundc1 ’s expression level did not go back to typical levels.…”

Section: Discussionsupporting

confidence: 74%

“…Whole mount in situ hybridization (WISH) was carried out on zebrafish embryos following the protocol reported previously 63 . Sense and antisense digoxigenin (DIG)-labeled RNA riboprobes were produced, as reported previously 25 . Embryos older than 20 hpf were digested with proteinase K and then hybridized with appropriate riboprobes at 70 °C for 12–16 h. After thoroughly washing and blocking, embryos were incubated with anti-DIG antibodies which conjugated with alkaline phosphatase.…”

Section: Methodsmentioning

confidence: 99%

“…Authors have previously found that knockdown of fundc1 causes severe defects in the body axis of a rare minnow ( Gobiocypris rarus ) 25 . However, the role of FUNDC1 in embryogenesis remains unclear.…”

Section: Introductionmentioning

confidence: 99%

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Fundc1 is necessary for proper body axis formation during embryogenesis in zebrafish

Shen

Nibona

et al. 2019

Sci Rep

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FUN14 domain-containing protein 1 (FUNDC1) is a mitochondrial outer membrane protein which is responsible for hypoxia-induced mitophagy in mammalian cells. Knockdown of fundc1 is known to cause severe defects in the body axis of a rare minnow. To understand the role of Fundc1 in embryogenesis, we used zebrafish in this study. We used bioimaging to locate zebrafish Fundc1 (DrFundc1) with MitoTracker, a marker of mitochondria, and/or CellLight Lysosomes-GFP, a label of lysosomes, in the transfected ovary cells of grass carp. The use of Western blotting detected DrFundc1 as a component of mitochondrial proteins with endogenous COX IV, LC3B, and FUNDC1 in transgenic human embryonic kidney 293 T cells. DrFundc1 induced LC3B activation. The ectopic expression of Drfundc1 caused cell death and apoptosis as well as impairing cell proliferation in the 293 T cell line, as detected by Trypan blue, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and incorporation of BrdU. DrFundc1 up-regulated expression of both autophagy- and apoptosis-related genes, including ATG5, ATG7, LC3B, BECLIN1, and BAX in transgenic 293 T cells. A knockdown of Drfundc1 using short hairpin RNA (shRNA) led to midline bifurcation with two notochords and two spinal cords in zebrafish embryos. Co-injection of Drfundc1 mRNA repaired defects resulting from shRNA. Knockdown of Drfundc1 resulted in up- or down-regulation of genes related to autophagy and apoptosis, as well as decreased expression of neural genes such as cyclinD1, pax2a, opl, and neuroD1. In summary, DrFundc1 is a mitochondrial protein which is involved in mitophagy and is critical for typical body axis development in zebrafish.

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Section: Discussionsupporting

confidence: 74%

Section: Methodsmentioning

confidence: 99%