Abstract
BackgroundSlc39a7/Zip7, also known as Ke4, is a member of solute carrier family 39 (Slc39a) and plays a critical role in regulating cell growth and death. Because the function of Zip7 in vivo was unclear, the present study investigated the function of zip7 in vertebrate development and zinc metabolism using zebrafish as a model organism.Principal FindingUsing real-time PCR to determine the gene expression pattern of zip7 during zebrafish development, we found that zip7 mRNA is expressed throughout embryonic development and into maturity. Interestingly, whole mount in situ hybridization revealed that while zip7 mRNA is ubiquitously expressed until 12 hours post-fertilization (hpf); at 24 hpf and beyond, zip7 mRNA was specifically detected only in eyes. Morpholino-antisense (MO) gene knockdown assay revealed that downregulation of zip7 expression resulted in several morphological defects in zebrafish including decreased head size, smaller eyes, shorter palates, and shorter and curved spinal cords. Analysis by synchrotron radiation X-ray fluorescence (SR-XRF) showed reduced concentrations of zinc in brain, eyes, and gills of zip7-MO-injected embryos. Furthermore, incubation of the zip7 knockdown embryos in a zinc-supplemented solution was able to rescue the MO-induced morphological defects.SignificanceOur data suggest that zip7 is required for eye, brain, and skeleton formation during early embryonic development in zebrafish. Moreover, zinc supplementation can partially rescue defects resulting from zip7 gene knockdown. Taken together, our data provide critical insight into a novel function of zip7 in development and zinc homeostasis in vivo in zebrafish.
Highlights
Zinc is an essential trace element required for DNA synthesis, cell division, regulation of transcription, and protein synthesis
The expression pattern of zip7 mRNA during zebrafish embryogenesis
We used whole-mount in situ hybridization to determine the spatial distribution of zip7 gene expression during zebrafish embryogenesis and found that at early stages of somitogenesis, zip7 mRNA was ubiquitously expressed (Figure 1A)
Summary
Zinc is an essential trace element required for DNA synthesis, cell division, regulation of transcription, and protein synthesis. 2000 enzymes use zinc as a catalytic cofactor [1], and zinc binding motifs are found in up to 10% of the proteins encoded by the human genome [2] including zinc-fingercontaining proteins, the most abundant protein superfamily in the mammalian genome In this regard, zinc is an essential cofactor required for the activity of numerous proteins involved in cellular signaling pathways and biological processes including growth factors, cytokines, receptors, enzymes, and transcription factors [3,4,5,6]. Slc39a7/Zip, known as Ke4, is a member of solute carrier family 39 (Slc39a) and plays a critical role in regulating cell growth and death. Incubation of the zip knockdown embryos in a zinc-supplemented solution was able to rescue the MO-induced morphological defects
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