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Neurulation and neurite extension require the zinc transporter ZIP12 (slc39a12).

ABSTRACT: Zn(2+) is required for many aspects of neuronal structure and function. However, the regulation of Zn(2+) in the nervous system remains poorly understood. Systematic analysis of tissue-profiling microarray data showed that the zinc transporter ZIP12 (slc39a12) is highly expressed in the human brain. In the work reported here, we confirmed that ZIP12 is a Zn(2+) uptake transporter with a conserved pattern of high expression in the mouse and Xenopus nervous system. Mouse neurons and Neuro-2a cells produce fewer and shorter neurites after ZIP12 knockdown without affecting cell viability. Zn(2+) chelation or loading in cells to alter Zn(2+) availability respectively mimicked or reduced the effects of ZIP12 knockdown on neurite outgrowth. ZIP12 knockdown reduces cAMP response element-binding protein activation and phosphorylation at serine 133, which is a critical pathway for neuronal differentiation. Constitutive cAMP response element-binding protein activation restores impairments in neurite outgrowth caused by Zn(2+) chelation or ZIP12 knockdown. ZIP12 knockdown also reduces tubulin polymerization and increases sensitivity to nocodazole following neurite outgrowth. We find that ZIP12 is expressed during neurulation and early nervous system development in Xenopus tropicalis, where ZIP12 antisense morpholino knockdown impairs neural tube closure and arrests development during neurulation with concomitant reduction in tubulin polymerization in the neural plate. This study identifies a Zn(2+) transporter that is specifically required for nervous system development and provides tangible links between Zn(2+), neurulation, and neuronal differentiation.

SUBMITTER: Chowanadisai W

PROVIDER: S-EPMC3683776 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

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Neurulation and neurite extension require the zinc transporter ZIP12 (slc39a12).

Chowanadisai Winyoo W Graham David M DM Keen Carl L CL Rucker Robert B RB Messerli Mark A MA

Proceedings of the National Academy of Sciences of the United States of America 20130528 24

Zn(2+) is required for many aspects of neuronal structure and function. However, the regulation of Zn(2+) in the nervous system remains poorly understood. Systematic analysis of tissue-profiling microarray data showed that the zinc transporter ZIP12 (slc39a12) is highly expressed in the human brain. In the work reported here, we confirmed that ZIP12 is a Zn(2+) uptake transporter with a conserved pattern of high expression in the mouse and Xenopus nervous system. Mouse neurons and Neuro-2a cells ...[more]

PMID: 23716681

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Project description:Abstract Objectives Zinc is important for brain function. The SLC39A12 gene encodes ZIP12, which is a zinc transport protein highly expressed in the brain. Our goal is to determine whether human slc39a12 coding polymorphisms associated with differences in susceptibility-weighted brain MRI (swMRI) patterns affect zinc transport ability. Methods We used summary statistics for a genome-wide association study (GWAS) on swMRI patterns for caudate, putamen, and pallidum of 7778 human subjects aged 40–69 years from the UK BioBank. Human SLC39A12 coding polymorphisms rs10764176 and rs72778328 were associated with a reduction in relaxation spin time during swMRI in the putamen or pallidum but not caudate and met a significance cutoff of P < 5.0 × 10–8. Haploblock determination and linkage disequilibrium (LD) between the variants was determined by SNPclip, LDmatrix, and LDpair software and the European 1000 genomes LD reference panel. The effects of the ZIP12 polymorphisms on cell zinc status and zinc uptake were measured in Chinese hamster ovary (CHO) cells transfected with the reference ZIP12 version or coding variants. Cell zinc status was assessed after 18 hours by metal-response element reporter activation (n = 8). Cell zinc uptake activity was measured by stable isotope Zinc-70 uptake (n = 6) relative to endogenous Zinc-68 content using inductively coupled plasma mass spectrometry (ICP-MS). Statistical comparisons were performed using ANOVA. Significance was set at P < 0.05. Results Variants rs10764176 and rs72778328 are present on independent haploblocks and are in linkage equilibrium with each other. Variants rs10764176 and rs72778328 have a reduction in MRE reporter activation (41.7% [P < 0.05] and 55.6% [P < 0.01], respectively) and in exogenous Zinc-70 uptake after 20 minutes (37.6% [P < 0.001] and 54.2% [P < 0.001], respectively) compared to the reference ZIP12. Conclusions Variants rs10764176 and rs72778328 are on distinct haploblocks. These ZIP12 coding variants that are associated with altered brain swMRI patterns also have reduced zinc uptake activity. Funding Sources This work was funded by grants from the Oklahoma Center for the Advancement of Science and Technology and the Oklahoma Agricultural Experiment Station.

Project description:BackgroundOverwhelming evidences suggest oxidative stress is a major cause of sperm dysfunction and male infertility. Zinc is an important non-enzymatic antioxidant with a wide range of biological functions and plays a significant role in preserving male fertility. Notably, zinc trafficking through the cellular and intracellular membrane is mediated by specific families of zinc transporters, i.e., SLC39s/ZIPs and SLC30s/ZnTs. However, their expression and function were rarely evaluated in the male germ cells. The aim of this study is to determine and characterize the crucial zinc transporter responsible for the maintenance of spermatogenesis.MethodsThe expression patterns of all 14 ZIP members were characterized in the mouse testis. qRT-PCR, immunoblot and immunohistochemistry analyses evaluated the ZIP12 gene and protein expression levels. The role of ZIP12 expression was evaluated in suppressing the sperm quality induced by exposure to an oxidative stress in a spermatogonia C18-4 cell line. Zip12 RNAi transfection was performed to determine if its downregulation altered cell viability and apoptosis in this cell line. An obese mouse model fed a high-fat-diet was employed to determine if there is a correlation between changes in the ZIP12 expression level and sperm quality.ResultsThe ZIP12 mRNA and protein expression levels were higher than those of other ZIP family members in both the mouse testis and other tissues. Importantly, the ZIP12 expression levels were very significantly higher in both mice and human spermatogonia and spermatozoa. Moreover, the testicular ZIP12 expression levels significantly decreased in obese mice, which was associated with reduced sperm zinc content, excessive sperm ROS generation, poor sperm quality and male subfertility. Similarly, exposure to an oxidative stress induced significant declines in the ZIP12 expression level in C18-4 cells. Knockdown of ZIP12 expression mediated by transfection of a ZIP12 siRNA reduced both the zinc content and viability whereas apoptotic activity increased in the C18-4 cell line.ConclusionsThe testicular zinc transporter ZIP12 expression levels especially in spermatogonia and spermatozoa are higher than in other tissues. ZIP12 may play a key role in maintaining intracellular zinc content at levels that reduce the inhibitory effects of rises in oxidative stress on spermatogonia and spermatozoa viability during spermatogenesis which help counteract declines in male fertility.

Dataset Information

Neurulation and neurite extension require the zinc transporter ZIP12 (slc39a12).

Publications

Neurulation and neurite extension require the zinc transporter ZIP12 (slc39a12).

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