Altered Expression of 14-3-3ζ Protein in Spinal Cords of Rat Fetuses with Spina Bifida Aperta

Abstract

BackgroundA large number of studies have confirmed that excessive apoptosis is one of the reasons for deficient neuronal function in neural tube defects (NTDs). A previous study from our laboratory used 2-D gel electrophoresis to demonstrate that 14-3-3ζ expression was low in the spinal cords of rat fetuses with spina bifida aperta at embryonic day (E) 17. As a member of the 14-3-3 protein family, 14-3-3ζ plays a crucial role in the determination of cell fate and anti-apoptotic activity. However, neither the expression of 14-3-3ζ in defective spinal cords, nor the correlation between 14-3-3ζ and excessive apoptosis in NTDs has been fully confirmed.Methodology/Principal FindingsWe used immunoblotting and quantitative real-time PCR (qRT-PCR) to quantify the expression of 14-3-3ζ and double immunofluorescence to visualize 14-3-3ζ and apoptosis. We found that, compared with controls, 14-3-3ζ was down-regulated in spina bifida between E12 and E15. Excessive apoptotic cells and low expression of 14-3-3ζ were observed in the dorsal region of spinal cords with spina bifida during the same time period. To initially explore the molecular mechanisms of apoptosis in NTDs, we investigated the expression of microRNA-7 (miR-7), microRNA-375 (miR-375) and microRNA-451 (miR-451), which are known to down-regulate 14-3-3ζ in several different cell types. We also investigated the expression of p53, a molecule that is downstream of 14-3-3ζ and can be down-regulated by it. We discovered that, in contrast to the reduction of 14-3-3ζ expression, the expression of miR-451, miR-375 and p53 increased in spina bifida rat fetuses.Conclusions/SignificanceThese data suggest that the reduced expression of 14-3-3ζ plays a role in the excessive apoptosis that occurs in spina bifida and may be partly regulated by the over-expression of miR-451 and miR-375, and the consequent up-regulation of p53 might further promote apoptosis in spina bifida.

High-throughput deep screening and identification of four peripheral leucocyte microRNAs as novel potential combination biomarkers for preeclampsia

“…Overexpression of miR-451 could significantly inhibit growth and induce cell apoptosis. 41 , 42 The downregulated miR-451 has been reported in other studies. 43 , 44 In our study, the upregulated miR-451 might be responsible for placenta hypoxia in the PE group.…”

Disruption of melatonin synthesis is associated with impaired 14-3-3 and miR-451 levels in patients with autism spectrum disorders

“…Since 14-3-3ζ is one of the isoforms most implicated in the regulation of melatonin synthesis 23 , 24 , 36 and since 14-3-3ζ was reported to be repressed by miR-451 25 – 27 , we measured this micro-RNA in plasma and pineals. Plasmatic and pineal miR-451 were significantly increased in patients with ASD compared to controls (Fig.…”

“…In humans, it is considered to involve post-translational mechanisms, including AANAT phosphorylation, and stabilization of both AANAT and ASMT by formation of complexes with 14-3-3ζ proteins 23 , 24 . In addition, 14-3-3ζ protein has been shown to be a target of miR-451 25 – 27 which confers resistance to the deleterious effects of oxidative stress, which may play a role in the pathogenesis of ASD, promoting neuronal damage in genetically predisposed individuals 4 .

Figure 1 Exploration of the serotonin-melatonin pathway in the blood.

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Downregulation of microRNA-451 in non-alcoholic steatohepatitis inhibits fatty acid-induced proinflammatory cytokine production through the AMPK/AKT pathway

“…Previous studies have demonstrated that miR-451 targets the 3 UTR mRNAs of Cab39 (Godlewski et al, 2010;Tian et al, 2012), OGT (Bergamaschi and Katzenellenbogen, 2012) and Ywhaz (Wu et al, 2013;Zhang et al, 2012) in multiple cell types. To confirm the expression of these target genes regulated by miR-451, qRT-PCR and Western blotting were performed with murine hepatic tissues.…”