Down-Regulation of Inpp5e Associated With Abnormal Ciliogenesis During Embryonic Neurodevelopment Under Inositol
Huixuan Yue1,2, Shen Li1,2, Jiaxing Qin1
1Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
Insights
Down-regulation of the inositol polyphosphate-5-phosphatase E (Inpp5e) gene impairs primary cilia formation during embryonic neurodevelopment. This is linked to inositol deficiency and neural tube defects (NTDs).
Area of Science:
- Cell Biology
- Developmental Biology
- Genetics
Background:
- The inositol polyphosphate-5-phosphatase E (Inpp5e) enzyme hydrolyzes key phosphoinositides involved in ciliogenesis and embryonic neurodevelopment.
- Mechanisms linking Inpp5e function to these processes, especially under nutrient deficiency, remain largely unknown.
Purpose of the Study:
- To investigate the role of the Inpp5e gene in ciliogenesis during embryonic neurodevelopment using inositol-deficiency models.
- To elucidate the molecular mechanisms underlying inositol deficiency-induced neural tube defects (NTDs).
Main Methods:
- Utilized inositol-deficiency neural tube defects (NTDs) mouse and cell models.
- Employed confocal and scanning electron microscopy to assess primary cilia.
- Analyzed Inpp5e and cilia-related gene expression via immunohistochemistry, western blot, and PCR arrays.
- Quantified inositol and phosphoinositide levels using GC-MS and ELISA.
Main Results:
- Inositol deficiency led to significantly decreased Inpp5e gene expression and reduced PtdIns(3,4)P2 levels in embryonic brain tissues and cell models.
- Observed a reduced number and length of primary cilia in inositol-deficient NIH3T3 cells.
- Down-regulation of key cilia-related genes (Ift80, Mkks, Smo) was noted in inositol-deficient NTDs models, with Smo implicated in NTDs.
Conclusions:
- Down-regulation of Inpp5e is associated with abnormal ciliogenesis during embryonic neurodevelopment under conditions of inositol deficiency.
- Inositol deficiency may disrupt embryonic neurodevelopment through impaired Inpp5e function and subsequent ciliogenesis defects.
- These findings highlight a potential link between Inpp5e, ciliogenesis, and NTDs in the context of inositol metabolism.
Abstract:
The inositol polyphosphate-5-phosphatase E (Inpp5e) gene is located on chromosome 9q34.3. The enzyme it encodes mainly hydrolyzes the 5-phosphate groups of phosphatidylinositol (3,4,5)-trisphosphate (PtdIns (3,4,5) P3) and phosphatidylinositol (4,5)-bisphosphate (PtdIns (4,5)P2), which are closely related to ciliogenesis and embryonic neurodevelopment, through mechanisms that are largely unknown. Here we studied the role of Inpp5e gene in ciliogenesis during embryonic neurodevelopment using inositol-deficiency neural tube defects (NTDs) mouse and cell models. Confocal microscopy and scanning electron microscope were used to examine the number and the length of primary cilia. The dynamic changes of Inpp5e expression in embryonic murine brain tissues were observed during Embryonic Day 10.5-13.5 (E 10.5-13.5). Immunohistochemistry, western blot, polymerase chain reaction (PCR) arrays were applied to detect the expression of Inpp5e and cilia-related genes of the embryonic brain tissues in inositol deficiency NTDs mouse. Real-time quantitative PCR (RT-qPCR) was used to validate the candidate genes in cell models. The levels of inositol and PtdIns(3,4) P2 were measured using gas chromatography-mass spectrometry (GC-MS) and enzyme linked immunosorbent assay (ELISA), respectively. Our results showed that the expression levels of Inpp5e gradually decreased in the forebrain tissues of the control embryos, but no stable trend was observed in the inositol deficiency NTDs embryos. Inpp5e expression in inositol deficiency NTDs embryos was significantly decreased compared with the control tissues. The expression levels of Inpp5e gene and the PtdIns (3,4) P2 levels were also significantly decreased in the inositol deficient cell model. A reduced number and length of primary cilia were observed in NIH3T3 cells when inositol deficient. Three important cilia-related genes (Ift80, Mkks, Smo) were down-regulated significantly in the inositol-deficient NTDs mouse and cell models, and Smo was highly involved in NTDs. In summary, these findings suggested that down-regulation of Inpp5e might be associated with abnormal ciliogenesis during embryonic neurodevelopment, under conditions of inositol deficiency.
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