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In Silico Prediction and Validation of Gfap as an miR-3099 Target in Mouse Brain

Neuroscience Bulletin 2017年第4期33卷 373-382页

作者： Shahidee Zainal Abidin Jia-Wen Leong Marzieh Mahmoudi Norshariza Nordin Syahril Abdullah Pike-See Cheah King-Hwa Ling Neuro Biology and Genetics Group Universiti Putra Malaysia43400 SerdangSelangorMalaysia Genetics and Regenerative Medicine Research Center Universiti Putra Malaysia43400 SerdangSelangorMalaysia Medical Genetics Unit Department of Biomedical SciencesUniversiti Putra Malaysia43400 SerdangSelangorMalaysia Department of Human Anatomy Faculty of Medicine and Health SciencesUniversiti Putra Malaysia43400 SerdangSelangorMalaysia Institute of Biological Sciences Faculty of ScienceUniversity of Malaya50603 Kuala LumpurMalaysia

MicroRNAs are small non-coding RNAs that play crucial roles in the regulation of gene expression and protein synthesis during brain development. MiR-3099 is highly expressed throughout embryogenesis, especially in the developing central nervous system. Moreover, miR-3099 is also expressed at a higher level in differentiating neurons in vitro, suggesting that it is a potential regulator during neuronal cell development. This study aimed to predict the target genes of miR-3099 via in-silico analysis using four independent prediction algorithms（miRDB,miRanda, Target Scan, and DIANA-micro-T-CDS） with emphasis on target genes related to brain development and function. Based on the analysis, a total of 3,174 miR-3099 target genes were predicted. Those predicted by at least three algorithms（324 genes） were subjected to DAVID bioinformatics analysis to understand their overallfunctional themes and representation. The analysis revealed that nearly 70% of the target genes were expressed in the nervous system and a significant proportion were associated with transcriptional regulation and protein ubiquitination mechanisms. Comparison of in situ hybridization（ISH） expression patterns of miR-3099 in both published and in-house-generated ISH sections with the ISH sections of target genes from the Allen Brain Atlas identified 7 target genes（Dnmt3a, Gabpa, Gfap, Itga4,Lxn, Smad7, and Tbx18） having expression patterns complementary to miR-3099 in the developing and adult mouse brain samples. Of these, we validated Gfap as a direct downstream target of miR-3099 using the luciferase reporter gene system. In conclusion, we report the successful prediction and validation of Gfap as an miR-3099 target gene using a combination of bioinformatics resources with enrichment of annotations based on functional ontologies and a spatio-temporal expression dataset.

关键词： Target gene Neurogenesis In silico astrogliogenesis Bioinformatics

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Progenies of NG2 glia: what do we learn from transgenic mouse models ?

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Neural Regeneration Research 2021年第1期16卷 43-48页

作者： Qilin Guo Anja Scheller Wenhui Huang Molecular Physiology Center for Integrative Physiology and Molecular Medicine(CIPMM)University of SaarlandHomburgGermany

In the mammalian central nervous system,nerve-glia antigen 2(NG2)glia are considered the fourth glial population in addition to astrocytes,oligodendrocytes and microglia.The fate of NG2 glia in vivo has been carefully studied in several transgenic mouse models using the Cre/loxP strategy.There is a clear agreement that NG2 glia mainly serve as progenitors for oligodendrocytes and a subpopulation of astrocytes mainly in the ventral forebrain,whereas the existence of a neurogenic potential of NG2 glia is lack of adequate evidence.This mini review summarizes the findings from recent studies regarding the fate of NG2 glia during development.We will highlight the age-and-region-dependent heterogeneity of the NG2 glia differentiation potential.We will also discuss putative reasons for inconsistent findings in various transgenic mouse lines of previous studies.

关键词： astrogliogenesis cell fate Cre/loxP system development differentiation embryonic brain neurogenesis NG2 glia oligodendrocyte lineage oligodendrocyte precursor cells

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Co-culture with microglia promotes neural stem cells differentiation into astrocytes

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Chinese Medical Journal 2011年第20期124卷 3394-3398页

作者： GU Feng WANG Juan FU Li MA Yong-jie Department of Breast PathologyTianjin Medical University Cancer Institute and Hospital Key Laboratory of Breast Cancer Prevention and Therapy of the Ministry of Education and Key Laboratory of Cancer Prevention and Therapy of Tianjin Tianjin 300060 China Department of Oncology Tianjin Medical University Cancer Institute and Hospital Key Laboratory of Breast Cancer Prevention and Therapy of the Ministry of Education and Key Laboratory of Cancer Prevention and Therapy of Tianjin Tianjin 300060 China Department of Gynecology Tianjin Central Gynecology and Obstetrics Hosoital Tianiin 300052 China

Background Neural stem cells （NSCs） are a self-renewing and multipotent population of the central nervous system （CNS）, which are active during development and maintain homeostasis and tissue integrity throughout life. Microglias are an immune cell population resident in the CNS, which have crucial physiological functions in the developing and adult CNS. This study aimed to investigate that whether microglia co-cultured with NSCs could promote astrogliogenesis from NSCs. Methods Microglia and NSCs were co-cultured in 24-well insert plates. NSCs were plated in the bottom of the well and microglia in the insert. Fluorescent staining, Western blotting and RT-PCR were used to determine the effect of microglia on NSCs differentiation. Results Co-culture of microglia and NSCs promoted astrogliogenesis from NSCs. Several key genes, such as Notch 1, Notch 2, Notch 3, Hes 5, and NRSFwere downregulated, while the critical genes Idl and Id2 were upregulated. BMP2 and FGF2 were upregulated. Conclusion Microglias act as a regulator of NSCs astrogliogenesis.

关键词： microglia stem cells astrogliogenesis

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