In Vitro Assessment of the Gene Expression of EZH-2 and P300 During Motor Neuron Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells.

IF 1 Q4 NEUROSCIENCES

Basic and Clinical Neuroscience Pub Date : 2022-09-01 DOI:10.32598/bcn.2021.2997.1

Marjaneh Motaghed, Davood Sanooghi, Zohreh Bagher, Faezeh Faghihi, Abolfazl Lotfi, Ali Shahbazi, Mohammad Taghi Jogataei

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Abstract

Introduction: Maintenance of neurogenesis depends on the function of some histone-modifying enzymes; including Enhancer of zeste homolog 2 (EZH2) and histone acetyltransferases (P300). The mechanism of epigenetic regulation and gene expression underlying the transition of human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) into MNs has not been fully clarified.

Methods: Two morphogens; sonic hedgehog (Shh: 100 ng/mL) and retinoic acid (RA: 0.01 mM) were involved in the specification of hUCB-MSCs into MNs after MSC characterization using Flow cytometry. Real time-quantitative PCR and immunocytochemistry were performed to find the expression of the genes at the level of mRNA and protein.

Results: The expression of MN-related markers was confirmed at the level of mRNA and protein by induction of differentiation. The results were confirmed by immunocytochemistry and showed those mean cell percentages of 55.33%±15.885% and 49.67%±13.796% could express Islet-1 and ChAT, respectively. The gene expression level of Islet-1 and ChAT was significantly increased in the first and second week of exposure, respectively. After two weeks, the expression level of P300 and EZH-2 genes increased remarkably. No significant expression of Mnx-1 was detected when compared to the control sample.

Conclusion: MN-related markers, Islet-1 and ChAT, were detected in differentiated cells of hUCB-MSCs, supporting the potency of cord blood cells in the regeneration of MN-related disorders. Assessing these epigenetic regulatory genes at the protein level can be suggested to confirm their functional epigenetic modifying effects during motor neuron differentiation.

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人脐血间充质干细胞运动神经元分化过程中EZH-2和P300基因表达的体外评估。

引言：神经发生的维持取决于一些组蛋白修饰酶的功能；包括皮同源物增强剂2（EZH2）和组蛋白乙酰转移酶（P300）。人脐血间充质干细胞（hUCB-MSCs）向MNs转化的表观遗传学调控和基因表达机制尚未完全阐明。方法：两种形态发生剂；sonic hedgehog（Shh:100ng/mL）和维甲酸（RA:0.01mM）在使用流式细胞术对MSC进行表征后参与将hUCB-MSC鉴定为MNs。采用实时定量PCR和免疫细胞化学方法检测基因在mRNA和蛋白水平的表达。结果：MN相关标志物的表达通过分化诱导在mRNA和蛋白水平得到证实。免疫细胞化学结果证实，平均细胞百分比分别为55.33%±15.885%和49.67%±13.796%，可表达Islet-1和ChAT。Islet-1和ChAT的基因表达水平分别在暴露的第一周和第二周显著增加。两周后，P300和EZH-2基因的表达水平显著升高。与对照样品相比，未检测到Mnx-1的显著表达。结论：在hUCB-MSCs的分化细胞中检测到MN相关标志物Islet-1和ChAT，支持脐血细胞在MN相关疾病再生中的效力。可以建议在蛋白质水平上评估这些表观遗传学调控基因，以证实它们在运动神经元分化过程中的功能表观遗传学修饰作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Basic and Clinical Neuroscience NEUROSCIENCES-

CiteScore

2.60

自引率

0.00%

发文量

审稿时长

4 weeks

期刊介绍： BCN is an international multidisciplinary journal that publishes editorials, original full-length research articles, short communications, reviews, methodological papers, commentaries, perspectives and “news and reports” in the broad fields of developmental, molecular, cellular, system, computational, behavioral, cognitive, and clinical neuroscience. No area in the neural related sciences is excluded from consideration, although priority is given to studies that provide applied insights into the functioning of the nervous system. BCN aims to advance our understanding of organization and function of the nervous system in health and disease, thereby improving the diagnosis and treatment of neural-related disorders. Manuscripts submitted to BCN should describe novel results generated by experiments that were guided by clearly defined aims or hypotheses. BCN aims to provide serious ties in interdisciplinary communication, accessibility to a broad readership inside Iran and the region and also in all other international academic sites, effective peer review process, and independence from all possible non-scientific interests. BCN also tries to empower national, regional and international collaborative networks in the field of neuroscience in Iran, Middle East, Central Asia and North Africa and to be the voice of the Iranian and regional neuroscience community in the world of neuroscientists. In this way, the journal encourages submission of editorials, review papers, commentaries, methodological notes and perspectives that address this scope.