Mechanism of annexin A6 promoting differentiation and development of neural progenitor cells into oligodendrocytes

您当前的位置：

首页 >

文章列表页 >

Mechanism of annexin A6 promoting differentiation and development of neural progenitor cells into oligodendrocytes

Neurosciences | 更新时间：2026-01-09

- Mechanism of annexin A6 promoting differentiation and development of neural progenitor cells into oligodendrocytes
- Journal of Third Military Medical University Vol. 43, Issue 21, Pages: 2343-2349(2021)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)基础医学院: 组织学与胚胎学教研室，重庆市神经科学重点实验室
  2.400037 重庆，陆军军医大学(第三军医大学)第二附属医院妇产科
  3.400038 重庆，陆军军医大学(第三军医大学)基础医学院：组织学与胚胎学教研室，教学实验中心
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(31471148)
- DOI：10.16016/j.1000-5404.202107102
  CLC： R322.85;R329-33;R329.25
- Received：16 July 2021，
  
  Revised：2021-09-01，
  
  Published：15 November 2021
- 稿件说明：
移动端阅览
Yifan LE, Ruicheng FAN, Yang XU, et al. Mechanism of annexin A6 promoting differentiation and development of neural progenitor cells into oligodendrocytes[J]. Journal of Third Military Medical University, 2021, 43(21): 2343-2349.
DOI：

Yifan LE, Ruicheng FAN, Yang XU, et al. Mechanism of annexin A6 promoting differentiation and development of neural progenitor cells into oligodendrocytes[J]. Journal of Third Military Medical University, 2021, 43(21): 2343-2349. DOI： 10.16016/j.1000-5404.202107102.

摘要

目的

2

探讨膜联蛋白A6(annexin A6，AnxA6)调控神经祖细胞(neural progenitor cells

NPCs)定向少突胶质细胞(oligodendrocytes

OLs)分化的作用机制。

方法

2

构建AnxA6基因敲减(AnxA6敲减组)和过表达(AnxA6过表达组)病毒载体，以空载体作为对照组。运用qRT-PCR、Western blot、免疫荧光染色、活细胞钙成像等技术，对比检测AnxA6敲减与过表达后，体外培养的NPCs胞内Ca

2+

的动态变化差异及其对定向分化转录因子Olig2(定向OLs)、Hes1、Ascl1等表达的影响，评估AnxA6对NPCs分化过程的影响。

结果

2

与对照组相比，AnxA6敲减组未见明显AnxA6蛋白条带(

P

<

0.05)，而AnxA6过表达组阳性蛋白条带光密度值显著升高(

P

<

0.01)；NPCs分化1 d后在静息状态下，AnxA6过表达组细胞可见胞内Ca

2+

自发振荡的频率和振幅均高于对照组和AnxA6敲减组；在NPCs分化3 d的细胞中，可见AnxA6过表达组Olig2 mRNA表达升高，而Hes1、Ascl1 mRNA则表达减少；Olig2

荧光阳性细胞的百分率升高，CNPase荧光信号代表的髓鞘分布增大；同时突起形成相关细胞骨架因子Vimentin、Fyn mRNA的表达显著上调(

P

<

0.05)。

结论

2

过表达AnxA6不仅能增强NPCs分化早期细胞内Ca

2+

的动力变化，还促进OLs定向转录因子的表达，加快髓鞘形成。

Abstract

Objective

2

To explore the mechanism of annexin A6 (AnxA6) regulating neural progenitor cells (NPCs) to differentiate into oligodendrocytes (OLs).

Methods

2

AnxA6 gene knockout and AnxA6 overexpression viral vectors were constructed in freshly isolated and cultured NPCs

and then qPCR

Western blotting

immunofluorescence staining and calcium imaging in living cells were used to detect the changes of AnxA6 knockout and overexpression on the expression of transcription factors Olig2

Hes1 and Ascl1 in NPCs differentiation

in vitro

to observe the dynamic changes of intracellular Ca

2+

and to evaluate the effect of AnxA6 on NPC differentiation and maturation.

Results

2

Compared with the normal control group

there was no obvious positive band of AnxA6 protein in the knockdown group (

P

<

0.05)

but the OD value of the protein band in the overexpression group was significantly increased (

P

<

0.01). Under the resting state of NPCs (differentiation for 1 d)

the frequency and amplitude of spontaneous oscillation of intracellular calcium ions were higher than those in the control group and the knockdown group. In the NPCs cells differentiated for 3 d

the mRNA level of Olig2 was increased and those of Hes1 and Ascl1 were decreased in the AnxA6 overexpression group. The percentage of Olig2 positive cells and the immunofluorescence signal intensity of CNPase were higher than those in the control group and knockdown group

and the up-regulated expression of the myelin formation-related factor Vimentin and Fyn mRNA was also elevated (

P

<

0.05).

Conclusion

2

Over expression of AnxA6 not only enhances the dynamic changes of Ca

2+

in NPCs at the early stage of differentiation

but also promotes the expression of directed transcription factors in OLs and accelerates the formation and myelin sheath.

关键词

Keywords

references

CORNELY R, RENTERO C, ENRICH C, et al . Annexin A6 is an organizer of membrane microdomains to regulate receptor localization and signalling[J ] . Iubmb Life, 2011, 63(11): 1009-1017.

TEBAR F, GELABERT-BALDRICH M, HOQUE M, et al . Annexins and endosomal signaling[J ] . Methods Enzymol, 2014, 535: 55-74. DOI: 10.1016/B978-0-12-397925-4.00004-3.

CHOW A, DAVIS A J, GAWLER D J. Identification of a novel protein complex containing annexin Ⅵ, Fyn, Pyk2, and the p120GAP C2 domain[J]. FEBS Lett, 2000, 469(1): 88-92. DOI: 10.1016/S0014-5793(00)01252-7.

KOESE M, RENTERO C, KOTA B P, et al . Annexin A6 is a scaffold for PKCα to promote EGFR inactivation[J ] . Oncogene, 2013, 32(23): 2858-2872. DOI: 10.1038/onc.2012.303.

ZHOU Y B, XUE S H, CHEN Y Y, et al . Probing Ca 2+ -binding capability of viral proteins with the EF-hand motif by grafting approach[J ] . Methods Mol Biol, 2013, 963: 37-53. DOI: 10.1007/978-1-62703-230-8_3..

CROISSANT C, GOUNOU C, BOUVET F, et al . Annexin-A6 in membrane repair of human skeletal muscle cell: a role in the cap subdomain[J ] . Cells, 2020, 9(7): E1742. DOI: 10.3390/cells9071742.

WU C Y, TANEYHILL L A. Annexin a6 modulates chick cranial neural crest cell emigration[J]. PLoS ONE, 2012, 7(9): e44903. DOI: 10.1371/journal.pone.0044903.

MINASHIMA T, SMALL W, MOSS S E, et al . Intracellular modulation of signaling pathways by annexin A6 regulates terminal differentiation of chondrocytes[J ] . J Biol Chem, 2012, 287(18): 14803-14815. DOI: 10.1074/jbc.M111.297861.

GROCHOWSKI C, RADZIKOWSKA E, MACIEJEWSKI R. Neural stem cell therapy-Brief review[J]. Clin Neurol Neurosurg, 2018, 173: 8-14. DOI: 10.1016/j.clineuro.2018.07.013.

FRAICHARD A, CHASSANDE O, BILBAUT G, et al . In vitro differentiation of embryonic stem cells into glial cells and functional neurons[J ] . J Cell Sci, 1995, 108(Pt 10): 3181-3188..

NESS J K, ROMANKO M J, ROTHSTEIN R P, et al . Perinatal hypoxia-ischemia induces apoptotic and excitotoxic death of periventricular white matter oligodendrocyte progenitors[J ] . Dev Neurosci, 2001, 23(3): 203-208. DOI: 10.1159/000046144.

MA T, LI B C, LE Y F, et al . Demyelination contributes to depression comorbidity in a rat model of chronic epilepsy via dysregulation of Olig2/LINGO-1 and disturbance of calcium homeostasis[J ] . Exp Neurol, 2019, 321: 113034. DOI: 10.1016/j.expneurol.2019.113034..

PENA S A, IYENGAR R, ESHRAGHI R S, et al . Gene therapy for neurological disorders: challenges and recent advancements[J ] . J Drug Target, 2020, 28(2): 111-128. DOI: 10.1080/1061186X.2019.1630415.

HSIEH M C, PENG H Y, HO Y C, et al . Transcription repressor Hes1 contributes to neuropathic pain development by modifying CDK9/RNAPII-dependent spinal mGluR5 transcription[J ] . Int J Mol Sci, 2019, 20(17): E4177. DOI: 10.3390/ijms20174177.

DENG M, XIE P, CHEN Z, et al . Mash-1 modified neural stem cells transplantation promotes neural stem cells differentiation into neurons to further improve locomotor functional recovery in spinal cord injury rats[J ] . Gene, 2021, 781: 145528. DOI: 10.1016/j.gene.2021.145528.

PEKNY M, NILSSON M. Astrocyte activation and reactive gliosis[J]. Glia, 2005, 50(4): 427-434. DOI: 10.1002/glia.20207.

PEKNY M, WILHELMSSON U, TATLISUMAK T, et al . Astrocyte activation and reactive gliosis-A new target in stroke?[J ] . Neurosci Lett, 2019, 689: 45-55.

MONASTYRSKAYA K, BABIYCHUK E B, HOSTETTLER A, et al . Plasma membrane-associated annexin A6 reduces Ca 2+ entry by stabilizing the cortical actin cytoskeleton[J ] . J Biol Chem, 2009, 284(25): 17227-17242. DOI: 10.1074/jbc.M109.004457..

DEMONBREUN A R, FALLON K S, OOSTERBAAN C C, et al . Recombinant annexin A6 promotes membrane repair and protects against muscle injury[J ] . J Clin Invest, 2019, 129(11): 4657-4670. DOI: 10.1172/JCI128840.

PCHITSKAYA E, POPUGAEVA E, BEZPROZVANNY I. Calcium signaling and molecular mechanisms underlying neurodegenerative diseases[J]. Cell Calcium, 2018, 70: 87-94. DOI: 10.1016/j.ceca.2017.06.008.

HOQUE M, ELMAGHRABI Y A, KÖSE M, et al . Annexin A6 improves anti-migratory and anti-invasive properties of tyrosine kinase inhibitors in EGFR overexpressing human squamous epithelial cells[J ] . FEBS J, 2020, 287(14): 2961-2978. DOI: 10.1111/febs.15186.

O'MEARA R W, MICHALSKI J P, ANDERSON C, et al . Integrin-linked kinase regulates process extension in oligodendrocytes via control of actin cytoskeletal dynamics[J ] . J Neurosci, 2013, 33(23): 9781-9793. DOI: 10.1523/JNEUROSCI.5582-12.2013..

MATRONE C, PETRILLO F, NASSO R, et al . Fyn tyrosine kinase as harmonizing factor in neuronal functions and dysfunctions[J ] . Int J Mol Sci, 2020, 21(12): E4444. DOI:10.3390/ijms21124444.

0

Views

0

下载量

0

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Related Articles

Saikosaponin A affects differentiation and function of M1/M2 macrophages by regulating NF-κB and STAT6 signaling pathways

Neurogenin 3 silencing promotes the malignant phenotype of gastric cancer cells

Advances of oligodendrocytes and myelination involving in learning and memory

Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124

Co-culture with umbilical vein endothelial cells promotes differentiation of epicardial progenitor cells into vascular smooth muscle cells

Related Author

SHU Yating

LEI Fan

PENG Meiyu

CUI Zhao

Jianbo ZHANG

Haitao GU

Hailin ZHAO

Xi CHEN

Related Institution

Key Laboratory of Immune Microenvironment and Inflammatory Diseases Research in University of Shandong Province， School of Basic Medical Science， Shandong Second Medical University

Department of Gastrointestinal Surgery

Department of Pathology, the Second Affiliated Hospital of Chongqing Medical University

Department of Physiology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University)

Department of Clinical Laboratory, Zhongda Hospital of Southeast University

⁰