EXPRESSION OF EXOGENOUS LIN28 CONTRIBUTES TO PROLIFERATION AND SURVIVAL OF MOUSE PRIMARY CORTICAL NEURONS IN VITRO

被引:21
作者
Bhuiyan, M. I. H. [1 ]
Lee, J. -H. [2 ]
Kim, S. Y. [1 ]
Cho, K. -O. [1 ]
机构
[1] Catholic Univ Korea, Dept Pharmacol, Cell Death Dis Res Ctr, Coll Med, Seoul 137701, South Korea
[2] Catholic Univ Korea, Dept Biochem, Coll Med, Seoul 137701, South Korea
基金
新加坡国家研究基金会;
关键词
LIN28; prosurvival effect; cell proliferation; in utero electroporation; primary cortical neuron; apoptosis; CELL-DEATH; SERUM-FREE; SUBSTANTIA-NIGRA; LET-7; MICRORNA; LIN-28; HOMOLOG; HIPPOCAMPAL; GROWTH; DENSITY; VIVO; BIOGENESIS;
D O I
10.1016/j.neuroscience.2013.06.023
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
LIN28, an RNA-binding protein, is known to be involved in the regulation of many cellular processes, such as embryonic stem cell proliferation, cell fate succession, developmental timing, and oncogenesis. In this study, we investigated the effect of constitutively expressing exogenous LIN28 on neuronal cell proliferation and viability in vitro. Plasmids containing LIN28-green fluorescent protein (GFP) or GFP were introduced into the embryonic mouse brains at E14.5 by in utero electroporation. Two days after electroporation, embryonic cortices were harvested and cultured. It was found that transfected cells stably overexpressed LIN28 in vitro. Viability curve from live cell imaging showed that the number of GFP-expressing cells decreased over time in line with naive primary cortical neurons. In contrast, the number of LIN28-GFP-overexpressing neurons initially increased and remained high at later time-points in culture than GFP-expressing cells. Double immunofluorescence showed that at an early time in culture, the number of Ki-67/GFP double-positive cells was higher in the LIN28-GFP group than that of controls. Moreover, there were significantly lower numbers of condensed nuclei/GFP- and cleaved caspase-3/GFP-positive cells in the LIN28-GFP groups compared to control GFP. Furthermore, it was confirmed that the LIN28-GFP-expressing cells at days in vitro (DIV)13 were neuronal nuclei (NeuN)-positive mature neurons. Finally, the expression of insulin-like growth factor 2 (IGF-2) was induced in LIN28-expressing primary cortical neurons, which was not detected in controls. Taken together, our results indicate that the expression of exogenous LIN28 can promote the proliferation of neural progenitor cells and exert prosurvival effect on primary cortical neurons by inhibiting caspase-dependent apoptosis, possibly via upregulation of IGF-2. (C) 2013 IBRO. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:448 / 458
页数:11
相关论文
共 54 条
[1]   A hippocampal insulin-growth factor 2 pathway regulates the extinction of fear memories [J].
Agis-Balboa, Roberto Carlos ;
Arcos-Diaz, Dario ;
Wittnam, Jessica ;
Govindarajan, Nambirajan ;
Blom, Kim ;
Burkhardt, Susanne ;
Haladyniak, Ulla ;
Agbemenyah, Hope Yao ;
Zovoilis, Athanasios ;
Salinas-Riester, Gabriella ;
Opitz, Lennart ;
Sananbenesi, Farahnaz ;
Fischer, Andre .
EMBO JOURNAL, 2011, 30 (19) :4071-4083
[2]   LIN28 alters cell fate succession and acts independently of the let-7 microRNA during neurogliogenesis in vitro [J].
Balzer, Erica ;
Heine, Christian ;
Jiang, Qiang ;
Lee, Vivian M. ;
Moss, Eric G. .
DEVELOPMENT, 2010, 137 (06) :891-900
[3]   Anthocyanins Extracted from Black Soybean Seed Coat Protect Primary Cortical Neurons against in Vitro Ischemia [J].
Bhuiyan, Mohammad Iqbal Hossain ;
Kim, Joo Youn ;
Ha, Tae Joung ;
Kim, Seong Yun ;
Cho, Kyung-Ok .
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 2012, 35 (07) :999-1008
[4]   The Neuroprotective Potential of Cyanidin-3-glucoside Fraction Extracted from Mulberry Following Oxygen-glucose Deprivation [J].
Bhuiyan, Mohammad Iqbal Hossain ;
Kim, Hyun-Bok ;
Kim, Seong Yun ;
Cho, Kyung-Ok .
KOREAN JOURNAL OF PHYSIOLOGY & PHARMACOLOGY, 2011, 15 (06) :353-361
[5]   SERUM-FREE B27/NEUROBASAL MEDIUM SUPPORTS DIFFERENTIATED GROWTH OF NEURONS FROM THE STRIATUM, SUBSTANTIA-NIGRA, SEPTUM, CEREBRAL-CORTEX, CEREBELLUM, AND DENTATE GYRUS [J].
BREWER, GJ .
JOURNAL OF NEUROSCIENCE RESEARCH, 1995, 42 (05) :674-683
[6]  
Brewer GregoryJ., 1994, Focus, V16, P6
[7]   EGF ENHANCES THE SURVIVAL OF DOPAMINE NEURONS IN RAT EMBRYONIC MESENCEPHALON PRIMARY-CELL CULTURE [J].
CASPER, D ;
MYTILINEOU, C ;
BLUM, M .
JOURNAL OF NEUROSCIENCE RESEARCH, 1991, 30 (02) :372-381
[8]  
Cianfarani Stefano, 2012, Front Endocrinol (Lausanne), V3, P118, DOI 10.3389/fendo.2012.00118
[9]   Genetic Analysis of the Role of the Reprogramming Gene LIN-28 in Human Embryonic Stem Cells [J].
Darr, Henia ;
Benvenisty, Nissim .
STEM CELLS, 2009, 27 (02) :352-362
[10]   Insulin-like growth factor I protects and rescues hippocampal neurons against beta-amyloid- and human amylin-induced toxicity [J].
Dore, S ;
Kar, S ;
Quirion, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (09) :4772-4777