Myoblast-derived neuronal cells form glutamatergic neurons in the mouse cerebellum.
Production of neurons from non-neural cells has far-reaching clinical significance. We previously found that myoblasts can be converted to a physiologically active neuronal phenotype by transferring a single recombinant transcription factor, REST-VP16, which directly activates target genes of the transcriptional repressor, REST. However, the neuronal subtype of M-RV cells and whether they can establish synaptic communication in the brain have remained unknown. M-RV cells engineered to express green fluorescent protein (M-RV-GFP) had functional ion channels but did not establish synaptic communication in vitro. However, when transplanted into newborn mice cerebella, a site of extensive postnatal neurogenesis, these cells expressed endogenous cerebellar granule precursors and neuron proteins, such as transient axonal glycoprotein-1, neurofilament, type-III β-tubulin, superior cervical ganglia-clone 10, glutamate receptor-2, and glutamate decarboxylase. Importantly, they exhibited action potentials and were capable of receiving glutamatergic synaptic input, similar to the native cerebellar granule neurons. These results suggest that M-RV-GFP cells differentiate into glutamatergic neurons, an important neuronal subtype, in the postnatal cerebellar milieu. Our findings suggest that although activation of REST-target genes can reprogram myoblasts to assume a general neuronal phenotype, the subtype specificity may then be directed by the brain microenvironment.
Gopalakrishnan
Vidya
V
Department of Pediatrics, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA. vgopalak@mdanderson.org
Bie
Bihua
B
Sinnappah-Kang
Neeta D
ND
Adams
Henry
H
Fuller
Gregory N
GN
Pan
Zhizhong Z
ZZ
Majumder
Sadhan
S
D000818:Animals; D000831:Animals, Newborn; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation; D002531:Cerebellum / Q000166:cytology*; D058969:Contactin 2 / Q000235:genetics / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D032446:Myoblasts / Q000166:cytology*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050777:Stathmin; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D000099229:RE1-Silencing Transcription Factor
D002135:Calcium-Binding Proteins; D058969:Contactin 2; D047908:Intracellular Signaling Peptides and Proteins; D012097:Repressor Proteins; D050777:Stathmin; D014404:Tubulin; D000099229:RE1-Silencing Transcription Factor; C512849:Stmn2 protein, mouse
Curr Stem Cell Res Ther. 2010 Sep;5(3):251-60
20214556
Lancet Neurol. 2009 Sep;8(9):819-29
19679274
Nucleic Acids Res. 2000 Sep 1;28(17):3403-10
10954611
PLoS One. 2009 Sep 24;4(9):e7166
19777066
Philos Trans R Soc Lond B Biol Sci. 2004 May 29;359(1445):851-6
15293812
Stroke. 2007 Feb;38(2 Suppl):810-6
17261745
Mol Cell Biol. 2006 Mar;26(5):1666-78
16478988
Methods Mol Biol. 2008;438:123-34
18369754
Stem Cells. 2009 Oct;27(10):2509-15
19591229
Expert Opin Biol Ther. 2009 Dec;9(12):1487-97
19821796
Genes Dev. 2004 Apr 15;18(8):889-900
15078815
Mol Pain. 2005 Feb 09;1:7
15813995
Nature. 2009 Oct 1;461(7264):649-3
19718018
Curr Biol. 2005 Sep 6;15(17):R665-8
16139198
Philos Trans R Soc Lond B Biol Sci. 2008 Jun 27;363(1500):2079-87
18375377
Eur J Neurosci. 2006 Feb;23(4):945-56
16519659
Curr Opin Neurobiol. 2005 Oct;15(5):500-6
16150588
Science. 1988 Oct 21;242(4877):405-11
3175662
Nat Med. 2006 Nov;12(11):1259-68
17057709
Curr Top Dev Biol. 2005;69:67-99
16243597
Stem Cells Dev. 2006 Jun;15(3):407-21
16846377
Trends Neurosci. 2001 Dec;24(12):680-2
11718852
Nature. 2009 Dec 3;462(7273):587-94
19956253
Nature. 2006 Jun 29;441(7097):1087-93
16810244
Cell. 2005 May 20;121(4):499-501
15907461
Future Neurol. 2009;4(3):317-329
19885378
Development. 1999 Feb;126(3):525-34
9876181
Genes Dev. 2008 Aug 1;22(15):1987-97
18676805
Methods Mol Biol. 2009;549:91-101
19378198
Annu Rev Pharmacol Toxicol. 2004;44:399-421
14744252
Development. 2009 Feb;136(4):509-23
19168672
Exp Neurol. 2000 Sep;165(1):66-76
10964486
Mol Cell Biol. 2004 Sep;24(18):8018-25
15340064
Curr Opin Neurobiol. 2005 Oct;15(5):514-20
16144763
Stem Cells Dev. 2009 May;18(4):591-601
19298173
Cell Stem Cell. 2008 Feb 7;2(2):151-9
18371436
Annu Rev Neurosci. 2009;32:149-84
19555289
J Neurosci. 2005 Apr 13;25(15):3824-32
15829634
Clin Pharmacol Ther. 2007 Oct;82(4):453-60
17713467
Nat Med. 2000 Jul;6(7):826-31
10888935
J Cell Biochem. 2008 Nov 1;105(4):949-55
18668528
Science. 2002 Oct 11;298(5592):425-9
12376704
Prog Neurobiol. 2004 Apr;72(5):295-339
15157725
J Neurosci. 2007 Jun 20;27(25):6729-39
17581960
Curr Protoc Stem Cell Biol. 2009 May;Chapter 2:Unit2D.3
19455542
Neurosurg Focus. 2008;24(3-4):E19
18341395
Cold Spring Harb Symp Quant Biol. 2008;73:227-34
19150961
Curr Opin Cell Biol. 2005 Dec;17(6):664-71
16226449
J Cell Physiol. 2010 Mar;222(3):515-21
20020443
Proc Natl Acad Sci U S A. 2008 Apr 15;105(15):5856-61
18391196
J Neuroimmunol. 2009 Oct 30;215(1-2):73-83
19766326
PLoS One. 2010 Jan 21;5(1):e8814
20098686
Nat Rev Neurosci. 2010 Mar;11(3):176-87
20107441
Annu Rev Cell Dev Biol. 2009;25:253-75
19575663
Cell. 2004 Mar 19;116(6):779-93
15035981
J Intern Med. 2009 Oct;266(4):303-10
19765176
Nat Neurosci. 2008 Aug;11(8):888-93
18587391
Proc Natl Acad Sci U S A. 2004 Jul 13;101(28):10458-63
15240883
Cold Spring Harb Symp Quant Biol. 2008;73:33-8
19022741
Nat Rev Genet. 2007 Jul;8(7):544-54
17572692
Nat Neurosci. 2009 Mar;12(3):311-7
19234456
Nature. 2010 Feb 25;463(7284):1035-41
20107439
Cell Cycle. 2006 Sep;5(17):1929-35
16929174
Cell Stem Cell. 2010 Mar 5;6(3):189-91
20207219