title: Development of cerebellar neurons and glias revealed by in utero electroporation: Golgi-like labeling of cerebellar neurons and glias.
abstract: Cerebellar cortical functions rely on precisely arranged cytoarchitectures composed of several distinct types of neurons and glias. Studies have indicated that cerebellar excitatory and inhibitory neurons have distinct spatial origins, the upper rhombic lip (uRL) and ventricular zone (VZ), respectively, and that different types of neurons have different birthdates. However, the spatiotemporal relationship between uRL/VZ progenitors and their final phenotype remains poorly understood due to technical limitations. To address this issue, we performed in utero electroporation (IUE) of fluorescent protein plasmids using mouse embryos to label uRL/VZ progenitors at specific developmental stages, and observed labeled cells at maturity. To overcome any potential dilution of the plasmids caused by progenitor division, we also utilized constructs that enable permanent labeling of cells. Cerebellar neurons and glias were labeled in a Golgi-like manner enabling ready identification of labeled cells. Five types of cerebellar neurons, namely Purkinje, Golgi, Lugaro and unipolar brush cells, large-diameter deep nuclei (DN) neurons, and DN astrocytes were labeled by conventional plasmids, whereas plasmids that enable permanent labeling additionally labeled stellate, basket, and granule cells as well as three types of glias. IUE allows us to label uRL/VZ progenitors at different developmental stages. We found that the five types of neurons and DN astrocytes were labeled in an IUE stage-dependent manner, while stellate, basket, granule cells and three types of glias were labeled regardless of the IUE stage. Thus, the results indicate the IUE is an efficient method to track the development of cerebellar cells from uRL/VZ progenitors facing the ventricular lumen. They also indicate that while the generation of the five types of neurons by uRL/VZ progenitors is regulated in a time-dependent manner, the progenitor pool retains multipotency throughout embryonic development.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kita
authors.forename: Koichi
authors.initials: K
authors.lastname: Kawakami
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Fujio
authors.initials: F
authors.lastname: Murakami
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology; D002525:Cerebellar Cortex / Q000166:cytology* / Q000196:embryology; D018274:Electroporation; D004622:Embryo, Mammalian; D005260:Female; D018014:Gene Transfer Techniques; D006056:Golgi Apparatus / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D058953:Neural Stem Cells / Q000166:cytology*; D009457:Neuroglia / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D011247:Pregnancy
references.citation: Dev Biol. 2008 Oct 15;322(2):345-54
references.pmid: 18723012
references.citation: J Neurosci. 2006 Sep 6;26(36):9184-95
references.pmid: 16957075
references.citation: J Comp Neurol. 1994 Jan 8;339(2):174-80
references.pmid: 8300904
references.citation: Dev Biol. 2009 Apr 15;328(2):422-33
references.pmid: 19217896
references.citation: Curr Opin Neurobiol. 2005 Feb;15(1):29-33
references.pmid: 15721741
references.citation: Genome Biol. 2007;8 Suppl 1:S7
references.pmid: 18047699
references.citation: Neuron. 1996 Jan;16(1):47-54
references.pmid: 8562089
references.citation: Dev Biol. 2007 May 15;305(2):616-24
references.pmid: 17362912
references.citation: Exp Neurol. 1961 Oct;4:277-96
references.pmid: 14473282
references.citation: J Biol Chem. 2010 Jan 1;285(1):373-80
references.pmid: 19887377
references.citation: Brain Res. 1975 Sep 23;95(2-3):503-18
references.pmid: 1156884
references.citation: J Comp Neurol. 1978 May 1;179(1):23-48
references.pmid: 8980716
references.citation: J Comp Neurol. 1960 Apr;114:137-59
references.pmid: 13840582
references.citation: Glia. 2011 Dec;59(12):1946-57
references.pmid: 21901755
references.citation: Dev Cell. 2004 Jul;7(1):133-44
references.pmid: 15239961
references.citation: Nat Neurosci. 2010 Jan;13(1):133-40
references.pmid: 20023653
references.citation: J Comp Neurol. 1980 Oct 1;193(3):815-40
references.pmid: 7002963
references.citation: J Comp Neurol. 2003 Dec 1;467(1):60-79
references.pmid: 14574680
references.citation: J Neurosci. 2011 Jul 27;31(30):11055-69
references.pmid: 21795554
references.citation: Gene. 1991 Dec 15;108(2):193-9
references.pmid: 1660837
references.citation: Neuron. 2007 May 3;54(3):357-69
references.pmid: 17481390
references.citation: Histochem Cell Biol. 2008 Oct;130(4):601-15
references.pmid: 18677503
references.citation: Neuron. 2005 Oct 6;48(1):31-43
references.pmid: 16202707
references.citation: Neuron. 2005 Jan 6;45(1):27-40
references.pmid: 15629700
references.citation: Cell Mol Life Sci. 2003 Jun;60(6):1084-98
references.pmid: 12861377
references.citation: Neuron. 2005 Jul 21;47(2):201-13
references.pmid: 16039563
references.citation: J Comp Neurol. 1966 Oct;128(2):191-208
references.pmid: 5970298
references.citation: Nat Rev Neurosci. 2001 Apr;2(4):287-93
references.pmid: 11283751
references.citation: Development. 2009 Jun;136(12):2049-58
references.pmid: 19439493
references.citation: Neuron. 2005 Oct 6;48(1):17-24
references.pmid: 16202705
references.citation: Anat Embryol (Berl). 1981;162(1):51-67
references.pmid: 7283173
references.citation: Neuroscience. 2004;127(4):845-58
references.pmid: 15312897
references.citation: J Comp Neurol. 2003 May 5;459(3):278-89
references.pmid: 12655510
references.citation: J Neurosci. 2009 May 27;29(21):7079-91
references.pmid: 19474334
references.citation: Trends Neurosci. 2010 Apr;33(4):163-72
references.pmid: 20138673
references.citation: J Comp Neurol. 1992 Jan 1;315(1):74-84
references.pmid: 1371781
references.citation: Mol Cell Neurosci. 2012 Feb;49(2):217-29
references.pmid: 22173107
references.citation: Proc Natl Acad Sci U S A. 2010 Jun 8;107(23):10725-30
references.pmid: 20498066
references.citation: Brain Res. 1971 Dec 10;35(1):17-36
references.pmid: 4109144
references.citation: Dev Growth Differ. 2008 Aug;50(6):499-506
references.pmid: 18482402
references.citation: Neuroscientist. 2008 Feb;14(1):91-100
references.pmid: 17911211
references.citation: Genes Cells. 2010 May;15(5):501-12
references.pmid: 20384787
references.citation: J Comp Neurol. 1996 Jul 08;370(4):536-50
references.pmid: 8807453
references.citation: J Neurocytol. 1999 Feb;28(2):99-123
references.pmid: 10590511
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):5193-8
references.pmid: 17360405
references.citation: Prog Neurobiol. 2004 Apr;72(5):295-339
references.pmid: 15157725
references.citation: Mol Reprod Dev. 1997 Feb;46(2):109-13
references.pmid: 9021742
references.citation: Gene. 2004 Aug 18;338(1):93-8
references.pmid: 15302410
references.citation: J Neurosci. 2009 Feb 4;29(5):1300-11
references.pmid: 19193877
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 29;106(52):22462-7
references.pmid: 20080794
references.citation: J Comp Neurol. 2002 Dec 2;454(1):1-14
references.pmid: 12410614
references.citation: J Neurosci. 2006 Nov 8;26(45):11682-94
references.pmid: 17093090
references.citation: Neurosci Res. 2004 Sep;50(1):13-22
references.pmid: 15288494
references.citation: J Comp Neurol. 1972 Jul;145(3):353-97
references.pmid: 4113154
references.citation: Prog Neurobiol. 2006 Feb-Apr;78(3-5):272-303
references.pmid: 16759785
references.citation: Nucleic Acids Res. 1995 Oct 11;23(19):3816-21
references.pmid: 7479022