Abstract
Cell-type-specific gene targeting with the Cre/loxP system has become an indispensable technique in experimental neuroscience, particularly for the study of late-born glial cells that make myelin. A plethora of conditional mutants and Cre-expressing mouse lines is now available to the research community that allows laboratories to readily engage in in vivo analyses of oligodendrocytes and their precursor cells. This chapter summarizes concepts and strategies in targeting myelinating glial cells in mice for mutagenesis or imaging, and provides an overview of the most important Cre driver lines successfully used in this rapidly growing field.
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References
Capecchi MR (2005) Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century. Nat Rev Genet 6(6):507–512
Thomas KR, Capecchi MR (1987) Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51(3):503–512
Giese KP et al (1992) Mouse P0 gene disruption leads to hypomyelination, abnormal expression of recognition molecules, and degeneration of myelin and axons. Cell 71(4):565–576
Klugmann M et al (1997) Assembly of CNS myelin in the absence of proteolipid protein. Neuron 18(1):59–70
Lappe-Siefke C et al (2003) Disruption of Cnp1 uncouples oligodendroglial functions in axonal support and myelination. Nat Genet 33(3):366–374
Xin M et al (2005) Myelinogenesis and axonal recognition by oligodendrocytes in brain are uncoupled in Olig1-null mice. J Neurosci 25(6):1354–1365
Emery B et al (2009) Myelin gene regulatory factor is a critical transcriptional regulator required for CNS myelination. Cell 138(1):172–185
Skarnes WC et al (2011) A conditional knockout resource for the genome-wide study of mouse gene function. Nature 474(7351):337–342
Goebbels S et al (2006) Genetic targeting of principal neurons in neocortex and hippocampus of NEX-Cre mice. Genesis 44(12):611–621
Lu QR et al (2002) Common developmental requirement for Olig function indicates a motor neuron/oligodendrocyte connection. Cell 109(1):75–86
Britsch S et al (2001) The transcription factor Sox10 is a key regulator of peripheral glial development. Genes Dev 15(1):66–78
Van de Putte T et al (2003) Mice lacking ZFHX1B, the gene that codes for Smad-interacting protein-1, reveal a role for multiple neural crest cell defects in the etiology of Hirschsprung disease-mental retardation syndrome. Am J Hum Genet 72(2):465–470
Donohoe ME et al (1999) Targeted disruption of mouse Yin Yang 1 transcription factor results in peri-implantation lethality. Mol Cell Biol 19(10):7237–7244
Kotch LE et al (1999) Defective vascularization of HIF-1alpha-null embryos is not associated with VEGF deficiency but with mesenchymal cell death. Dev Biol 209(2):254–267
Dymecki SM (1996) Flp recombinase promotes site-specific DNA recombination in embryonic stem cells and transgenic mice. Proc Natl Acad Sci U S A 93(12):6191–6196
Sauer B, Henderson N (1988) Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc Natl Acad Sci U S A 85(14):5166–5170
Gu H et al (1994) Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting. Science 265(5168):103–106
Goebbels S et al (2010) Elevated phosphatidylinositol 3,4,5-trisphosphate in glia triggers cell-autonomous membrane wrapping and myelination. J Neurosci 30(26):8953–8964
Harrington EP et al (2010) Oligodendrocyte PTEN is required for myelin and axonal integrity, not remyelination. Ann Neurol 68(5):703–716
Wahl SE et al (2014) Mammalian target of rapamycin promotes oligodendrocyte differentiation, initiation and extent of CNS myelination. J Neurosci 34(13):4453–4465
Kang SH et al (2013) Degeneration and impaired regeneration of gray matter oligodendrocytes in amyotrophic lateral sclerosis. Nat Neurosci 16(5):571–579
LoPresti P (2015) Inducible expression of a truncated form of tau in oligodendrocytes elicits gait abnormalities and a decrease in myelin: implications for selective CNS degenerative diseases. Neurochem Res 40(11):2188–2199
Gossen M, Bujard H (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A 89(12):5547–5551
Gossen M et al (1995) Transcriptional activation by tetracyclines in mammalian cells. Science 268(5218):1766–1769
Schonig K, Freundlieb S, Gossen M (2013) Tet-transgenic rodents: a comprehensive, up-to date database. Transgenic Res 22(2):251–254
Goebbels S et al (2005) Cre/loxP-mediated inactivation of the bHLH transcription factor gene NeuroD/BETA2. Genesis 42(4):247–252
Nakagawa Y et al (2016) Ultra-superovulation for the CRISPR-Cas9-mediated production of gene-knockout, single-amino-acid-substituted, and floxed mice. Biol Open 5(8):1142–1148
Quadros RM et al (2017) Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins. Genome Biol 18(1):92
Yang H, Wang H, Jaenisch R (2014) Generating genetically modified mice using CRISPR/Cas-mediated genome engineering. Nat Protoc 9(8):1956–1968
Meyers EN, Lewandoski M, Martin GR (1998) An Fgf8 mutant allelic series generated by Cre- and Flp-mediated recombination. Nat Genet 18(2):136–141
Farley FW et al (2000) Widespread recombinase expression using FLPeR (flipper) mice. Genesis 28(3–4):106–110
Holzenberger M et al (2000) Cre-mediated germline mosaicism: a method allowing rapid generation of several alleles of a target gene. Nucleic Acids Res 28(21):E92
Lakso M et al (1996) Efficient in vivo manipulation of mouse genomic sequences at the zygote stage. Proc Natl Acad Sci U S A 93(12):5860–5865
Umans L et al (2003) Generation of a floxed allele of Smad5 for cre-mediated conditional knockout in the mouse. Genesis 37(1):5–11
Xu X et al (2001) Direct removal in the mouse of a floxed neo gene from a three-loxP conditional knockout allele by two novel approaches. Genesis 30(1):1–6
Ringwald M et al (2011) The IKMC web portal: a central point of entry to data and resources from the International Knockout Mouse Consortium. Nucleic Acids Res 39(Database issue):D849–D855
Rivers LE et al (2008) PDGFRA/NG2 glia generate myelinating oligodendrocytes and piriform projection neurons in adult mice. Nat Neurosci 11(12):1392–1401
Kang SH et al (2010) NG2+ CNS glial progenitors remain committed to the oligodendrocyte lineage in postnatal life and following neurodegeneration. Neuron 68(4):668–681
Zhu X, Bergles DE, Nishiyama A (2008) NG2 cells generate both oligodendrocytes and gray matter astrocytes. Development 135(1):145–157
Zhu X et al (2011) Age-dependent fate and lineage restriction of single NG2 cells. Development 138(4):745–753
Huang W et al (2014) Novel NG2-CreERT2 knock-in mice demonstrate heterogeneous differentiation potential of NG2 glia during development. Glia 62(6):896–913
Battiste J et al (2007) Ascl1 defines sequentially generated lineage-restricted neuronal and oligodendrocyte precursor cells in the spinal cord. Development 134(2):285–293
Niwa-Kawakita M et al (2000) Targeted expression of Cre recombinase to myelinating cells of the central nervous system in transgenic mice. Genesis 26(2):127–129
Hisahara S et al (2000) Targeted expression of baculovirus p35 caspase inhibitor in oligodendrocytes protects mice against autoimmune-mediated demyelination. EMBO J 19(3):341–348
Gow A (2011) Using temporal genetic switches to synchronize the unfolded protein response in cell populations in vivo. Methods Enzymol 491:143–161
Michalski JP et al (2011) The proteolipid protein promoter drives expression outside of the oligodendrocyte lineage during embryonic and early postnatal development. PLoS One 6(5):e19772
Delaunay D et al (2008) Early neuronal and glial fate restriction of embryonic neural stem cells. J Neurosci 28(10):2551–2562
Delaunay D et al (2009) Genetic tracing of subpopulation neurons in the prethalamus of mice (Mus musculus). J Comp Neurol 512(1):74–83
Leone DP et al (2003) Tamoxifen-inducible glia-specific Cre mice for somatic mutagenesis in oligodendrocytes and Schwann cells. Mol Cell Neurosci 22(4):430–440
Doerflinger NH, Macklin WB, Popko B (2003) Inducible site-specific recombination in myelinating cells. Genesis 35(1):63–72
Fruhbeis C et al (2013) Neurotransmitter-triggered transfer of exosomes mediates oligodendrocyte-neuron communication. PLoS Biol 11(7):e1001604
Hovelmeyer N et al (2005) Apoptosis of oligodendrocytes via Fas and TNF-R1 is a key event in the induction of experimental autoimmune encephalomyelitis. J Immunol 175(9):5875–5884
Zou Y et al (2014) Oligodendrocyte precursor cell-intrinsic effect of Rheb1 controls differentiation and mediates mTORC1-dependent myelination in brain. J Neurosci 34(47):15764–15778
Kessaris N et al (2006) Competing waves of oligodendrocytes in the forebrain and postnatal elimination of an embryonic lineage. Nat Neurosci 9(2):173–179
Zawadzka M et al (2010) CNS-resident glial progenitor/stem cells produce Schwann cells as well as oligodendrocytes during repair of CNS demyelination. Cell Stem Cell 6(6):578–590
Matsuoka T et al (2005) Neural crest origins of the neck and shoulder. Nature 436(7049):347–355
McKenzie IA et al (2014) Motor skill learning requires active central myelination. Science 346(6207):318–322
Simon C et al (2012) Sox10-iCreERT2: a mouse line to inducibly trace the neural crest and oligodendrocyte lineage. Genesis 50(6):506–515
Stine ZE et al (2009) Oligodendroglial and pan-neural crest expression of Cre recombinase directed by Sox10 enhancer. Genesis 47(11):765–770
Silbereis JC et al (2014) Olig1 function is required to repress dlx1/2 and interneuron production in Mammalian brain. Neuron 81(3):574–587
Kawaguchi D et al (2016) Generation and analysis of an improved Foxg1-IRES-Cre driver mouse line. Dev Biol 412(1):139–147
Gorski JA et al (2002) Cortical excitatory neurons and glia, but not GABAergic neurons, are produced in the Emx1-expressing lineage. J Neurosci 22(15):6309–6314
Shimshek DR et al (2002) Codon-improved Cre recombinase (iCre) expression in the mouse. Genesis 32(1):19–26
Dumas L et al (2015) Multicolor analysis of oligodendrocyte morphology, interactions, and development with Brainbow. Glia 63(4):699–717
Traka M et al (2016) Oligodendrocyte death results in immune-mediated CNS demyelination. Nat Neurosci 19(1):65–74
Crawford AH et al (2016) Developmental origin of oligodendrocyte lineage cells determines response to demyelination and susceptibility to age-associated functional decline. Cell Rep 15:761–773
Tripathi RB et al (2011) Dorsally and ventrally derived oligodendrocytes have similar electrical properties but myelinate preferred tracts. J Neurosci 31(18):6809–6819
Nave KA, Ehrenreich H (2014) Myelination and oligodendrocyte functions in psychiatric diseases. JAMA Psychiat 71(5):582–584
Brocard J et al (1997) Spatio-temporally controlled site-specific somatic mutagenesis in the mouse. Proc Natl Acad Sci U S A 94(26):14559–14563
Feil R et al (1996) Ligand-activated site-specific recombination in mice. Proc Natl Acad Sci U S A 93(20):10887–10890
Feil R et al (1997) Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding domains. Biochem Biophys Res Commun 237(3):752–757
Forni PE et al (2006) High levels of Cre expression in neuronal progenitors cause defects in brain development leading to microencephaly and hydrocephaly. J Neurosci 26(37):9593–9602
Qiu L, Rivera-Perez JA, Xu Z (2011) A non-specific effect associated with conditional transgene expression based on Cre-loxP strategy in mice. PLoS One 6(5):e18778
Hagemeyer N et al (2012) A myelin gene causative of a catatonia-depression syndrome upon aging. EMBO Mol Med 4(6):528–539
Poggi G et al (2016) Cortical network dysfunction caused by a subtle defect of myelination. Glia 64(11):2025–2040
Akagi K et al (1997) Cre-mediated somatic site-specific recombination in mice. Nucleic Acids Res 25(9):1766–1773
Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21(1):70–71
Lobe CG et al (1999) Z/AP, a double reporter for cre-mediated recombination. Dev Biol 208(2):281–292
De Gasperi R et al (2008) The IRG mouse: a two-color fluorescent reporter for assessing Cre-mediated recombination and imaging complex cellular relationships in situ. Genesis 46(6):308–317
Hartwich H, Satheesh SV, Nothwang HG (2012) A pink mouse reports the switch from red to green fluorescence upon Cre-mediated recombination. BMC Res Notes 5:296
Madisen L et al (2010) A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13(1):133–140
Srinivas S et al (2001) Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol 1:4
Hasegawa Y et al (2013) Novel ROSA26 Cre-reporter knock-in C57BL/6N mice exhibiting green emission before and red emission after Cre-mediated recombination. Exp Anim 62(4):295–304
Clarke LE et al (2012) Properties and fate of oligodendrocyte progenitor cells in the corpus callosum, motor cortex, and piriform cortex of the mouse. J Neurosci 32(24):8173–8185
Dimou L et al (2008) Progeny of Olig2-expressing progenitors in the gray and white matter of the adult mouse cerebral cortex. J Neurosci 28(41):10434–10442
Guo F et al (2010) Pyramidal neurons are generated from oligodendroglial progenitor cells in adult piriform cortex. J Neurosci 30(36):12036–12049
Robins SC et al (2013) Evidence for NG2-glia derived, adult-born functional neurons in the hypothalamus. PLoS One 8(10):e78236
Tsoa RW et al (2014) Spatiotemporally different origins of NG2 progenitors produce cortical interneurons versus glia in the mammalian forebrain. Proc Natl Acad Sci U S A 111(20):7444–7449
Muzumdar MD et al (2007) A global double-fluorescent Cre reporter mouse. Genesis 45(9):593–605
Prigge JR et al (2013) Nuclear double-fluorescent reporter for in vivo and ex vivo analyses of biological transitions in mouse nuclei. Mamm Genome 24:389–399
Rhee JM et al (2006) In vivo imaging and differential localization of lipid-modified GFP-variant fusions in embryonic stem cells and mice. Genesis 44(4):202–218
Aggarwal S et al (2011) A size barrier limits protein diffusion at the cell surface to generate lipid-rich myelin-membrane sheets. Dev Cell 21(3):445–456
Amitai-Lange A et al (2015) A method for lineage tracing of corneal cells using multi-color fluorescent reporter mice. J Vis Exp (106):e53370
Janbandhu VC, Moik D, Fassler R (2014) Cre recombinase induces DNA damage and tetraploidy in the absence of loxP sites. Cell Cycle 13(3):462–470
Genoud S et al (2002) Notch1 control of oligodendrocyte differentiation in the spinal cord. J Cell Biol 158(4):709–718
Tognatta R et al (2017) Transient Cnp expression by early progenitors causes Cre-Lox-based reporter lines to map profoundly different fates. Glia 65(2):342–359
Hirrlinger J et al (2009) Split-CreERT2: temporal control of DNA recombination mediated by split-Cre protein fragment complementation. PLoS One 4(12):e8354
Hirrlinger J et al (2009) Split-cre complementation indicates coincident activity of different genes in vivo. PLoS One 4(1):e4286
Madisen L et al (2012) A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing. Nat Neurosci 15(5):793–802
Gibson EM et al (2014) Neuronal activity promotes oligodendrogenesis and adaptive myelination in the mammalian brain. Science 344(6183):1252304
Micheva KD et al (2016) A large fraction of neocortical myelin ensheathes axons of local inhibitory neurons. Elife 5:e15784
Madisen L et al (2015) Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Neuron 85(5):942–958
Brockschnieder D et al (2006) An improved mouse line for Cre-induced cell ablation due to diphtheria toxin A, expressed from the Rosa26 locus. Genesis 44(7):322–327
Ivanova A et al (2005) In vivo genetic ablation by Cre-mediated expression of diphtheria toxin fragment A. Genesis 43(3):129–135
He M et al (2012) Cell-type-based analysis of microRNA profiles in the mouse brain. Neuron 73(1):35–48
Sanz E et al (2009) Cell-type-specific isolation of ribosome-associated mRNA from complex tissues. Proc Natl Acad Sci U S A 106(33):13939–13944
Pham AH, McCaffery JM, Chan DC (2012) Mouse lines with photo-activatable mitochondria to study mitochondrial dynamics. Genesis 50(11):833–843
Platt RJ et al (2014) CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell 159(2):440–455
Jardi F et al (2017) A shortened tamoxifen induction scheme to induce CreER recombinase without side effects on the male mouse skeleton. Mol Cell Endocrinol 452:57–63
Chen D et al (2002) Tamoxifen and toremifene cause impairment of learning and memory function in mice. Pharmacol Biochem Behav 71(1–2):269–276
Barratt HE et al (2016) Tamoxifen promotes differentiation of oligodendrocyte progenitors in vitro. Neuroscience 319:146–154
Gonzalez GA et al (2016) Tamoxifen accelerates the repair of demyelinated lesions in the central nervous system. Sci Rep 6:31599
Corbo-Rodgers E et al (2012) Oral ivermectin as an unexpected initiator of CreT2-mediated deletion in T cells. Nat Immunol 13(3):197–198
Trevisol et al (2017) Monitoring ATP dynamics in electrically active white matter tracts. eLife 6. pii: e24241
Acknowledgments
We thank Peter Brophy, Brian Popko, Dwight Bergles, Ueli Suter, Bill Richardson, Ori Peles, David Rowitch, and Richard Lu for personal communications on Cre driver lines, members of the Department of Neurogenetics for critical discussion, and Georg Wieser and Ulli Bode for help with the figures. Work in the authors’ laboratories was supported by grants from the DFG (SPP 1757 to S.G. and K.A.N.) and by an European Research Council (ERC) advanced grant (to K.A.N.).
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Goebbels, S., Nave, KA. (2019). Conditional Mutagenesis in Oligodendrocyte Lineage Cells. In: Lyons, D., Kegel, L. (eds) Oligodendrocytes. Methods in Molecular Biology, vol 1936. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9072-6_15
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