[en] The mature cerebellum controls motor skill precision and participates in other sophisticated brain functions that include learning, cognition, and speech. Different types of GABAergic and glutamatergic cerebellar neurons originate in temporal order from two progenitor niches, the ventricular zone and rhombic lip, which express the transcription factors Ptf1a and Atoh1, respectively. However, the molecular machinery required to specify the distinct neuronal types emanating from these progenitor zones is still unclear. Here, we uncover the transcription factor Olig3 as a major determinant in generating the earliest neuronal derivatives emanating from both progenitor zones in mice. In the rhombic lip, Olig3 regulates progenitor cell proliferation. In the ventricular zone, Olig3 safeguards Purkinje cell specification by curtailing the expression of Pax2, a transcription factor that suppresses the Purkinje cell differentiation program. Our work thus defines Olig3 as a key factor in early cerebellar development.
Lowenstein, Elijah D ; Max-Delbrück-Centrum in the Helmholtz Association, Berlin, Germany.
Rusanova, Aleksandra; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin, ; Institute of Neuroscience, Lobachevsky University of Nizhny Novgorod, Nizhny
Stelzer, Jonas; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin,
Hernaiz-Llorens, Marc ; Max-Delbrück-Centrum in the Helmholtz Association, Berlin, Germany.
Schroer, Adrian E; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin,
Epifanova, Ekaterina ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques ; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin, ; Institute of Neuroscience, Lobachevsky University of Nizhny Novgorod, Nizhny
Bladt, Francesca; Max-Delbrück-Centrum in the Helmholtz Association, Berlin, Germany.
Isik, Eser Göksu; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin,
Buchert, Sven; Max-Delbrück-Centrum in the Helmholtz Association, Berlin, Germany.
Jia, Shiqi; Max-Delbrück-Centrum in the Helmholtz Association, Berlin, Germany. ; The First Affiliated Hospital of Jinan University, Guangzhou province, Guangzhou,
Tarabykin, Victor; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin, ; Institute of Neuroscience, Lobachevsky University of Nizhny Novgorod, Nizhny
Hernandez-Miranda, Luis R ; Max-Delbrück-Centrum in the Helmholtz Association, Berlin, Germany. ; Institute for Cell Biology and Neurobiology, Charité Universitätsmedizin Berlin
Alder J, Cho NK, Hatten ME. 1996. Embryonic precursor cells from the rhombic lip are specified to a cerebellar granule neuron identity. Neuron 17:389–399. DOI: https://doi.org/10.1016/S0896-6273(00)80172-5, PMID: 8816703
Atchley WR, Fitch WM. 1997. A natural classification of the basic helix-loop-helix class of transcription factors. PNAS 94:5172–5176. DOI: https://doi.org/10.1073/pnas.94.10.5172, PMID: 9144210
Baker NE, Brown NL. 2018. All in the family: proneural bHLH genes and neuronal diversity. Development 145: dev159426. DOI: https://doi.org/10.1242/dev.159426, PMID: 29720483
Ben-Arie N, Bellen HJ, Armstrong DL, McCall AE, Gordadze PR, Guo Q, Matzuk MM, Zoghbi HY. 1997. Math1 is essential for genesis of cerebellar granule neurons. Nature 390:169–172. DOI: https://doi.org/10.1038/36579, PMID: 9367153
Ben-Arie N, Hassan BA, Bermingham NA, Malicki DM, Armstrong D, Matzuk M, Bellen HJ, Zoghbi HY. 2000. Functional conservation of atonal and Math1 in the CNS and PNS. Development 127:1039–1048. PMID: 10662643
Bertrand N, Castro DS, Guillemot F. 2002. Proneural genes and the specification of neural cell types. Nature Reviews Neuroscience 3:517–530. DOI: https://doi.org/10.1038/nrn874, PMID: 12094208
Butts T, Green MJ, Wingate RJ. 2014. Development of the cerebellum: simple steps to make a ’little brain’. Development 141:4031–4041. DOI: https://doi.org/10.1242/dev.106559, PMID: 25336734
Chizhikov VV, Lindgren AG, Currle DS, Rose MF, Monuki ES, Millen KJ. 2006. The roof plate regulates cerebellar cell-type specification and proliferation. Development 133:2793–2804. DOI: https://doi.org/10.1242/dev. 02441, PMID: 16790481
Comai G, Tajbakhsh S. 2014. Molecular and cellular regulation of skeletal myogenesis. Current Topics in Developmental Biology 110:1–73. DOI: https://doi.org/10.1016/B978-0-12-405943-6.00001-4, PMID: 25248473
Dahmane N, Ruiz i Altaba A. 1999. Sonic hedgehog regulates the growth and patterning of the cerebellum. Development 126:3089–3100. PMID: 10375501
Dennis DJ, Han S, Schuurmans C. 2019. bHLH transcription factors in neural development, disease, and reprogramming. Brain Research 1705:48–65. DOI: https://doi.org/10.1016/j.brainres.2018.03.013, PMID: 2 9544733
Dessaud E, Yang LL, Hill K, Cox B, Ulloa F, Ribeiro A, Mynett A, Novitch BG, Briscoe J. 2007. Interpretation of the sonic hedgehog morphogen gradient by a temporal adaptation mechanism. Nature 450:717–720. DOI: https://doi.org/10.1038/nature06347
Dokucu ME, Zipursky SL, Cagan RL. 1996. Atonal, rough and the resolution of proneural clusters in the developing Drosophila retina. Development 122:4139–4147. PMID: 9012533
Englund C, Kowalczyk T, Daza RA, Dagan A, Lau C, Rose MF, Hevner RF. 2006. Unipolar brush cells of the cerebellum are produced in the rhombic lip and migrate through developing white matter. Journal of Neuroscience 26:9184–9195. DOI: https://doi.org/10.1523/JNEUROSCI.1610-06.2006, PMID: 16957075
Fink AJ. 2006. Development of the deep cerebellar nuclei: transcription factors and cell migration from the rhombic lip. Journal of Neuroscience 26:3066–3076. DOI: https://doi.org/10.1523/JNEUROSCI.5203-05.2006
Gazit R, Krizhanovsky V, Ben-Arie N. 2004. Math1 controls cerebellar granule cell differentiation by regulating multiple components of the notch signaling pathway. Development 131:903–913. DOI: https://doi.org/10. 1242/dev.00982, PMID: 14757642
Grimaldi P, Parras C, Guillemot F, Rossi F, Wassef M. 2009. Origins and control of the differentiation of inhibitory interneurons and Glia in the cerebellum. Developmental Biology 328:422–433. DOI: https://doi.org/10.1016/j. ydbio.2009.02.008, PMID: 19217896
Hallonet ME, Teillet MA, Le Douarin NM. 1990. A new approach to the development of the cerebellum provided by the quail-chick marker system. Development 108:19–31.
Hashimoto M, Mikoshiba K. 2003. Mediolateral compartmentalization of the cerebellum is determined on the "birth date" of Purkinje cells. The Journal of Neuroscience 23:11342–11351. DOI: https://doi.org/10.1523/JNEUROSCI.23-36-11342.2003, PMID: 14672998
Hernández-Miranda LR, Parnavelas JG, Chiara F. 2010. Molecules and mechanisms involved in the generation and migration of cortical interneurons. ASN Neuro 2::AN20090053. DOI: https://doi.org/10.1042/AN20090053
Hernández-Miranda LR, Cariboni A, Faux C, Ruhrberg C, Cho JH, Cloutier JF, Eickholt BJ, Parnavelas JG, Andrews WD. 2011. Robo1 regulates semaphorin signaling to guide the migration of cortical interneurons through the ventral forebrain. Journal of Neuroscience 31:6174–6187. DOI: https://doi.org/10.1523/JNEUROSCI.5464-10.2011, PMID: 21508241
Hernandez-Miranda LR, Müller T, Birchmeier C. 2017a. The dorsal spinal cord and hindbrain: from developmental mechanisms to functional circuits. Developmental Biology 432:34–42. DOI: https://doi.org/10. 1016/j.ydbio.2016.10.008, PMID: 27742210
Hernandez-Miranda LR, Ruffault PL, Bouvier JC, Murray AJ, Morin-Surun MP, Zampieri N, Cholewa-Waclaw JB, Ey E, Brunet JF, Champagnat J, Fortin G, Birchmeier C. 2017b. Genetic identification of a hindbrain nucleus essential for innate vocalization. PNAS 114:8095–8100. DOI: https://doi.org/10.1073/pnas.1702893114, PMID: 28698373
Hippenmeyer S, Vrieseling E, Sigrist M, Portmann T, Laengle C, Ladle DR, Arber S. 2005. A developmental switch in the response of DRG neurons to ETS transcription factor signaling. PLOS Biology 3:e159. DOI: https://doi.org/10.1371/journal.pbio.0030159, PMID: 15836427
Hoshino M, Nakamura S, Mori K, Kawauchi T, Terao M, Nishimura YV, Fukuda A, Fuse T, Matsuo N, Sone M, Watanabe M, Bito H, Terashima T, Wright CV, Kawaguchi Y, Nakao K, Nabeshima Y. 2005. Ptf1a, a bHLH transcriptional gene, defines GABAergic neuronal fates in cerebellum. Neuron 47:201–213. DOI: https://doi. org/10.1016/j.neuron.2005.06.007, PMID: 16039563
Imayoshi I, Kageyama R. 2014. bHLH factors in self-renewal, Multipotency, and fate choice of neural progenitor cells. Neuron 82:9–23. DOI: https://doi.org/10.1016/j.neuron.2014.03.018, PMID: 24698265
Jensen P. 2004. Analysis of cerebellar development in math1 null embryos and chimeras. Journal of Neuroscience 24:2202–2211. DOI: https://doi.org/10.1523/JNEUROSCI.3427-03.2004
Jessell TM. 2000. Neuronal specification in the spinal cord: inductive signals and transcriptional codes. Nature Reviews Genetics 1:20–29. DOI: https://doi.org/10.1038/35049541, PMID: 11262869
Jones S. 2004. An overview of the basic helix-loop-helix proteins. Genome Biology 5:226. DOI: https://doi.org/10.1186/gb-2004-5-6-226, PMID: 15186484
Ju J, Liu Q, Zhang Y, Liu Y, Jiang M, Zhang L, He X, Peng C, Zheng T, Lu QR, Li H. 2016. Olig2 regulates purkinje cell generation in the early developing mouse cerebellum. Scientific Reports 6:30711. DOI: https://doi.org/10. 1038/srep30711, PMID: 27469598
Leto K, Carletti B, Williams IM, Magrassi L, Rossi F. 2006. Different types of cerebellar GABAergic interneurons originate from a common pool of multipotent progenitor cells. Journal of Neuroscience 26:11682–11694. DOI: https://doi.org/10.1523/JNEUROSCI.3656-06.2006, PMID: 17093090
Liu Z, Li H, Hu X, Yu L, Liu H, Han R, Colella R, Mower GD, Chen Y, Qiu M. 2008. Control of precerebellar neuron development by Olig3 bHLH transcription factor. Journal of Neuroscience 28:10124–10133. DOI: https://doi. org/10.1523/JNEUROSCI.3769-08.2008, PMID: 18829970
Liu ZR, Shi M, Hu ZL, Zheng MH, Du F, Zhao G, Ding YQ. 2010. A refined map of early gene expression in the dorsal rhombomere 1 of mouse embryos. Brain Research Bulletin 82:74–82. DOI: https://doi.org/10.1016/j. brainresbull.2010.02.010, PMID: 20206242
Lizen B, Claus M, Jeannotte L, Rijli FM, Gofflot F. 2015. Perinatal induction of cre recombination with tamoxifen. Transgenic Research 24:1065–1077. DOI: https://doi.org/10.1007/s11248-015-9905-5, PMID: 26395370
Machold R, Klein C, Fishell G. 2011. Genes expressed in Atoh1 neuronal lineages arising from the r1/isthmus rhombic lip. Gene Expression Patterns 11:349–359. DOI: https://doi.org/10.1016/j.gep.2011.03.007, PMID: 21440680
Machold R, Fishell G. 2005. Math1 is expressed in temporally discrete pools of cerebellar rhombic-lip neural progenitors. Neuron 48:17–24. DOI: https://doi.org/10.1016/j.neuron.2005.08.028, PMID: 16202705
Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, Ng LL, Palmiter RD, Hawrylycz MJ, Jones AR, Lein ES, Zeng H. 2010. A robust and high-throughput cre reporting and characterization system for the whole mouse brain. Nature Neuroscience 13:133–140. DOI: https://doi.org/10.1038/nn.2467, PMID: 20023653
Maricich SM, Herrup K. 1999. Pax-2 expression defines a subset of GABAergic interneurons and their precursors in the developing murine cerebellum. Journal of Neurobiology 41:281–294. DOI: https://doi.org/10.1002/(SICI) 1097-4695(19991105)41:2<281::AID-NEU10>3.0.CO;2-5, PMID: 10512984
Mattar P, Langevin LM, Markham K, Klenin N, Shivji S, Zinyk D, Schuurmans C. 2008. Basic helix-loop-helix transcription factors cooperate to specify a cortical projection neuron identity. Molecular and Cellular Biology 28:1456–1469. DOI: https://doi.org/10.1128/MCB.01510-07, PMID: 18160702
Millen KJ, Steshina EY, Iskusnykh IY, Chizhikov VV. 2014. Transformation of the cerebellum into more ventral brainstem fates causes cerebellar agenesis in the absence of Ptf1a function. PNAS 111:E1777–E1786. DOI: https://doi.org/10.1073/pnas.1315024111, PMID: 24733890
Millet S, Bloch-Gallego E, Simeone A, Alvarado-Mallart RM. 1996. The caudal limit of Otx2 gene expression as a marker of the midbrain/hindbrain boundary: a study using in situ hybridisation and chick/quail homotopic grafts. Development 122:3785–3797. PMID: 9012500
Morales D, Hatten ME. 2006. Molecular markers of neuronal progenitors in the embryonic cerebellar anlage. Journal of Neuroscience 26:12226–12236. DOI: https://doi.org/10.1523/JNEUROSCI.3493-06.2006, PMID: 17122047
Müller T, Anlag K, Wildner H, Britsch S, Treier M, Birchmeier C. 2005. The bHLH factor Olig3 coordinates the specification of dorsal neurons in the spinal cord. Genes & Development 19:733–743. DOI: https://doi.org/10. 1101/gad.326105, PMID: 15769945
Ross SE, Greenberg ME, Stiles CD. 2003. Basic helix-loop-helix factors in cortical development. Neuron 39:13– 25. DOI: https://doi.org/10.1016/S0896-6273(03)00365-9, PMID: 12848929
Saito T, Nakatsuji N. 2001. Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Developmental Biology 240:237–246. DOI: https://doi.org/10.1006/dbio.2001.0439, PMID: 11784059
Sekerková G, Ilijic E, Mugnaini E. 2004. Time of origin of unipolar brush cells in the rat cerebellum as observed by prenatal bromodeoxyuridine labeling. Neuroscience 127:845–858. DOI: https://doi.org/10.1016/j. neuroscience.2004.05.050, PMID: 15312897
Sellick GS, Barker KT, Stolte-Dijkstra I, Fleischmann C, Coleman RJ, Garrett C, Gloyn AL, Edghill EL, Hattersley AT, Wellauer PK, Goodwin G, Houlston RS. 2004. Mutations in PTF1A cause pancreatic and cerebellar agenesis. Nature Genetics 36:1301–1305. DOI: https://doi.org/10.1038/ng1475, PMID: 15543146
Seto Y, Nakatani T, Masuyama N, Taya S, Kumai M, Minaki Y, Hamaguchi A, Inoue YU, Inoue T, Miyashita S, Fujiyama T, Yamada M, Chapman H, Campbell K, Magnuson MA, Wright CV, Kawaguchi Y, Ikenaka K, Takebayashi H, Ishiwata S, et al. 2014. Temporal identity transition from purkinje cell progenitors to GABAergic interneuron progenitors in the cerebellum. Nature Communications 5:3337. DOI: https://doi.org/10.1038/ncomms4337, PMID: 24535035
Shiraishi A, Muguruma K, Sasai Y. 2017. Generation of thalamic neurons from mouse embryonic stem cells. Development 144:1211–1220. DOI: https://doi.org/10.1242/dev.144071, PMID: 28219951
Skinner MK, Rawls A, Wilson-Rawls J, Roalson EH. 2010. Basic helix-loop-helix transcription factor gene family phylogenetics and nomenclature. Differentiation 80:1–8. DOI: https://doi.org/10.1016/j.diff.2010.02.003, PMID: 20219281
Sommer L, Ma Q, Anderson DJ. 1996. Neurogenins, a novel family of atonal-related bHLH transcription factors, are putative mammalian neuronal determination genes that reveal progenitor cell heterogeneity in the developing CNS and PNS. Molecular and Cellular Neuroscience 8:221–241. DOI: https://doi.org/10.1006/mcne.1996.0060, PMID: 9000438
Stevens JD, Roalson EH, Skinner MK. 2008. Phylogenetic and expression analysis of the basic helix-loop-helix transcription factor gene family: genomic approach to cellular differentiation. Differentiation 76:1006–1042. DOI: https://doi.org/10.1111/j.1432-0436.2008.00285.x, PMID: 18557763
Storm R, Cholewa-Waclaw J, Reuter K, Bröhl D, Sieber M, Treier M, Müller T, Birchmeier C. 2009. The bHLH transcription factor Olig3 marks the dorsal neuroepithelium of the hindbrain and is essential for the development of brainstem nuclei. Development 136:295–305. DOI: https://doi.org/10.1242/dev.027193, PMID: 19088088
Sudarov A, Turnbull RK, Kim EJ, Lebel-Potter M, Guillemot F, Joyner AL. 2011. Ascl1 genetics reveals insights into cerebellum local circuit assembly. Journal of Neuroscience 31:11055–11069. DOI: https://doi.org/10.1523/JNEUROSCI.0479-11.2011, PMID: 21795554
Susaki EA, Tainaka K, Perrin D, Yukinaga H, Kuno A, Ueda HR. 2015. Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging. Nature Protocols 10:1709–1727. DOI: https://doi.org/10.1038/nprot. 2015.085, PMID: 26448360
Takebayashi H, Yoshida S, Sugimori M, Kosako H, Kominami R, Nakafuku M, Nabeshima Y. 2000. Dynamic expression of basic helix-loop-helix olig family members: implication of Olig2 in neuron and oligodendrocyte differentiation and identification of a new member, Olig3. Mechanisms of Development 99:143–148. DOI: https://doi.org/10.1016/S0925-4773(00)00466-4, PMID: 11091082
Takebayashi H, Ohtsuki T, Uchida T, Kawamoto S, Okubo K, Ikenaka K, Takeichi M, Chisaka O, Nabeshima Y. 2002. Non-overlapping expression of Olig3 and Olig2 in the embryonic neural tube. Mechanisms of Development 113:169–174. DOI: https://doi.org/10.1016/S0925-4773(02)00021-7, PMID: 11960707
Vue TY, Aaker J, Taniguchi A, Kazemzadeh C, Skidmore JM, Martin DM, Martin JF, Treier M, Nakagawa Y. 2007. Characterization of progenitor domains in the developing mouse thalamus. The Journal of Comparative Neurology 505:73–91. DOI: https://doi.org/10.1002/cne.21467, PMID: 17729296
Wallace VA. 1999. Purkinje-cell-derived sonic hedgehog regulates granule neuron precursor cell proliferation in the developing mouse cerebellum. Current Biology 9:445–448. DOI: https://doi.org/10.1016/S0960-9822(99) 80195-X, PMID: 10226030
Wang VY, Rose MF, Zoghbi HY. 2005. Math1 expression redefines the rhombic lip derivatives and reveals novel lineages within the brainstem and cerebellum. Neuron 48:31–43. DOI: https://doi.org/10.1016/j.neuron.2005. 08.024
Wechsler-Reya RJ, Scott MP. 1999. Control of neuronal precursor proliferation in the cerebellum by sonic hedgehog. Neuron 22:103–114. DOI: https://doi.org/10.1016/S0896-6273(00)80682-0, PMID: 10027293
West MJ. 2012. Estimating volume in biological structures. Cold Spring Harbor Protocols 2012:1129–1139. DOI: https://doi.org/10.1101/pdb.top071787, PMID: 23118357
Wingate RJ, Hatten ME. 1999. The role of the rhombic lip in avian cerebellum development. Development 126: 4395–4404.
Yamada M, Seto Y, Taya S, Owa T, Inoue YU, Inoue T, Kawaguchi Y, Nabeshima Y, Hoshino M. 2014. Specification of spatial identities of cerebellar neuron progenitors by ptf1a and atoh1 for proper production of GABAergic and glutamatergic neurons. The Journal of Neuroscience 34:4786–4800. DOI: https://doi.org/10. 1523/JNEUROSCI.2722-13.2014, PMID: 24695699
Yeung J, Ha TJ, Swanson DJ, Goldowitz D. 2016. A novel and multivalent role of Pax6 in cerebellar development. Journal of Neuroscience 36:9057–9069. DOI: https://doi.org/10.1523/JNEUROSCI.4385-15. 2016, PMID: 27581449
Zechner D, Müller T, Wende H, Walther I, Taketo MM, Crenshaw EB, Treier M, Birchmeier W, Birchmeier C. 2007. Bmp and wnt/beta-catenin signals control expression of the transcription factor Olig3 and the specification of spinal cord neurons. Developmental Biology 303:181–190. DOI: https://doi.org/10.1016/j. ydbio.2006.10.045, PMID: 17150208
Zervas M, Millet S, Ahn S, Joyner AL. 2004. Cell behaviors and genetic lineages of the mesencephalon and rhombomere 1. Neuron 43:345–357. DOI: https://doi.org/10.1016/j.neuron.2004.07.010, PMID: 15294143