Institute of Neuroscience Faculty
Professor, Department of Biology
B.S., New College
Ph.D., Stanford University
Generation of neuron and glial diversity in the Drosophila CNS
Asymmetric cell division in Drosophila
Generation of Cell Polarity, Temporal Identity, and Neural Diversity in Drosophila
Chris Doe's lab investigates Drosophila CNS development. Current interests are (1) how stem cell-like neural precursors (neuroblasts) establish cell polarity and divide asymmetrically; (2) how neuroblasts maintain stem cell-like features as they divide to produce differentiating progeny; (3) how transcription factors regulate temporal identity within neuroblast lineages; and (4) the genetic program governing motor neuron specification.
Asymmetric cell division of neural precursors. Drosophila neural precursors (called neuroblasts) repeatedly divide along their apical/basal axis to regenerate an apical neuroblast and bud off a smaller basal daughter cell (called a GMC) that differentiates into a neurons or glia. Normal asymmetric division requires alignment of the mitotic spindle along the apical/basal axis as well as polarized localization of cell fate determinants to the apical or basal poles of the cell -- which allows two molecularly distinct daughter cells to be produced.
We are interested how neuroblasts establish cell polarity, and how cell polarity is used to generate two different cell types at each cell division. Work from our lab and others has identified basally-localized mRNA and proteins (e.g. prospero RNA and Miranda, Prospero, and Numb proteins) as well as apically-localized proteins (e.g. Baz, Par-6, and aPKC). We have done genetic screens to identify new genes involved in apical protein localization, spindle orientation, and basal protein localization, and have identified 12 loci that are required for one or more of these events. A graduate student in the lab, Sarah Siegrist, has developed methods for timelapse imaging of asymmetric neuroblast division both in vivo and in vitro, which is providing new insights into wild type and mutant cell division phenotypes.
Two former graduate students, Chian-Yu Peng and Roger Albertson, have characterized three basal localization mutants, the previously identified "tumor suppressor genes" lethal giant larvae ( lgl ), discs large ( dlg ), and scribble . All three mutants show normal apical protein localization and spindle orientation, but a loss of basal protein targeting. Interestingly, these phenotypes can be suppressed by reducing the level of non-muscle myosin II protein, and mimicked by a pan-myosin inhibitor, leading to a model in which both positive and negative myosins regulate basal transport of Miranda and Numb proteins. A third graduate student, Karsten Siller, is working on the role of the dynactin complex and Lis1 in regulating basal protein targeting and spindle orientation in neuroblasts. Karsten has show that Lis1 is essential for normal asymmetric division (both basal targeting and spindle orientation). His results are likely to aid in our understanding of the human Lissencephaly phenotype, which has yet to be characterized at the cellular level. Our work on cell polarity and asymmetric cell division has been supported by HHMI and the NIH.
Temporal regulation of cell fate within neuroblast cell lineages. Producing the right cells at the right time is essential for normal development, yet it is not well understood how an embryonic precursor cell or a stem cell reproducibly generates a characteristic sequence of different cell types. To begin to study this question, we have done comprehensive cell lineage studies to identify the clone of neurons and glia produced by all 30 different embryonic neuroblasts ( http://www.neuro.uoregon.edu/doelab/lineages/ ), as well as the precise birth-order of all progeny for selected neuroblasts.
We recently showed that nearly all of the 30 different Drosophila neuroblasts in each segment sequentially express the transcription factors Hunchback ö Krüppel ö Pdm ö Castor, raising the possibility of a molecular "clock" for distinguishing GMC birth-order (Isshiki et al., 2001, Cell 106:511). Interestingly, while neuroblast only transiently expressed each gene, the daughter GMCs born during each window of expression maintained expression of that gene as they differentiated. Thus, first-born GMCs maintain Hunchback as they differentiate, whereas second-born GMCs maintain Kruppel as they differentiate. Mutant and misexpression studies show that Hunchback is necessary and sufficient for first-born cell fates, whereas Krüppel is necessary and sufficient for second-born cell fates; we observe this in multiple neuroblast lineages and is independent of the cell type involved. We postulate that Hunchback ö Krüppel ö Pdm ö Castor are "temporal coordinate genes" that act together with "spatial coordinate genes" known to specify each neuroblast identity to uniquely specify the identity of each neuron or glia in the CNS.
More recently, Bret Pearson in the lab has shown that Hunchback has the potential to "restart" the lineage of older neuroblasts, revealing a surprising degree of plasticity in neuroblast developmental potential. Bret has also shown that transient expression of Hunchback can produce long-term heritable specification of first-born cell fate, suggesting that Hunchback-mediated chromatin remodeling may be involved in the specification of neuronal temporal identity, similar to the role of Hunchback in establishing heritable HOX gene expression.
Other questions that we are interested in are: Do Pdm and Castor have similar functions in specifying later-born fates? What regulates the timing of the gene expression "clock" that controls Hunchback ö Krüppel ö Pdm ö Castor? And, do hunchback and Krüppel orthologs have similar functions during vertebrate neurogenesis or hematopoiesis?
Generation of motor neuron identity. A long-term interest of the lab has been to understand how neural diversity is generated. Two graduate students in the lab, Joanne Odden and Mike Layden, are investigating how specific types of motor neurons are produced. Joanne has shown that the Drosophila HB9 homeodomain transcription factor is expressed in a subset of motor neurons that project to the lateral body wall muscles; these are distinct from the pool of Eve+ motor neurons that project to dorsal body wall muscles and from a small pool of motor neurons that project to the ventral-most muscles. RNAi and misexpression experiments show that HB9 is necessary and sufficient for motor neuron targeting to lateral muscles. Mike is working on a pan-neuronal transcription factor called Zfh1. This transcriptional repressor is also expressed in a subset of glia that are associated with motor nerve roots. Mike is currently doing mutant and misexpression analysis of Zfh1 to test its role in motor neuron specification. Additional studies on other transcription factors expressed in some or all motor neurons are ongoing.
Representative Publications - Full List and PDF downloads
Tran, K.D., Miller, M.R., Doe, C.Q., (2010) Recombineering Hunchback identifies two conserved domains required to maintain neuroblast competence and specify early-born neuronal identity. Development 137(9), 1421-1430. | [PDF]
Siller, K.H., Doe, C.Q., (2009) Spindle orientation during asymmetric cell division. Nature Cell Biology April V.11(3): 365-374. | [PDF]
Marcette, J., Hood, I.V., Johnston, C.A., Doe, C.Q., Prehoda, K.E., (2009) Allosteric Control of Regulated Scaffolding in Membrane-Associated Guanylate Kinases. Biochemistry 48, 10014-10019. | [PDF]
Miller, M.R., Robinson, K.J., Cleary, M.D., Doe, C.Q., (2009) TU-tagging: cell type-specific RNA isolation from intact complex tissues. Nature Methods 6, 439-441. | [PDF]
Cabernard, C., Doe, C.Q., (2009) Apical/Basal Spindle Orientation is Required for Neuroblast Homeostasis and Neuronal Differentiation in Drosophila. Developmental Cell 17, 134-141. | [PDF]
Johnston, C.A., Hirono, K., Prehoda, K.E., Doe, C.Q. (2009) Identification of an Aurora-A/PinsLINKER/Dlg Spindle Orientation Pathway using Induced Cell Polarity in S2 Cells. Cell 138, 1150-1163.| [PDF]
Chabu, C, Doe, C.Q., (2009) Twins/PP2A regulates aPKC to control neuroblast cell polarity and self-renewal. Developmental Biology 330, 399-405. | [PDF]
Tran, K.D., Doe, C.Q., (2008) Pdm and Castor close successive temporal identity windows in the NB-3 lineage. Development 135, 3491-3499. | [PDF]
Chabu, C., Doe, C.Q., (2008) Dap160/intersectin binds and activates aPKC to regulate cell polarity an cell cycle progression. Development 135, 2739-2746. | [PDF]
Siller, K.H., Doe, C.Q., (2008) Lis1/dynactin regulates metaphase spindle orientation in Drosophila neuroblasts. Developmental Biology 319, 1-9. | [PDF]
Boone, J.Q., Doe, C.Q., (2008) Identification of Drosophila Type II Neuroblast Lineages Containing Transit Amplifying Ganglion Mother Cells. Developmental Neurobiology 69, 1185-1195. | [PDF]
Doe, C.Q., (2008) Neural stem cells: balancing self-renewal with differentiation. Development 135, 1575-1587. | [PDF]
Zhu, C.C., Boone, J.Q., Jensen, P.A., Hanna, S., Pdemski, L., Locke, J., Doe, C.Q., O'Connor, M., (2008) Drosopila Activin-ÃŸ and the Activin-like product Dawdle function redundantly to regulate proliferation in the larvel brain. Development 135, 513-521. | [PDF]
Atwood, S.X., Chabu, C., Penkert, R.R., Doe, C.Q., Prehoda, K.E., (2007) Cdc42 acts downstream of Bazooka to regulate neuroblast polarity through Par-6-aPKC. Journal of Cell Science 120, 3200-3206. | [PDF]
Nipper, R.W., Siller, K.H., Smith, N.R., Doe, C.Q.. Prehods, K.E., (2007) G alpha i generates multiple Pins activation states to link cortical polarity and spindle orientation in Drosophila neuroblasts. PNAS 104 (36), 14306-14311. | [PDF]
Cabernard, C., Doe, C.Q. (2007) Stem Cell Self-Renewal: Centrsomes on the Move. Current Biology 17: R465-R467. | [PDF]
Rolls, M.M., Satoh, D., Clyne, P.J., Henner, A.L., Uemura, T., Doe, CQ (2007) Polarity and intracellular compartmentalization of Drosophila neurons. Neural Development Apr 30;2(1):7 | [PDF]
Siegrist, S.E., Doe, C.Q. (2007) Microtubule-induced cortical cell polarity. Genes & Development Mar 1;21(5):483-96 | [PDF]
Egger, B.,Boone, J.Q., Stevens, N.R., Brand, A.H., Doe, C.Q. (2007) Regulation of spindle orientation and neural stem cell fate in the Drosophila optic lobe. Neural Development Jan 5;2:1 | [PDF]
Lee, C-Y, Andersen, R.O., Cabernard, C., Manning, L., Tran, K.D., Lanskey, M.J., Bashirullah, A., Doe, C.Q. (2006) Drosophila Aurora-A kinase inhibits neuroblast self-renewal by regulating aPKC/Numb cortical polarity and spindle orientation. Genes & Development 20 (24), 3464-3474 | [PDF]
Doe, C.Q. (2006) Chinmo and neuroblast temporal identity. Cell Oct 20;127(2):254-6 | [PDF]
Grosskortenhaus, R., Robinson, K.J., Doe, C.Q. (2006) Pdm and Castor specify late-born motor neuron identity in the NB7-1 lineage. Genes & Development Sep 15;20(18):2618-27 | [PDF]
Siller, K.H., Cabernard, C., Doe, C.Q. (2006) The NuMA-related Mud protein binds Pins and regulates spindle orientation in Drosophila neuroblasts. Nature Cell Biology Jun;8(6):594-600 | [PDF]
Lee, C-Y., Wilkinson, B.D., Siegrist, S., Wharton, R., Doe, C.Q. (2006) Brat Is a Miranda Cargo Protein that Promotes Neuronal Differentiation and Inhibits Neuroblast Self-Renewal. Developmental Cell 10, 441-449 | [PDF]
Cleary M., Doe C.Q. (2006) Regulation of neuroblast competence: multiple temporal identity factors specify distinct neuronal fates within a single early competence window. Genes & Development 20:429-434 | [PDF]
Layden M.J., Odden J., Schmid A., Garces A., Thor S., Doe CQ, (2006) Zfh1, a somatic motor neuron transcription factor, regulates axon exit from the CNS. Developmental Biology 291:253-263 | [PDF]
Siegrist, S.E., Doe, C.Q. (2006) Extrinsic cues orient the cell division axis in Drosophila embryonic neuroblasts. Development Feb 133(3):529-36 | [PDF]
Lee C-Y., Robinson K., Doe C.Q. (2006) Lgl, Pins and aPKC regulate neuroblast self-renewal versus differentiation. Nature 439(2) 594-598 | [PDF]
Siegrist S., Doe C.Q. (2005) Microtubule-Induced Pins/Gai Cortical Polarity in Drosophila Neuroblasts. Cell 123:1323-1335 | [PDF]
Siller K., Serr M., Steward R., Hays T.S., Doe C.Q. (2005) Live Imaging of Drosophila Brain Neuroblasts Reveals a Role for Lis1/Dynactin in Spindle Assembly and Mitotic Checkpoint Control. Molecular Biology of the Cell 16;5127-5140 | [PDF]
Armstrong, J.A., Sperling, A.S., Deuring, R., Manning, L., Moseley, S.L., Papoulas, O., Piatek, C.I., Doe, C.Q., Tamkun, J.W. (2005) Genetic screens for enhancers of brahma reveal functional interactions between the BRM chromatin-remodeling complex and the delta-notch signal transduction pathway in Drosophila. Genetics Aug 170(4):1761-74 | [PDF]
Grosskortenhaus R, Pearson BJ, Marusich A, Doe C.Q. (2005) Regulation of temporal identity transitions in Drosophila neuroblasts. Developmental Cell. 2005 Feb;8(2):193-202 | [PDF]
Cheesman, S., Layden, M., von Ohlen, T., Doe, C.Q. and Eisen, J.S. (2004) Zebrafish and fly Nkx6 proteins have similar CNS expression patterns and regulate motoneuron formation. Development 131 5221-5232. | [PDF]
Pearson, B., Doe, C.Q. (2004) Specification of temporal identity in the developing nervous system. Annual Review of Cell and Developmental Biology, Vol. 20, pages 619-647. | [PDF]
Albertson, R., Chabu, C., Sheehan, A. and Doe, C.Q. (2003) Scribble protein domain mapping reveals a multistep localization mechanism and domains necessary for establishing cortical polarity. Journal of Cell Science 117, 6061-6070. | [PDF]
Karcavich, R. and Doe, C.Q. (2005) The Drosophila neuroblast 7-3 cell lineage: a model system for studying programmed cell death, Notch/Numb signaling, and sequential specification of GMC identity. The Journal of Comparative Neurology, 481:240-251 | [PDF]
Rolls, M.M. and and Doe, C.Q. Baz, Par-6 and aPKC are polarized in mature neurons, but are not required for in vivo axon specification in Drosophila. Nature Neuroscience., 421, 905-906. | [PDF]
Irion, U., Leptin, M., Siller, K., Fuerstenberg, S., Cai, Y., Doe, C.Q., Chia, W., and Yang, X. (2004) Abstrakt, a DEAD box protein, regulates inscuteable at a post-transcriptional level and is required for asymmetric division of neural and mesodermal progenitors. Current Biology, 14, 138-144. | [PDF]
Isshiki, T. and Doe, C.Q. (2004) Youthfulness of neural progenitors in Drosophila. Cell Cycle, 3, 296-299. | [PDF]
Freeman, M., Delrow, J., Kim, J., Johnson, E., Doe, C.Q. (2003) Unwrapping glial biology: Gcm target genes regulating glial development, diversification, and function. Neuron, 38, 567-580 | [PDF] [Movie 1] [Movie 2]
McDonald, J.A. Fujioka, M., Odden, J.P., Jaynes, J.B., Doe, C.Q. (2003) Specification of motoneuron fate in Drosophila: Integration of positive and negative transcription factor inputs by a minimal eve enhancer. Journal of Neurobiology 57(2), 193-203 | [PDF]
Odden, J., Holbrook, S., and Doe, C.Q. (2002) Drosophila HB9 Is Expressed in a Subset of Motoneurons and Interneurons, Where It Regulates Gene Expression and Axon Pathfinding. The Journal of Neuroscience | [PDF]
Zelhof, A.C., Bao, H., Hardy, R.W., Zhang, B., and Doe, C.Q. Drosophila Amphiphysin is implicated in protein localization and membrane morphogenesis but not in endocytosis., Development 128, 5005-5015. | [PDF]
Isshiki, T., Pearson, B., Holbrook, S., and Doe, C.Q. (2001) Drosophila neuroblasts sequentially express transcription factors which specify the temporal identity of their neuronal progeny. Cell106, 511-521. | [PDF]
Von Ohlen, T. and Doe, C.Q. (2000) Convergence of Dorsal, Dpp and Egfr
signaling pathways subdivides the Drosophila neuroectoderm into
three dorsoventral columns. Dev.
Biol.224, 362-372. | [PDF]
Schmid, A., Chiba, A., and Doe, C.Q. (1999) Clonal analysis of Drosophila embryonic neuroblasts: neural cell types, axon projections and muscle targets. Development 126, 4653-4689. | [PDF] [Data]
Weiss, J., Von Ohlen, T., Mellerick, D., Dressler, G., Doe, C., and Scott, M. (1998) Dorsal-ventral patterning in the Drosophila central nervous system: The intermediate neuroblasts defective homeobox gene specifies intermediate column identity. Genes & Dev. 12, 3591-3602. | [PDF]
McDonald, J.A., Holbrook, S., Isshiki, T., Weiss, J., Doe, C., and Mellerick, D. (1998) Dorsoventral patterning in the Drosophila CNS: the vnd homeobox gene specifies ventral column identity. Genes & Development. 12, 3603-3612. | [PDF]
Srinivasan, S., Peng, C.-Y., Skeath, J.B., Nair, S., Spana, E.P., Lassy, C., Doe, C.Q. (1998) Biochemical analysis of the Prospero protein during asymmetric cell division: cortical Prospero is highly phosphorylated relative to nuclear Prospero. Developmental Biology 204, 478-487. | [PDF]
Broadus, J.B. and Doe, C.Q. (1997) Extrinsic cues, intrinsic cues, and microfilaments regulate asymmetric localization of Prospero, Staufen, and Inscuteable in Drosophila neuroblasts. Current Biology. 7, 827-835. | [PDF]
McDonald, J.A. and Doe, C.Q. (1997) Establishing neuroblast-specific gene expression in the Drosophila CNS: huckebein is activated by wingless and hedgehog and repressed by engrailed and gooseberry. Development 124, 1079-1087. | [PDF]
Chu-LaGraff, Q., Schmid, A., Leidel, J., Bronner, G., Jackle, H., and Doe, C.Q. (1995) huckebein specifies aspects of CNS precursor identity required for motoneuron axon pathfinding. Neuron 15, 1041-1051.
Spana, E., Kopczynski, C., Goodman, C.S. and Doe, C.Q. (1995) Asymmetric localization of numb autonomously determines sibling neuron identity in the Drosophila CNS. Development 121, 3489-3494. | [PDF]
Skeath, J.B., Zhang, Y., Holmgren, R., Carroll, S.B., and Doe, C.Q. (1995) Specification of neuroblast identity in the Drosophila embryonic central nervous system by gooseberry-distal. Nature 376, 427-430. | [PDF]
Broadus, J., Skeath, J.B., Spana, E.P., Bossing, T., Technau, G., Doe, C.Q. (1995) New neuroblast markers and the origin of the aCC/pCC neurons in the Drosophila central nervous system. Mechanisms of Development 53, 393-402 | [PDF]
Chu-LaGraff, Q., Doe, C.Q. (1993) Neuroblast Specification and Formation Regulated by wingless in the Drosophila CNS. Science 261, 1594-1597. | [PDF]