Abstract
Neuronal migration, like the migration of many cell types, requires an extensive rearrangement of cell shape, mediated by changes in the cytoskeleton. The genetic analysis of human brain malformations has identified several biochemical players in this process, including doublecortin (DCX) and LIS1, mutations of which cause a profound migratory disturbance known as lissencephaly ('smooth brain') in humans. Studies in mice have identified additional molecules such as Cdk5, P35, Reelin, Disabled and members of the LDL superfamily of receptors. Understanding the cell biology of these molecules has been a challenge, and it is not known whether they function in related biochemical pathways or in very distinct processes. The last year has seen rapid advances in the biochemical analysis of several such molecules. This analysis has revealed roles for some of these molecules in cytoskeletal regulation and has shown an unexpected conservation of the genetic pathways that regulate neuronal migration in humans and nuclear movement in simple eukaryotic organisms.
Key Points
-
The analysis of human brain malformations has identified several genes required for neuronal migration during development, including doublecortin (DCX) and LIS1. Mutations in these genes cause a profound migratory disturbance known as lissencephaly ('smooth brain'), and recent studies have shown that they encode proteins that are important for cytoskeletal regulation.
-
Autosomal dominant and X-linked forms of lissencephaly have been identified. The LIS1 gene on chromosome 17 is responsible for a large percentage of the autosomal dominant forms, and doublecortin (DCX) is one of the genes responsible for X-linked lissencephaly. In males, DCX mutations produce lissencephaly phenotypes similar to those associated with LIS1 mutations. Heterozygous females show double cortex syndrome, in which a population of neurons arrests halfway between the cortex and the ventricle, forming a subcortical band in the white matter.
-
DCX is a microtubule-associated protein (MAP) that is expressed exclusively in postmitotic neurons and colocalizes with polymerized microtubules. DCX seems to stimulate polymerization and bundling of microtubules, properties that are disrupted by disease-causing mutations.
-
LIS1 encodes a ubiquitously expressed 45 kDa protein with 7 WD40 repeats, which are thought to mediate protein–protein interactions. LIS1 is reported to be a MAP that binds microtubules directly and it is thought to function in microtubule regulation in migrating neurons.
-
Insights into the microtubule-regulatory function of LIS1 have come from studies of its homologue, nudF, in the filamentous fungus Aspergillus nidulans. nudF mutations cause defects in nuclear migration, a microtubule-mediated event. Mutations in nudF and nudA, which encodes the heavy chain of cytoplasmic dynein, share a similar nuclear migration phenotype, and double mutations have the same phenotype as each single mutation. This suggests that dynein and nudF act in the same nuclear migration pathway. It is proposed that they function via destabilization of microtubules during nuclear migration.
-
The Aspergillus NUDE protein is thought to function as a downstream effector of NUDF in regulating nuclear migration. In mammals, two nudE homologues, mNudE and NUDEL, were cloned from LIS1 two-hybrid screens. They are similar in size and structural organization and both interact with LIS1 through a structurally similar amino-terminal region. mNudE interacts with multiple centrosomal proteins and might act as a structural and functional scaffold for the microtubule-organizing centre (MTOC). NUDEL binds directly to dynein and regulates its subcellular distribution. It is enriched in the processes of mature neurons, suggesting additional roles in axon transport and neurite outgrowth.
-
LIS1 physically interacts and forms complexes with dynein in mammals, and its overexpression induces spindle mis-orientation and mitotic arrest at M-phase, suggesting that LIS1–dynein interactions might be important for cell division. It is also suggested that LIS1–dynein interactions affect the translocation of the neuronal cell body during migration.
-
A model is proposed for LIS1- and DCX-mediated neuronal migration. Neurons receive migration signals and microtubules extend into the leading process. Then, LIS1 is modified and is recruited to the MTOC, where it binds to mNudE and/or NUDEL. This interaction reduces nucleation and polymerization of microtubules at the minus end, possibly by regulating the γ-tubulin complex. As a result, the microtubules shorten at the minus end and the nucleus is pulled towards the leading edge of the migrating neuron by microtubule-based motors, such as dyneins.
-
Other factors that underlie neuronal migration and might interact with the lissencephaly gene products have been identified. For example, the Cdk5 kinase is a candidate for regulating LIS1/DCX phosphorylation and function. Mutations in other genes, such as Reelin, Disabled, ApoE2R and VLDLR also cause defects in cortical neuronal migration. These all encode components of the Reelin signalling pathway, which has been implicated in regulation of the microtubule cytoskeleton.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Jellinger, K. & Rett, A. Agyria–pachygyria (lissencephaly syndrome). Neuropadiatrie 7, 66–91 (1976).
Alvarez, L. A. et al. Miller–Dieker syndrome: a disorder affecting specific pathways of neuronal migration. Neurology 36, 489–493 (1986).
Kuchelmeister, K., Bergmann, M. & Gullotta, F. Neuropathology of lissencephalies. Childs Nerv. Syst. 9, 394–399 (1993).
Rakic, P. Neuronal migration and contact guidance in the primate telencephalon. Postgrad. Med. J. 54, 25–40 (1978).
Gleeson, J. G. & Walsh, C. A. Neuronal migration disorders: from genetic diseases to developmental mechanisms. Trends Neurosci. 23, 352–359 (2000).
Dobyns, W. B., Reiner, O., Carrozzo, R. & Ledbetter, D. H. Lissencephaly: a human brain malformation associated with deletion of the LIS1 gene located at chromosome 17p13. JAMA 270, 2838–2842 (1993).
Lo Nigro, C. et al. Point mutations and an intragenic deletion in LIS1, the lissencephaly causative gene in isolated lissencephaly sequence and Miller–Dieker syndrome. Hum. Mol. Genet. 6, 157–164 (1997).
Reiner, O. et al. Isolation of a Miller–Dieker lissencephaly gene containing G protein β-subunit-like repeats. Nature 364, 717–721 (1993).
Hirotsune, S. et al. Graded reduction of Pafah1b1 (Lis1) activity results in neuronal migration defects and early embryonic lethality. Nature Genet. 19, 333–339 (1998).
des Portes, V. et al. Doublecortin is the major gene causing X-linked subcortical laminar heterotopia (SCLH). Hum. Mol. Genet. 7, 1063–1070 (1998).
Gleeson, J. G. et al. Doublecortin, a brain-specific gene mutated in human X-linked lissencephaly and double cortex syndrome, encodes a putative signaling protein. Cell 92, 63–72 (1998).
Gleeson, J. G., Lin, P. T., Flanagan, L. A. & Walsh, C. A. Doublecortin is a microtubule-associated protein and is expressed widely by migrating neurons. Neuron 23, 257–271 (1999).
Francis, F. et al. Doublecortin is a developmentally regulated, microtubule-associated protein expressed in migrating and differentiating neurons. Neuron 23, 247–256 (1999).
Horesh, D. et al. Doublecortin, a stabilizer of microtubules. Hum. Mol. Genet. 8, 1599–1610 (1999).References 12 – 14 describe the functional characterization of DCX protein as a MAP, and suggest that DCX functions in neuronal migration by stabilizing microtubules.
Geyer, M. & Wittinghofer, A. GEFs, GAPs, GDIs and effectors: taking a closer (3D) look at the regulation of Ras-related GTP-binding proteins. Curr. Opin. Struct. Biol. 7, 786–792 (1997).
Nassar, N. et al. The 2. 2 A crystal structure of the Ras-binding domain of the serine/threonine kinase c-Raf1 in complex with Rap1A and a GTP analogue. Nature 375, 554–560 (1995).
Taylor, K. R., Holzer, A. K., Bazan, J. F., Walsh, C. A. & Gleeson, J. G. Patient mutations in doublecortin define a repeated tubulin-binding domain. J. Biol. Chem. 275, 34442–34450 (2000).
Drechsel, D. N., Hyman, A. A., Cobb, M. H. & Kirschner, M. W. Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau. Mol. Biol. Cell 3, 1141–1154 (1992).
Kaech, S., Ludin, B. & Matus, A. Cytoskeletal plasticity in cells expressing neuronal microtubule-associated proteins. Neuron 17, 1189–1199 (1996).
Garner, C. C., Garner, A., Huber, G., Kozak, C. & Matus, A. Molecular cloning of microtubule-associated protein 1 (MAP1A) and microtubule-associated protein 5 (MAP1B): identification of distinct genes and their differential expression in developing brain. J. Neurochem. 55, 146–154 (1990).
Schoenfeld, T. A., McKerracher, L., Obar, R. & Vallee, R. B. MAP 1A and MAP 1B are structurally related microtubule associated proteins with distinct developmental patterns in the CNS. J. Neurosci. 9, 1712–1730 (1989).
Chen, J., Kanai, Y., Cowan, N. J. & Hirokawa, N. Projection domains of MAP2 and tau determine spacings between microtubules in dendrites and axons. Nature 360, 674–677 (1992).
Sapir, T. et al. Doublecortin mutations cluster in evolutionarily conserved functional domains. Hum. Mol. Genet. 9, 703–712 (2000).
Rakic, P., Knyihar-Csillik, E. & Csillik, B. Polarity of microtubule assemblies during neuronal cell migration. Proc. Natl Acad. Sci. USA 93, 9218–9222 (1996).
Lin, P. T., Gleeson, J. G., Corbo, J. C., Flanagan, L. & Walsh, C. A. DCAMKL1 encodes a protein kinase with homology to doublecortin that regulates microtubule polymerization. J. Neurosci. 20, 9152–9161 (2000).
Burgess, H. A. & Reiner, O. Doublecortin-like kinase is associated with microtubules in neuronal growth cones. Mol. Cell. Neurosci. 16, 529–541 (2000).
Garcia-Higuera, I. et al. Folding of proteins with WD-repeats: comparison of six members of the WD-repeat superfamily to the G protein β subunit. Biochemistry 35, 13985–13994 (1996).
Hattori, M., Adachi, H., Tsujimoto, M., Arai, H. & Inoue, K. Miller–Dieker lissencephaly gene encodes a subunit of brain platelet-activating factor acetylhydrolase. Nature 370, 216–218 (1994).
Brunati, A. M. et al. The spleen protein-tyrosine kinase TPK-IIB is highly similar to the catalytic domain of p72syk. Eur. J. Biochem. 240, 400–407 (1996).
Kurosaki, T. Molecular dissection of B cell antigen receptor signaling (review). Int. J. Mol. Med. 1, 515–527 (1998).
Sapir, T., Elbaum, M. & Reiner, O. Reduction of microtubule catastrophe events by LIS1, platelet-activating factor acetylhydrolase subunit. EMBO J. 16, 6977–6984 (1997).Presented the observation of the colocalization of LIS1 with the microtubule network, its direct binding to polymerized microtubules and its activity in reducing microtubule catastrophe events in vitro.
Oakley, B. R. & Morris, N. R. Nuclear movement is β-tubulin-dependent in Aspergillus nidulans. Cell 19, 255–262 (1980).
Morris, N. R., Lai, M. H. & Oakley, C. E. Identification of a gene for α-tubulin in Aspergillus nidulans. Cell 16, 437–442 (1979).
Oakley, C. E. & Oakley, B. R. Identification of γ-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans. Nature 338, 662–664 (1989).
Xiang, X., Roghi, C. & Morris, N. R. Characterization and localization of the cytoplasmic dynein heavy chain in Aspergillus nidulans. Proc. Natl Acad. Sci. USA 92, 9890–9894 (1995).
Robb, M. J., Wilson, M. A. & Vierula, P. J. A fungal actin-related protein involved in nuclear migration. Mol. Gen. Genet. 247, 583–590 (1995).
Xiang, X., Osmani, A. H., Osmani, S. A., Xin, M. & Morris, N. R. NudF, a nuclear migration gene in Aspergillus nidulans, is similar to the human LIS-1 gene required for neuronal migration. Mol. Biol. Cell 6, 297–310 (1995).
Willins, D. A., Liu, B., Xiang, X. & Morris, N. R. Mutations in the heavy chain of cytoplasmic dynein suppress the nudF nuclear migration mutation of Aspergillus nidulans. Mol. Gen. Genet. 255, 194–200 (1997).
Willins, D. A., Xiang, X. & Morris, N. R. An α tubulin mutation suppresses nuclear migration mutations in Aspergillus nidulans. Genetics 141, 1287–1298 (1995).
Efimov, V. P. & Morris, N. R. The LIS1-related NUDF protein of Aspergillus nidulans interacts with the coiled-coil domain of the NUDE/RO11 protein. J. Cell Biol. 150, 681–688 (2000).
Feng, Y. et al. LIS1 regulates CNS lamination by interacting with mNudE, a central component of the centrosome. Neuron 28, 665–679 (2000).Describes the functional characterization of mNudE and LIS1–NUDE interactions. Showed that mNudE is a potential central scaffold of the microtubule-organizing centre, and that LIS1–NUDE interactions are required for the lamination of anterior CNS structures in Xenopus embryos.
Sasaki, S. et al. A LIS1/NUDEL/cytoplasmic dynein heavy chain complex in the developing and adult nervous system. Neuron 28, 681–696 (2000).Presented evidence that LIS1 and NUDEL form complexes with dynein in the mammalian CNS, and that LIS1 and NUDEL regulate the cellular localization of dynein. It also showed that NUDEL is an in vitro substrate of Cdk5.
Niethammer, M. et al. NUDEL is a novel Cdk5 substrate that associates with LIS1 and cytoplasmic dynein. Neuron 28, 697–711 (2000).Demonstrated that NUDEL interacts with cytoplasmic dynein, and is a phosphoprotein in mammalian brain and a substrate of Cdk5. The distribution of NUDEL is regulated by Cdk5 activity.
Kitagawa, M. et al. Direct association of LIS1, the lissencephaly gene product, with a mammalian homologue of a fungal nuclear distribution protein, rNUDE. FEBS Lett. 479, 57–62 (2000).
Sweeney, K. J., Prokscha, A. & Eichele, G. NudE-L, a novel Lis1-interacting protein, belongs to a family of vertebrate coiled-coil proteins. Mech. Dev. 101, 21–33 (2001).
Faulkner, N. E. et al. A role for the lissencephaly gene LIS1 in mitosis and cytoplasmic dynein function. Nature Cell Biol. 2, 784–791 (2000).Demonstrated that LIS1 interacts with dynein and dynactin, and that overexpression of LIS1 in cultured mammalian cells interferes with mitotic progression and leads to spindle misorientation. Suggests that a LIS1–dynein interaction regulates cell division.
Smith, D. S. et al. Regulation of cytoplasmic dynein behaviour and microtubule organization by mammalian Lis1. Nature Cell Biol. 2, 767–775 (2000).Identified that LIS1 forms complexes with cytoplasmic dynein/dynactin, and presented experimental evidence suggesting that LIS1 regulates cytoplasmic dynein distribution.
Liu, Z., Xie, T. & Steward, R. Lis1, the Drosophila homolog of a human lissencephaly disease gene, is required for germline cell division and oocyte differentiation. Development 126, 4477–4488 (1999).
Swan, A., Nguyen, T. & Suter, B. Drosophila Lissencephaly-1 functions with Bic-D and dynein in oocyte determination and nuclear positioning. Nature Cell Biol. 1, 444–449 (1999).
Liu, Z., Steward, R. & Luo, L. Drosophila Lis1 is required for neuroblast proliferation, dendritic elaboration and axonal transport. Nature Cell Biol. 2, 776–783 (2000).By a mosaic analysis using a Lis1 null mutation, this paper showed that Lis1 is required for neuroblast proliferation and axonal transport. It also showed that neurons containing a mutated cytoplasmic dynein heavy chain (Dhc64C) exhibit phenotypes similar to Lis1 mutants.
Komuro, H. & Rakic, P. Intracellular Ca2+ fluctuations modulate the rate of neuronal migration. Neuron 17, 275–285 (1996).
Hatten, M. E. Central nervous system neuronal migration. Annu. Rev. Neurosci. 22, 511–539 (1999).
Rivas, R. J. & Hatten, M. E. Motility and cytoskeletal organization of migrating cerebellar granule neurons. J. Neurosci. 15, 981–989 (1995).
Nadarajah, B., Brunstrom, J. E., Grutzendler, J., Wong, R. O. & Pearlman, A. L. Two modes of radial migration in early development of the cerebral cortex. Nature Neurosci. 4, 143–150 (2001).
Caspi, M., Atlas, R., Kantor, A., Sapir, T. & Reiner, O. Interaction between LIS1 and doublecortin, two lissencephaly gene products. Hum. Mol. Genet. 9, 2205–2213 (2000).
Sapir, T., Cahana, A., Seger, R., Nekhai, S. & Reiner, O. LIS1 is a microtubule-associated phosphoprotein. Eur. J. Biochem. 265, 181–188 (1999).
Peters, J. D., Furlong, M. T., Asai, D. J., Harrison, M. L. & Geahlen, R. L. Syk, activated by cross-linking the B-cell antigen receptor, localizes to the cytosol where it interacts with and phosphorylates α-tubulin on tyrosine. J. Biol. Chem. 271, 4755–4762 (1996).
Faruki, S., Geahlen, R. L. & Asai, D. J. Syk-dependent phosphorylation of microtubules in activated B-lymphocytes. J. Cell Sci. 113, 2557–2565 (2000).
Stukenberg, P. T. et al. Systematic identification of mitotic phosphoproteins. Curr. Biol. 7, 338–348 (1997).
Ohshima, T. et al. Targeted disruption of the cyclin-dependent kinase 5 gene results in abnormal corticogenesis, neuronal pathology and perinatal death. Proc. Natl Acad. Sci. USA 93, 11173–11178 (1996).
Chae, T. et al. Mice lacking p35, a neuronal specific activator of Cdk5, display cortical lamination defects, seizures, and adult lethality. Neuron 18, 29–42 (1997).
Sobue, K. et al. Interaction of neuronal Cdc2-like protein kinase with microtubule-associated protein tau. J. Biol. Chem. 275, 16673–16680 (2000).
Patrick, G. N. et al. Conversion of p35 to p25 deregulates Cdk5 activity and promotes neurodegeneration. Nature 402, 615–622 (1999).
Ahlijanian, M. K. et al. Hyperphosphorylated tau and neurofilament and cytoskeletal disruptions in mice overexpressing human p25, an activator of cdk5. Proc. Natl Acad. Sci. USA 97, 2910–2915 (2000).
Trommsdorff, M. et al. Reeler/Disabled-like disruption of neuronal migration in knockout mice lacking the VLDL receptor and ApoE receptor 2. Cell 97, 689–701 (1999).
Ware, M. L. et al. Aberrant splicing of a mouse disabled homolog, mdab1, in the scrambler mouse. Neuron 19, 239–249 (1997).
Fox, J. W. et al. Mutations in filamin 1 prevent migration of cerebral cortical neurons in human periventricular heterotopia. Neuron 21, 1315–1325 (1998).
Dulabon, L. et al. Reelin binds α3β1 integrin and inhibits neuronal migration. Neuron 27, 33–44 (2000).
D'Arcangelo, G. et al. A protein related to extracellular matrix proteins deleted in the mouse mutant reeler. Nature 374, 719–723 (1995).
Caviness, V. S. Jr Neocortical histogenesis in normal and reeler mice: a developmental study based upon [3H]thymidine autoradiography. Brain Res. 256, 293–302 (1982).
Pinto-Lord, M. C., Evrard, P. & Caviness, V. S. Jr Obstructed neuronal EM analysis. Brain Res. 256, 379–393 (1982).
Hong, S. E. et al. Autosomal recessive lissencephaly with cerebellar hypoplasia is associated with human RELN mutations. Nature Genet. 26, 93–96 (2000).
Howell, B. W., Gertler, F. B. & Cooper, J. A. Mouse disabled (mDab1): a Src binding protein implicated in neuronal development. EMBO J. 16, 121–132 (1997).
Sheldon, M. et al. Scrambler and yotari disrupt the disabled gene and produce a reeler-like phenotype in mice. Nature 389, 730–733 (1997).
D'Arcangelo, G. et al. Reelin is a ligand for lipoprotein receptors. Neuron 24, 471–479 (1999).
Hiesberger, T. et al. Direct binding of Reelin to VLDL receptor and ApoE receptor 2 induces tyrosine phosphorylation of disabled-1 and modulates tau phosphorylation. Neuron 24, 481–489 (1999).
Nikolic, M., Chou, M. M., Lu, W., Mayer, B. J. & Tsai, L. H. The p35/Cdk5 kinase is a neuron-specific Rac effector that inhibits Pak1 activity. Nature 395, 194–198 (1998).
Acknowledgements
The authors gratefully acknowledge the support of the National Institutes of Health. Research in the authors' laboratory was also supported by a grant from the March of Dimes.
Author information
Authors and Affiliations
Glossary
- HYPOMORPHIC MUTATION
-
A mutation that does not eliminate the wild-type function of a gene and gives a less severe phenotype than a loss-of-function mutation.
- PROJECTION DOMAIN
-
Microtubule-associated proteins are usually organized into two domains: the microtubule-binding domain and a projection domain. By electron microscopy, the projection domain can be seen as a filamentous arm extending from the wall of the microtubule; this arm can bind to membranes, intermediate filaments or other microtubules. Its length controls the spacing between microtubules.
- CATASTROPHE
-
The abrupt transition of a microtubule — a dynamic polymer — from growing phase to shortening phase.
- ORTHOLOGUES
-
Genes belonging to different organisms that carry out similar functions.
- TWO-HYBRID SCREEN
-
A system used to determine the existence of direct interactions between proteins. It involves the use of plasmids that encode two hybrid proteins; one of them is fused to the GAL4 DNA-binding domain and the other one is fused to the GAL4 activation domain. The two proteins are expressed together in yeast and, if they interact, the resulting complex will drive the expression of a reporter gene, commonly β-galactosidase.
- CELL CORTEX
-
The part of the microtrabecular lattice that lies under the cell membrane.
- KINETOCHORE
-
Specialized assembly of proteins that binds to the centromeric region of the chromosome.
- MINUS END
-
The end of the microtubule that anchors at the microtubule-organizing centre. It is also the end that assembles more slowly.
- SURFACE PLASMON RESONANCE
-
An instrumental biosensor or device that detects alterations in the optical evanescent waves resulting from small changes in refractive index at the interface between the sample and the device. The instrument can measure biomolecular interactions in real time and allows the detailed analysis of the resultant signals.
Rights and permissions
About this article
Cite this article
Feng, Y., Walsh, C. Protein–Protein interactions, cytoskeletal regulation and neuronal migration . Nat Rev Neurosci 2, 408–416 (2001). https://doi.org/10.1038/35077559
Issue Date:
DOI: https://doi.org/10.1038/35077559
This article is cited by
-
Dynamics, nanomechanics and signal transduction in reelin repeats
Scientific Reports (2019)
-
α-Synuclein is a Novel Microtubule Dynamase
Scientific Reports (2016)
-
FoxP2 protein levels regulate cell morphology changes and migration patterns in the vertebrate developing telencephalon
Brain Structure and Function (2016)
-
ALK5-dependent TGF-β signaling is a major determinant of late-stage adult neurogenesis
Nature Neuroscience (2014)
-
No effect of running and laboratory housing on adult hippocampal neurogenesis in wild caught long-tailed wood mouse
BMC Neuroscience (2009)