Dual subcellular roles for LIS1 and dynein in radial neuronal migration in live brain tissue.
During brain development, neural precursor cells migrate along radial glial fibers to populate the neocortex. RNA interference (RNAi) of the lissencephaly gene LIS1 (also known as PAFAH1b1) inhibits somal movement but not process extension of neural precursors in live brain slices. Here we report imaging of the subcellular events accompanying neural precursor migration and the effects of LIS1, cytoplasmic dynein and myosin II inhibition. Centrosomes move continuously and often far in advance of nuclei, which show extreme saltatory behavior. LIS1 and dynein RNAi inhibit centrosomal and nuclear movement independently, whereas myosin II inhibition blocks only nuclear translocation. Imaging of the microtubule end-binding protein 3 (EB3) reveals a centrosome-centered array of microtubules in live neural precursors under all conditions examined. Dynein is concentrated both at a swelling in the leading process reported to initiate each migratory cycle and in the soma. Thus, dynein pulls on the microtubule network from the swelling. The nucleus is transported along the trailing microtubules by dynein assisted by myosin II.
Tsai
Jin-Wu
JW
Integrated Program in Cellular, Molecular and Biophysical Studies, Columbia University, 630 W 168th Street, New York, New York 10032, USA.
Bremner
K Helen
KH
Vallee
Richard B
RB
D000818:Animals; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D002540:Cerebral Cortex / Q000166:cytology*; D004398:Dyneins / Q000737:chemistry / Q000502:physiology*; D018274:Electroporation / Q000379:methods; D004622:Embryo, Mammalian; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D024462:Myosin Type II / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D009924:Organ Culture Techniques; D051381:Rats; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology; D013997:Time Factors
D006576:Heterocyclic Compounds, 4 or More Rings; D008164:Luminescent Proteins; C403664:MAPRE3 protein, human; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D016376:Oligonucleotides, Antisense; C516550:Pafah1b1 protein, rat; C472645:blebbistatin; D024462:Myosin Type II; D004398:Dyneins