Radial glial progenitor cells exhibit bidirectional cell cycle–dependent nuclear oscillations. The purpose and underlying mechanism of this unusual 'interkinetic nuclear migration' are poorly understood. We investigated the basis for this behavior by live imaging of nuclei, centrosomes and microtubules in embryonic rat brain slices, coupled with the use of RNA interference (RNAi) and the myosin inhibitor blebbistatin. We found that nuclei migrated independent of centrosomes and unidirectionally away from or toward the ventricular surface along microtubules, which were uniformly oriented from the ventricular surface to the pial surface of the brain. RNAi directed against cytoplasmic dynein specifically inhibited nuclear movement toward the apical surface. An RNAi screen of kinesin genes identified Kif1a, a member of the kinesin-3 family, as the motor for basally directed nuclear movement. These observations provide direct evidence that kinesins are involved in nuclear migration and neurogenesis and suggest that a cell cycle–dependent switch between distinct microtubule motors drives interkinetic nuclear migration.
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We thank K.H. Bremner for assistance in surgical procedures, X. Wang for help in molecular cloning, G. Kreitzer for reagents and X. Wang, G. Kreitzer, W. Gruber, J. Canman and S. Cappello for helpful comments and suggestions. This work was supported by US National Institutes of Health grant HD40182 to R.B.V. and a New York State Spinal Cord Injury Research Board fellowship to J.-W.T.
The authors declare no competing financial interests.
Supplementary Figures 1–4 and Supplementary Tables 1–3 (PDF 2777 kb)
Dynamics of normal interkinetic nuclear migration of radial glial progenitors. (MOV 1862 kb)
Centrosomal dynamics in radial glial prenitors throughout cell cycle. (MOV 2729 kb)
Centrosomal dynamics in radial glial progenitors throughout cell cycle. (MOV 3993 kb)
Centrosomal dynamics in radial glial progenitors throughout cell cycle. (MOV 772 kb)
EB3 movements in premitotic radial glial progenitors. (MOV 360 kb)
EB3 movements in premitotic radial glial progenitors. (MOV 464 kb)
EB3 movements in radial glial progenitors during mitosis. (MOV 1259 kb)
EB3 movements in radial glial progenitors during cytokinesis. (MOV 637 kb)
EB3 movements in radial glial progenitors after symmetric cell division. (MOV 601 kb)
EB3 movements in radial glial progenitors after asymmetric cell division. (MOV 1672 kb)
Behavior of a radial glial progenitor after 5 days of dynein HC RNAi. (MOV 980 kb)
Behavior of a radial glial progenitor after 1.5 days of dynamytin overepxression. (MOV 1305 kb)
Behavior of a radial glial progenitor after 3 days of dynein HC RNAi. (MOV 226 kb)
Basally directed nuclear movment of a pair of radial glial progenitor progeny after 3 days of dynein HC RNAi. (MOV 267 kb)
Basally directed nuclear movement of a radial glial progenitor after 2 days of myosin IIB RNAi. (MOV 349 kb)
Basally directed nuclear movement of a radial glial progenitor after 2 days of myosin IIB control RNAi. (MOV 1097 kb)
Apically directed nuclear movement of a blebbistatin treated radial glial progenitor cell. (MOV 319 kb)
Basally directed nuclear movement of a blebbistatin treated radial glial progenitor cell. (MOV 163 kb)
Nuclear movement of a blebbistatin treated mitotic radial glial progenitor cell. (MOV 280 kb)
Basally directed nuclear movement of a radial glial progenitor cells after 2 days of RNAi using scrambled KIF1A sequence. (MOV 290 kb)
Failure in basally directed nuclear movement of a postmitotic radial glial progenitor cell after 2 days of KIF1A RNAi. (MOV 123 kb)
Differential effects following 2 days of KIF1A RNAi on postmitotic progeny of asymmetrically divided radial glial progenitor cell. (MOV 389 kb)
Failure of basally directed nuclear movement in postmitotic KIF1A shRNA-/DsRed-co-expressing radial glial progenitor cell. (MOV 72 kb)
Rescue of basally directed nuclear movement in KIF1A shRNA-/DsRed-human KIF1A-co-expressing radial glial progenitor cell. (MOV 251 kb)
Almost normal bidirectional nuclear migration behavior in KIF3A shRNA-expressing radial glial progenitor cell. (MOV 264 kb)
Normal basally directed nuclear migration in KIF1B shRNA-expressing radial glial progenitor cell. (MOV 923 kb)
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