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Impaired Neuronal Positioning and Dendritogenesis in the Neocortex after Cell-Autonomous Dab1 Suppression

Olson, Eric C., Kim, Seonhee, Walsh, Christopher A.·2006·The Journal of Neuroscience
DOI 10.1523/jneurosci.3000-05.2006
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Abstract

Reelin and Disabled 1 (Dab1) are essential for positioning migrating neurons in the developing neocortex. Cell-autonomous RNA interference-mediated suppression of Dab1 in migrating neurons destined for layer 2/3 shifted the median position of these cells to deeper positions within the cortex. At the time of migration arrest [embryonic day 20 (E20) to E21], Dab1-suppressed cells were underrepresented in the upper ∼40 μm of the cortex compared with controls, suggesting that Dab1 is essential for somal translocation through the cell-dense cortical plate. Closer examination of the morphology of Dab1-suppressed neurons at E20 revealed simplified leading processes that are less likely to contact the marginal zone (MZ), in which high levels of Reelin are expressed. Examination of Dab1-suppressed cells 3 d later (postnatal day 2) revealed simplified dendrites that are also less likely to contact the MZ. These data reveal a cell-autonomous role of Dab1 in dendritogenesis in the neocortex and suggest that remodeling of the leading process of a migrating neuron into a nascent dendrite by Reelin/Dab1 signaling plays an important role in cell positioning.

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Figures

Figure 1 Expression of layer-appropriate markers in electroporated cells. P0 coronal sections of cerebral cortex 5 d after E16 pCAG-GFP electroporations show appropriate expression of layer-specific marker. GFP + cells in the P0 cortex immunolocalize with the POU domain transcription factor Brn1, a marker of layer 2-5 cells ( A , B ), and the Cut-related transcription factor Cux1, a markeroflayer 2- 4 cells ( C , D ) in the mouse cortex. In addition, GFP + cells are positive for DCX, a marker of immature postmitotic neurons ( E , F ). B , D , and F are single confocal sections that have been digitally zoomed three times, whereas A , C , and E are flattened z -series through 10 𝛍 mof tissue. Scale bars: E , 50 𝛍 m; F , 20 𝛍 m.
Figure 1 Expression of layer-appropriate markers in electroporated cells. P0 coronal sections of cerebral cortex 5 d after E16 pCAG-GFP electroporations show appropriate expression of layer-specific marker. GFP + cells in the P0 cortex immunolocalize with the POU domain transcription factor Brn1, a marker of layer 2-5 cells ( A , B ), and the Cut-related transcription factor Cux1, a markeroflayer 2- 4 cells ( C , D ) in the mouse cortex. In addition, GFP + cells are positive for DCX, a marker of immature postmitotic neurons ( E , F ). B , D , and F are single confocal sections that have been digitally zoomed three times, whereas A , C , and E are flattened z -series through 10 𝛍 mof tissue. Scale bars: E , 50 𝛍 m; F , 20 𝛍 m.CC-BY-NC-SA · From the PDF
Figure 2 In vivo morphological changes in migrating neurons in contact with the marginal zone. Time course of cell migration after E16 electroporation. A , At E17, 1 d after electroporation, GFP + cells in sections of the neocortex are found primarily in the ventricular zone (VZ). By E19 ( B ) and E20 ( C ), GFP + cells have entered the cortex and are approaching the MZ. D , By P0, typically 5 d after electroporation, GFP + cells are found in a tight layer underneath the MZ. E , F , Examples of the first group of GFP + cells contacting the MZ from an E19 section of E16 pCAG-GFP-electroporated (green) and E16 bromodeoxyuridine (BrdU)-labeled (red) cortex. E , The white box identifies the migrating cells at the leading edge of the GFP + cells that are examined at higher magnification in F . The cell on the left has a clear branch (yellow arrow) approximately two cell diameters above the CP, but the cell on the right is not branched. G , Arrows identify the BrdU + nuclei from the cells shown in F . H , A higher percentage of cells show leading process branching if the process was in the MZ as opposed to the cortex (CP) (*** p < 0.001, 𝛘 2 test; n = 30 leading processes in MZ; n = 25 leading processes in CP). I , Histogram of the location of the principal branch point with respect to the CP-MZ boundary (red line, x = 0). The x -axis is in nuclear diameters (7 𝛍 m). A majority of branch points in MZ-contacting cells werefoundinthefirst two cell diameters above the CP-MZ boundary. The leading cell population ( n = 55cells) was derived from z -series from four E16 electroporated brains fixed at E19 ( n = 1) or E20 ( n = 3). Scale bars: D , 500 𝛍 m; G , 25 𝛍 m.
Figure 2 In vivo morphological changes in migrating neurons in contact with the marginal zone. Time course of cell migration after E16 electroporation. A , At E17, 1 d after electroporation, GFP + cells in sections of the neocortex are found primarily in the ventricular zone (VZ). By E19 ( B ) and E20 ( C ), GFP + cells have entered the cortex and are approaching the MZ. D , By P0, typically 5 d after electroporation, GFP + cells are found in a tight layer underneath the MZ. E , F , Examples of the first group of GFP + cells contacting the MZ from an E19 section of E16 pCAG-GFP-electroporated (green) and E16 bromodeoxyuridine (BrdU)-labeled (red) cortex. E , The white box identifies the migrating cells at the leading edge of the GFP + cells that are examined at higher magnification in F . The cell on the left has a clear branch (yellow arrow) approximately two cell diameters above the CP, but the cell on the right is not branched. G , Arrows identify the BrdU + nuclei from the cells shown in F . H , A higher percentage of cells show leading process branching if the process was in the MZ as opposed to the cortex (CP) (*** p < 0.001, 𝛘 2 test; n = 30 leading processes in MZ; n = 25 leading processes in CP). I , Histogram of the location of the principal branch point with respect to the CP-MZ boundary (red line, x = 0). The x -axis is in nuclear diameters (7 𝛍 m). A majority of branch points in MZ-contacting cells werefoundinthefirst two cell diameters above the CP-MZ boundary. The leading cell population ( n = 55cells) was derived from z -series from four E16 electroporated brains fixed at E19 ( n = 1) or E20 ( n = 3). Scale bars: D , 500 𝛍 m; G , 25 𝛍 m.CC-BY-NC-SA · From the PDF
Figure 3 Suppression of Dab1 protein expression by RNAi directed against Dab1 mRNA. A , HEK293 cells cotransfected with pSuper.retro.neo plasmids directed against Dab1 (RVC + RVD) and with a plasmid encoding Dab1::GFP show reduced the amount of Dab1::GFPexpressioncomparedwithcontroltransfection(RVNwithDab1::GFP).Aplasmidencodingtheredfluorescentprotein DsRed (pDsRed) was included in all transfections as a transfection efficiency control. Three independent transfections of HEK293 cell cultures were performed to test Dab1::GFP suppression by RVN, RVC, and RVD. Green to red fluorescence ratios within regions of interest was 1.05 ± 0.25 for RVN-treated cultures ( n = 3) compared with 0.20 ± 0.23 for RVC + RVD-treated cultures ( n = 3) (means ± SD; unpaired t test, p = 0.012; df = 4). B , Western blot analysis of whole-cell lysates from HEK293 cell transfections confirms Dab1::GFP suppression after probing with anti-Dab1 antisera (B3) or with an anti-GFP antibody. Reprobing the immunoblot with an anti𝛂 -tubulin antibody confirms approximately equal loading of whole-cell lysates into each lane. C , Suppression of endogenous Dab1 in primary cortical cultures after electroporation of RNAi constructs with pCAG-GFP. Quantification of Dab1 immunostaining in the GFP + populations in control (RVN) and (RVC + RVD) electroporated cell cultures. Mean pixel intensities of the cell soma are expressed in arbitrary units (au) after background correction (subtraction of the mean pixel intensity from a cell-free region of the same culture). A difference in mean pixel intensity is observed between RVN- and RVC + RVD-treated neuronal cultures ( p = 0.001, unpaired t test; df = 58; n = 30 GFP + cells in each condition). Neurons derived from three electroporated and pooled brains were quantified in each condition in C . Scale bar, 230 𝛍 m.
Figure 3 Suppression of Dab1 protein expression by RNAi directed against Dab1 mRNA. A , HEK293 cells cotransfected with pSuper.retro.neo plasmids directed against Dab1 (RVC + RVD) and with a plasmid encoding Dab1::GFP show reduced the amount of Dab1::GFPexpressioncomparedwithcontroltransfection(RVNwithDab1::GFP).Aplasmidencodingtheredfluorescentprotein DsRed (pDsRed) was included in all transfections as a transfection efficiency control. Three independent transfections of HEK293 cell cultures were performed to test Dab1::GFP suppression by RVN, RVC, and RVD. Green to red fluorescence ratios within regions of interest was 1.05 ± 0.25 for RVN-treated cultures ( n = 3) compared with 0.20 ± 0.23 for RVC + RVD-treated cultures ( n = 3) (means ± SD; unpaired t test, p = 0.012; df = 4). B , Western blot analysis of whole-cell lysates from HEK293 cell transfections confirms Dab1::GFP suppression after probing with anti-Dab1 antisera (B3) or with an anti-GFP antibody. Reprobing the immunoblot with an anti𝛂 -tubulin antibody confirms approximately equal loading of whole-cell lysates into each lane. C , Suppression of endogenous Dab1 in primary cortical cultures after electroporation of RNAi constructs with pCAG-GFP. Quantification of Dab1 immunostaining in the GFP + populations in control (RVN) and (RVC + RVD) electroporated cell cultures. Mean pixel intensities of the cell soma are expressed in arbitrary units (au) after background correction (subtraction of the mean pixel intensity from a cell-free region of the same culture). A difference in mean pixel intensity is observed between RVN- and RVC + RVD-treated neuronal cultures ( p = 0.001, unpaired t test; df = 58; n = 30 GFP + cells in each condition). Neurons derived from three electroporated and pooled brains were quantified in each condition in C . Scale bar, 230 𝛍 m.CC-BY-NC-SA · From the PDF
Figure 4 Dispersed cellular layering after treatment with RNAi directed against Dab1. Sections stained with the nucleic acid dye propidium iodide (red) from embryos electroporated on E16 and examined on E20 or E21 in the cingulate ( A ) or lateral neocortex ( B ). RVN control electroporated brains show precise layering by E20 and E21. B , Embryos electroporated with RNAi plasmids (RVC + RVD) directed against Dab1 show more cellular dispersion and fewer cells at the CP-MZ boundary. Scale bars: low-magnification images, 250 𝛍 m;high-magnification images, 50 𝛍 m. C , D , Binned histogram of the distribution of GFP + cell bodies in the upper 320 𝛍 mofthecingulate cortex ( C ) and lateral neocortex ( D ). Control RVN electroporations (black bars) show more cells in the upper bins (bins 1-2) than RVC + RVD (white bars). Coelectroporation of a cDNA encoding full-length Dab1 (cross-hatched bars) partially rescues RVC + RVD malpositioning. A total of 48 electroporated brains were examined, six to nine brains for each condition at each time point in the lateral neocortex, two to four brains for each condition at each time point in the cingulate cortex, and four brains for the Dab1 rescue in the lateral neocortex at E21. The soma positions for a total of 2792 cells or an average of 58 cells per brain were measured. CTX, Cortex.
Figure 4 Dispersed cellular layering after treatment with RNAi directed against Dab1. Sections stained with the nucleic acid dye propidium iodide (red) from embryos electroporated on E16 and examined on E20 or E21 in the cingulate ( A ) or lateral neocortex ( B ). RVN control electroporated brains show precise layering by E20 and E21. B , Embryos electroporated with RNAi plasmids (RVC + RVD) directed against Dab1 show more cellular dispersion and fewer cells at the CP-MZ boundary. Scale bars: low-magnification images, 250 𝛍 m;high-magnification images, 50 𝛍 m. C , D , Binned histogram of the distribution of GFP + cell bodies in the upper 320 𝛍 mofthecingulate cortex ( C ) and lateral neocortex ( D ). Control RVN electroporations (black bars) show more cells in the upper bins (bins 1-2) than RVC + RVD (white bars). Coelectroporation of a cDNA encoding full-length Dab1 (cross-hatched bars) partially rescues RVC + RVD malpositioning. A total of 48 electroporated brains were examined, six to nine brains for each condition at each time point in the lateral neocortex, two to four brains for each condition at each time point in the cingulate cortex, and four brains for the Dab1 rescue in the lateral neocortex at E21. The soma positions for a total of 2792 cells or an average of 58 cells per brain were measured. CTX, Cortex.CC-BY-NC-SA · From the PDF
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CC-BY-NC-SA · From the PDF

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Research works printed but not linked: 40

  1. (2001), J Neurophysiol, 86, 1037, 10.1152/jn.2001.86.2.1037 · DOI
  2. 10.1016/S0896-6273(00)80860-0 · DOI · Paper page
  3. 10.1002/cne.1375 · DOI
  4. 10.1016/S0896-6273(00)00007-6 · DOI · Paper page
  5. 10.1016/S0896-6273(00)80778-3 · DOI
  6. 10.1007/BF00304978 · DOI
  7. (1984), Brain Res, 318, 263
  8. 10.1523/JNEUROSCI.17-23-09204.1997 · DOI
  9. 10.1523/JNEUROSCI.21-22-08798.2001 · DOI
  10. 10.1242/dev.00654 · DOI · Paper page
  11. 10.1523/JNEUROSCI.05-12-03310.1985 · DOI
  12. 10.1038/79246 · DOI
  13. 10.1093/emboj/16.1.121 · DOI · Paper page
  14. 10.1038/39607 · DOI · Paper page
  15. 10.1101/gad.13.6.643 · DOI · Paper page
  16. 10.1002/(SICI)1097-4695(199710)33:4<459::AID-NEU9>3.0.CO;2-9 · DOI
  17. 10.1016/S0165-3806(02)00604-1 · DOI
  18. 10.1016/S0896-6273(02)00582-2 · DOI
  19. 10.1007/BF00315920 · DOI
  20. 10.1002/cne.902990107 · DOI
  21. 10.1073/pnas.2235688100 · DOI
  22. 10.1126/science.1067132 · DOI
  23. 10.1016/0165-3806(88)90121-6 · DOI
  24. 10.1038/83967 · DOI
  25. 10.1038/ng0492-50 · DOI
  26. 10.1002/cne.20322 · DOI
  27. 10.1016/S0896-6273(03)00819-5 · DOI · Paper page
  28. 10.1126/science.1411527 · DOI
  29. 10.1002/cne.901870104 · DOI
  30. (1982), Brain Res, 256, 379
  31. Ramón y Cajal S (1995) Histology of the nervous system of man and vertebrates. Vol II: Translated by N. Swanson and L. W. Swanson, Oxford: Oxford UP. · DOI
  32. 10.1523/JNEUROSCI.15-02-00981.1995 · DOI
  33. 10.1016/S0896-6273(04)00222-3 · DOI · Paper page
  34. 10.1093/cercor/10.6.602 · DOI
  35. 10.1007/s003359900240 · DOI
  36. 10.1016/S0306-4522(01)00016-1 · DOI
  37. 10.1002/jnr.10345 · DOI
  38. 10.1523/JNEUROSCI.19-23-10357.1999 · DOI
  39. 10.1016/S0092-8674(00)80782-5 · DOI · Paper page
  40. 10.1016/S0896-6273(00)80936-8 · DOI

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Metadata

Authors
Olson, E. C., Kim, S., & Walsh, C. A.
Venue
The Journal of Neuroscience
Publisher
Society for Neuroscience
Published
2006-02-08
Volume
26
Issue
6
Pages
1767-1775
ISSN
  • 0270-6474
  • 1529-2401
DOI
10.1523/jneurosci.3000-05.2006
Cited by
123
References
5
Language
en
Licence
creativecommons.org/licenses/by-nc-sa/4.0/ (vor, 181 d)

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