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Table 1.

Assessment of the frequency of P/P divisions

Age
E13E14E13+E14E15
In vivo*     
   Ki67-positive/Nissl-positive 90% 83% 87% 56% 
Surface divisions in slices     
   Short culture     
      Total two-cell clones 15 19 
      Ki67-positive:Ki67-positive 11 (73%) 3 (75%) 14 (74%) 2 (33%) 
      Ki67-positive:Ki67-negative 4 (27%) 1 (25%) 5 (26%) 2 (33%) 
      Ki67-negative:Ki67-negative 0 (0%) 0 (0%) 0 (0%) 2 (33%) 
Extended culture     
   Total clones (2∼4 cells)   70  
      P/P   57 (81%)  
      P/N   11 (16%)  
      N/N   2 (3%)  
Age
E13E14E13+E14E15
In vivo*     
   Ki67-positive/Nissl-positive 90% 83% 87% 56% 
Surface divisions in slices     
   Short culture     
      Total two-cell clones 15 19 
      Ki67-positive:Ki67-positive 11 (73%) 3 (75%) 14 (74%) 2 (33%) 
      Ki67-positive:Ki67-negative 4 (27%) 1 (25%) 5 (26%) 2 (33%) 
      Ki67-negative:Ki67-negative 0 (0%) 0 (0%) 0 (0%) 2 (33%) 
Extended culture     
   Total clones (2∼4 cells)   70  
      P/P   57 (81%)  
      P/N   11 (16%)  
      N/N   2 (3%)  

Daughter cells that did not divide beyond the time that many other daughter cells had divided were judged as neurons (N) in the extended culture.

*

Fluorescent Nissl stain was used to visualize all cells in the cerebral wall. The proportion of Nissl-positive cells in the VZ and SVZ of the dorsal pallial region (photographed using a 40× objective lens) that were Ki67 positive (Fig. 1B) was calculated (n=546 cells at E13, 612 cells at E14, and 681 cells at E15)

Daughter cells generated at the ventricular surface were characterized by staining with anti-Ki67 within 4-7 hours from their birth (short culture, Fig. 1C) or by keeping them much longer (extended culture, Fig. 1D,E)

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