Gretchen D Frantz, Susan K McConnell  Neuron 

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Restriction of Late Cerebral Cortical Progenitors to an Upper-Layer Fate  Gretchen D Frantz, Susan K McConnell  Neuron  Volume 17, Issue 1, Pages 55-61 (July 1996) DOI: 10.1016/S0896-6273(00)80280-9

Figure 1 Positions of Isochronically and Heterochronically Transplanted Cells in Host Brains (A) DiI-labeled, isochronically transplanted cells (arrows) in the cortical ventricular zone (vz) of an E32 ferret immediately after transplantation. (B and C) Positions of [3H]thymidine-labeled cells in the developing telencephalon before the onset of cortical upper-layer neurogenesis. (B) Transplanted cells (arrow) with an elongated morphology in the cortical plate following isochronic transplantation (8 day survival). (C) Labeled cell (arrow) with an elongated, migratory morphology at the top of the cortical plate after heterochronic transplantation (9 day survival). Because most labeled cells will eventually populate the upper layers, the neurons of which are only beginning to be generated in the host, it is likely that this transplanted cell will continue to migrate outward as new cells are added to the top of the cortical plate. (D and E) Final positions of differentiated [3H]thymidine-labeled transplanted cells in the adult cortex. (D) Heavily labeled pyramidal neuron (arrow) and lightly labeled neuron (arrowhead) in layer 5 of an isochronic transplant recipient. (E) Heavily labeled neuron (arrow) in layer 2/3 of a heterochronic transplant recipient. Abbreviations: iz, intermediate zone; mz, marginal zone. Scale bar in (E) represents 200 μm for (A) and 20 μm for (B)–(E). Neuron 1996 17, 55-61DOI: (10.1016/S0896-6273(00)80280-9)

Figure 2 Migration of Transplanted Neurons in the Host Cerebral Wall Low power photomicrographs of an isochronic host animal, 8 days after transplantation. (A) Dark-field view in which [3H]thymidine-labeled cells are visible as white dots. Many labeled cells have entered the cortical plate (cp), where they accumulate underneath the marginal zone (mz). Other labeled cells are found in the intermediate zone (iz) and in an injection site in the ventricular zone (vz). (B) Bright-field view of the same section, stained with cresyl violet. The injection site is visible as a small bulge at the lumenal surface (asterisk). Scale bar represents 300 μm. Neuron 1996 17, 55-61DOI: (10.1016/S0896-6273(00)80280-9)

Figure 3 Final Laminar Positions of [3H]Thymidine-Labeled, Transplanted Cells That Have Migrated into the Adult Host Cortex Closed bars show the positions of cells heavily labeled with [3H]thymidine (20 silver grains/cell), and open bars show the positions of lightly labeled cells (<20 grains/cell). (A) Isochronic transplants: heavily labeled cells (n = 310 cellsfrom 3 brains) are found predominantly in layer 5, while lightly labeled cells (n = 367) populate slightly more superficial positions. There was no statistically significant difference between these two distributions by χ2 analysis (p = 0.858). (B) Heterochronic transplants: both heavily labeled (n = 590 cellsfrom 10 brains) and lightly labeled cells (n = 139) are found almost exclusively in layer 2/3, the position appropriate for their origin. The distribution of each of these populations differs significantly from those of isochronic controls (χ2 analysisp = 0.0001), but not from one another (p = 0.679). Abbreviations: wm, white matter; uwm, former subplate cells in the white matter directly underlying layer 6. Neuron 1996 17, 55-61DOI: (10.1016/S0896-6273(00)80280-9)

Figure 4 Possible Mechanisms for the Targeted Migration of Upper-Layer Cells Following Transplantation into a Younger Host Brain Each panel shows a successively later time in development. One possibility (stippled cells) is that transplanted cells remain in the ventricular zone until the onset of upper-layer neurogenesis in the host, and only then migrate. Alternatively (solid cells), cells might initiate migration immediately after transplantation, but continue to migrate until upper-layer host neurons arrive in the cortical plate. Abbreviations: mz, marginal zone; cp, cortical plate; iz, intermediate zone; vz, ventricular zone. Neuron 1996 17, 55-61DOI: (10.1016/S0896-6273(00)80280-9)

Figure 5 Positions of Migrating [3H]Thymidine-Labeled Cells at Various Times after Transplantation (A) and (B) show times that precede the onset of upper-layer neurogenesis, while (C) and (D) show times at which deep-layer neurons are differentiating and upper-layer neurons are just beginning to enter the cortical plate. Cells remaining at injection sites are not included (see Experimental Procedures). (A) Isochronic transplants, 7–8 day survival: 86% (n = 419 cells, 2 brains) of migrating transplanted cells have entered the cortical plate. (B) Heterochronic transplants, 8–9 day survival: 50% (n = 507 cells, 3 brains) of migrating cells are in the cortical plate. This distribution differs significantly from that of the comparably aged isochronic controls shown in (A) (χ2 analysisp = 0.0001). (C) Isochronic transplants, 12 day survival: roughly 72% (n = 395 cells, 4 brains) of cells that have migrated away from injection sites are in their final positions within the lower two-thirds of the cortical plate. (B) Heterochronic transplants, 16 day survival: 87% (n = 470 cells, 1 brain) of migrating transplanted cells are in the cortical plate; the majority occupy its upper zone. This distribution differs significantly from that shown in (C) (χ2 analysisp = 0.0001). Abbreviations: MZ, marginal zone; CPu, upper portion of the cortical plate, containing newly arrived neurons; CPm, middle portion of the cortical plate; CPl, lower portion of the cortical plate; IZ, intermediate zone. Neuron 1996 17, 55-61DOI: (10.1016/S0896-6273(00)80280-9)