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. 2008 Oct 7;6(10):e246. doi: 10.1371/journal.pbio.0060246

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© 2008 Farioli-Vecchioli et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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New neurons derived from dentate gyrus progenitor cells whose differentiation was anticipated by the PC3 transgene do not become integrated into dentate gyrus spatial learning circuits.

(A) Experimental design. 2, 4, and 6 wk after treatment with BrdU, TgPC3 ON/OFF and WT mice underwent Morris water maze behavioral training, which is expected to activate new neurons.

(B) Representative confocal images of c-fos⁺ (blue), Brdu⁺ (red), NeuN⁺ (green; merged with c-fos⁺ and Brdu⁺) and of c-fos⁺/BrdU⁺/NeuN⁺ cells (white arrowhead) in the dentate gyrus following behavioral training in the 6-wk groups of TgPC3 ON and TgPC3 OFF mice (scale bar, 58 μm).

(C) New activated neurons, identified by positivity to c-fos (c-fos⁺/BrdU⁺/NeuN⁺ cells), were quantified as percentage ratio to the number of new neurons (Brdu⁺/NeuN⁺). The fraction of new neurons activated by behavioral training was null in TgPC3 OFF mice undergoing behavioral training 2 weeks after treatment with BrdU (n = 3) but became significantly detectable 4 and 6 wk after BrdU treatment (n = 6 at both time points; F(5, 274) = 7.4; p < 0.0001, ANOVA. *p < 0.05 or **p < 0.02 versus the other groups, ANOVA Fisher's PLSD post-hoc analysis). By contrast, no activation of new neurons was observed in the 2-, 4-, and 6-wk groups of TgPC3 ON mice (n = 3, 6, and 6, respectively).

(D) New neurons were generated with equivalent frequency in the TgPC3 ON and TgPC3 OFF groups, as indicated by the similar percentage ratio between number of new neurons (BrdU⁺/NeuN⁺) and total number of neurons (NeuN⁺ cells) identified 2, 4, and 6 wk after BrdU treatment (F(5, 122) = 0.24; p > 0.05, ANOVA).

(E) After behavioral training, the presence of activated neurons in the whole neuronal population of the dentate gyrus—measured as percentage ratio of c-fos⁺/NeuN⁺ to NeuN⁺ cells—was similar in all groups of TgPC3 OFF mice, whereas in TgPC3 ON mice the presence of activated neurons was significantly reduced in the 4- and 6-wk groups, indicating a progressive decrease in the population of active neurons (F(5, 112) = 16.0; p < 0.0001, ANOVA. *p < 0.02 versus 2-, 4-, 6-wk TgPC3 OFF and 2-wk TgPC3 ON mice groups, ANOVA Fisher's PLSD post-hoc analysis). The percentage ratios shown in (C–E) were calculated from absolute cell numbers measured as described in Materials and Methods and are represented as mean ± SEM.