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Editor's Introduction

Hippocampal neurogenesis regulates forgetting during adulthood and infancy

annotated by
Sarah Moore

Why do we have so much trouble remembering our own infancy? This phenomenon, known as infantile amnesia, occurs in humans as well as many animal species. Does the synthesis of new neurons (neurogenesis), especially as our brain is developing, affect memory formation? The rate of neurogenesis is significantly higher in infancy than later in life. Is this increase related to our difficulties in forming memories during the same time period? By manipulating the rate of neurogenesis in several animal models, scientists may now have an answer to the question of forgetfulness. 

Paper Details

Original title
Hippocampal neurogenesis regulates forgetting during adulthood and infancy
Authors
Katherine G. Akers Paul W. Frankland
Original publication date
Reference
Vol. 344 no. 6184 pp. 598-602
Issue name
Science
DOI
10.1126/science.1248903

Abstract

Throughout life, new neurons are continuously added to the dentate gyrus. As this continuous addition remodels hippocampal circuits, computational models predict that neurogenesis leads to degradation or forgetting of established memories. Consistent with this, increasing neurogenesis after the formation of a memory was sufficient to induce forgetting in adult mice. By contrast, during infancy, when hippocampal neurogenesis levels are high and freshly generated memories tend to be rapidly forgotten (infantile amnesia), decreasing neurogenesis after memory formation mitigated forgetting. In precocial species, including guinea pigs and degus, most granule cells are generated prenatally. Consistent with reduced levels of postnatal hippocampal neurogenesis, infant guinea pigs and degus did not exhibit forgetting. However, increasing neurogenesis after memory formation induced infantile amnesia in these species.

Report

In both artificial systems and brain networks there is a trade-off between plasticity—the ability to incorporate new information—and stability—ensuring that the process of incorporating new information does not degrade information already stored in that network (1). In the hippocampus, new neurons continue to be generated in the subgranular zone of the dentate gyrus (DG) beyond development and into adulthood (23). These new neurons synaptically integrate into hippocampal circuits (49) and provide potential substrates for new learning. Promoting the production of new neurons in adult mice facilitates the formation of new hippocampal memories (1011). However, the continuous integration of new neurons may affect memories already stored in these circuits (12). As new neurons integrate into the hippocampus, they compete with existing cells for inputs and outputs, establishing new synaptic connections that may coexist with, or even replace, older synaptic connections (56,13). As such remodeling necessarily alters the configuration of DG-CA3 circuits and likely rescales synaptic weights of preexisting connections, computational models predict that high levels of hippocampal neurogenesis will lead to forgetting of information already stored in those circuits (1416).

Although hippocampal neurogenesis persists throughout life, rates decline dramatically with age (1718). Therefore, this predicted remodeling-induced forgetting should be most pronounced during infancy, when hippocampal neurogenesis is high. Consistent with this, infantile forgetting [or infantile amnesia (19)] is observed across a wide range of species (20), including humans (21). Neurobiological accounts of infantile amnesia previously emphasized that continued brain maturation might interfere with consolidation and/or storage of infant memories, rendering them inaccessible at later time points (22). Here, we test whether postnatal hippocampal neurogenesis, in particular, modulates ontogenetic changes in memory persistence.

Inverse Relationship Between Levels of Postnatal Neurogenesis and Memory Persistence

We first characterized levels of hippocampal neurogenesis in infant and adult mice using a retrovirus expressing green fluorescent protein (GFP) to label neural progenitors and their progeny (11) and immunohistochemistry. Four weeks after retroviral microinjections in infant (postnatal day 17; P17) and adult (P60) mice, we observed a pronounced age-dependent reduction in neurogenesis (i.e., reduction in the number of GFP+ DG granule cells and their terminal processes in the CA3 region) (Fig. 1, A and C). From infancy to adulthood, there was also a decrease in the number of proliferating (Ki67+) cells (Fig. 1D and fig. S1) and immature (doublecortin+ [DCX+]) neurons (Fig. 1, B and E). Postnatally generated DG granule cells project a mossy fiber that reaches the CA3 region after ~2 weeks, contacting 11 to 15 pyramidal cells (5723). We observed a reduction in the number of immature (DCX+) large mossy fiber terminals (LMTs) in the CA3 region of adult mice (Fig. 1, B and F), consistent with a predicted reduction in synaptic rearrangements in the CA3 with age (fig. S2).

figure 1

Fig. 1.  Age-dependent levels of hippocampal neurogenesis and memory stability are inversely related in mice. (A) GFP labeling of granule cells in the DG and CA3 1 month after retroviral infection shows that infant (P17) mice had higher levels of neurogenesis than adult (P60) mice. (Left) Low magnification of DG and CA3 regions. Scale bar, 200 μm. (Right) High magnification of CA3 region showing GFP+ mossy fibers and LMTs. Scale bar, 50 μm. (B) Top row: Infants had more DCX+ cells in the DG and CA3 than adults. Scale bar, 100 μm. Bottom row: High magnification of DG and CA3. Scale bar, 50 μm. (C to F) Infants showed more (C) GFP+ neurons (t6 = 8.57, P< 0.001), (D) proliferating (Ki67+) cells (t7 = 4.53, P < 0.01), and (E) immature (DCX+) neurons in the DG (t6 = 18.65, P < 0.001), as well as more (F) DCX+ LMTs in the CA3 (t6 = 15.03, P < 0.001) than adults (n = 4 to 5 per group). (G) Separate groups of infant and adult mice were trained in contextual fear conditioning and tested 1, 7, 14, or 28 days later (n = 8 to 9 per group). (H) Context fear memory persisted for at least 28 days in adult mice, whereas infants showed intact memory shortly after training but forgetting after longer delays (age × dayF3,62 = 13.07, P < 0.001; post-hoc t tests, 1 day, P < 0.05; 7 days, P < 0.001; 14 days, P < 0.001; 28 days, P < 0.001). (I) Infants and adults showed equivalent shock reactivity during training (P17 n = 9, P60 n = 8; arbitrary units, P > 0.05). (J) Inverse relationship between neurogenesis and memory persistence predicts that increasing neurogenesis in adults will induce forgetting, whereas decreasing neurogenesis in infants will mitigate forgetting. In all panels, cells/LMTs are expressed as number of cells/LMTs per 1000 μm2. *P < 0.05. For all figures, error bars represent standard error of the mean (SEM). DAPI, 4′,6-diamidino-2-phenylindole.

Hypothesis

The rate of neurogenesis decreases with age while memory stability increases with age.

Experiments

Retroviral-GFP labeling to compare the amount of neurogenesis in infant and adult mice.

Immunohistochemistry for DCX and Ki67 to compare the amount of neurogenesis in infant and adult mice.

Context fear conditioning memory test to compare memory stability in infant and adult mice.

Conclusions

There are more GFP+ cells in infant than in adult mice, indicating that the amount of neurogenesis is higher in infant mice than adults.

There are more DCX+ cells and LMTs in infant than in adult mice, indicating that there are more immature neurons (with immature synapses) in infant than in adult mice. This is further evidence that the rate of neurogenesis is higher in infant mice.

There are more Ki67+ cells in infant than in adult mice, indicating that there are more actively dividing cells. This also supports the conclusion that the amount of neurogenesis decreases with age.

Context fear memory was maintained for the duration of the study (28 days) in adult mice, but was lost in infant mice after only 1 day. However, infants and adults both respond to the initial fear conditioning the same way (as measured by shock reactivity).

Significance

This figure confirms that there is an inverse relationship between neurogenesis and memory stability. 

Next, we evaluated how age-dependent changes in neurogenesis influence the ability to form enduring hippocampus-dependent memories using the contextual fear conditioning paradigm (24). Infant and adult mice were placed in a novel context and presented with a series of foot shocks. One to 28 days later, separate groups of mice were returned to the context, and memory retention was evaluated by measuring freezing behavior (Fig. 1G). Mice trained as adults froze robustly regardless of retention delay. By contrast, mice trained as infants only exhibited high levels of freezing when tested 1 day after training. If infant mice were tested at longer retention delays, less freezing was observed (Fig. 1H) (25). Infant mice trained similarly but not shocked showed little freezing when tested 1 day after training (fig. S3), indicating that the freezing observed in shocked infant mice reflects conditioned fear. There were no age-related differences in reaction to the foot shock (Fig. 1I), indicating that forgetting in infant mice was not due to diminished nociception. This pattern of accelerated forgetting in infant mice was replicated using an incidental context learning paradigm (fig. S4).

Increasing Neurogenesis Promotes Forgetting in Adult Mice

To determine if levels of neurogenesis and memory persistence are causally related, we tested whether increasing hippocampal neurogenesis after learning promotes forgetting in adult mice (Fig. 1J) (26). We initially used voluntary running, a naturalistic intervention that robustly increases neurogenesis (27) (Fig. 2A). Relative to sedentary controls, running increased the number of proliferating (Ki67+) cells (Fig. 2B) and immature (DCX+) neurons in the DG (Fig. 2C), as well as DCX+ LMTs in the CA3 of adult mice (Fig. 2D). These increases were evident within 7 days of commencing running, sustained for the duration of exercise, and not associated with increased death of developmentally generated granule cells (fig. S5). We next injected retrovirus expressing GFP into the DG of adult mice. Four weeks later, we observed many GFP+ neurons in the innermost layers of the DG and GFP+ LMTs in the stratum lucidum of the CA3. Counterstaining with a marker of mature presynaptic terminals, zinc transporter-3 (ZnT3) (28), revealed many new (GFP+/ZnT3+) LMTs in close apposition to preexisting (GFP–/ZnT3+) LMTs (Fig. 2E), suggesting that new contacts coexist with established connections (5). Running increased the ratio of new-to-established LMTs, indicating that running increases remodeling of DG-CA3 circuits (fig. S6).

figure 2

Fig. 2.  Voluntary running increases neurogenesis and promotes forgetting in adult mice. (A) Adult (P60) mice given access to a running wheel for 28 days showed more immature (DCX+) neurons in the DG and LMTs in the CA3 than sedentary mice. Scale bar, 50 μm. (B to D) Adult mice were given access to a running wheel for 0, 3, 7, 14, or 28 days. Running increased the number of (B) proliferating (Ki67+) cells (n = 8 per group; running × dayF4,70 = 7.71, P < 0.001; post hoc t tests, 7 days, P < 0.001; 14 days, P < 0.001; 28 days,P = 0.001), and (C) immature (DCX+) neurons in the DG (n = 4 to 8 per group; running × dayF4,34 = 3.17, P < 0.05; 7 days, P < 0.05; 14 days, P < 0.001; 28 days, P < 0.05), as well as more (D) DCX+ LMTs in the CA3 (n = 4 per group; running × dayF4,30 = 3.45, P < 0.05; 7 days, P= 0.001; 14 days, P < 0.05; 28 days, P < 0.05) compared with sedentary controls. (E) Neurogenesis remodels DG-CA3 circuits. Retrovirus-labeled presynaptic LMTs from new neurons (GFP+) in close apposition to existing presynaptic LMTs from mature neurons (ZnT3+). Scale bar, 50 μm. Confocal stacks used to reconstruct representative three-dimensional images of new LMTs (green) in close contact with existing LMTs (red) in the CA3. (F) Adult mice ran or remained sedentary after contextual fear conditioning. (G and H) Running increased neurogenesis (sedentary n = 8, running n = 8; LacZ+ cells; t14 = 2.81, P < 0.05) and induced forgetting (sedentary n = 9, running n = 8; t15 = 2.75, P < 0.05). (I) Adult mice ran or remained sedentary before contextual fear conditioning. (J and K) Running increased neurogenesis (sedentary n = 7, running n = 8; t13 = 2.96, P < 0.05) but did not affect formation of a context fear memory (sedentary n = 10, running n = 9; P > 0.05). In all panels, cells/LMTs are expressed as number of cells/LMTs per 1000 μm2. *P < 0.05.

Hypothesis

Increasing neurogenesis is sufficient to decrease memory stability in adult mice.

Experiments

Immunohistochemistry (as in Figure 1) to compare the amount of neurogenesis in adult mice with and without a running wheel in their cage.

Retroviral-GFP labeling and Znt3+ immunohistochemistry to determine if new and old synapses localize together.

Context fear conditioning memory test to compare memory stability in running versus sedentary mice. In separate experiments, mice were provided with a running wheel (or not) before fear conditioning or after fear conditioning to determine the effect of running on successful fear conditioning versus maintaining a memory.

Conclusions

There are more DCX+ neurons and LMTs and Ki67+ cells in mice provided with a running wheel than in sedentary controls. This indicates that running increases neurogenesis.

GFP+/Znt3+ new synapses are in close apposition with GFP+/Znt3+ old synapses. This indicates that the incorporation of new synapses may disrupt the stability of old synapses and cause DG-CA3 remodeling.

Running decreased memory persistence in adult mice.

Running did not affect the acquisition of a contextual fear memory.

Significance

This figure demonstrates that increasing neurogenesis in adult mice is sufficient to induce DG-CA3 remodeling and decrease memory stability.

To determine whether running-induced increases in hippocampal remodeling induce forgetting of an established hippocampus-dependent memory, we trained adult mice in contextual fear conditioning. After training, mice were given continuous access to a running wheel in their home cage or housed conventionally (i.e., sedentary; Fig. 2F). Running increased neurogenesis in the DG (Fig. 2G) and reduced context fear memory when mice were tested 6 weeks later (Fig. 2H and fig. S7). Access to a locked running wheel did not induce forgetting (fig. S8), indicating that forgetting was associated with running rather than exposure to a novel object in the home cage. Furthermore, running-induced forgetting of a context fear memory was reduced after a stronger training protocol (i.e., additional foot shocks; fig. S9), suggesting that the degree of forgetting depends, in part, on initial memory strength.

As voluntary running induces several neural and physiological changes apart from increasing hippocampal neurogenesis (29), forgetting might alternatively be mediated by these non-neurogenic changes. Therefore, we first examined the impact of running before (rather than after) training (Fig. 2I). Running before training similarly increased neurogenesis (Fig. 2J) but did not affect acquisition of a context fear memory (Fig. 2K), indicating that running does not nonspecifically alter freezing (e.g., by reducing anxiety).

Second, we tested whether preventing the running-induced increase in neurogenesis would prevent forgetting using transgenic mice in which a nestin promoter/enhancer drives expression of a modified herpes simplex virus (HSV) gene encoding thymidine kinase (HSV-tk) (TK+ mice) (30). In TK+ mice, administration of ganciclovir (GAN) ablates only dividing cells expressing the tk transgene. TK+ mice and their wild-type (WT) littermates were trained in contextual fear conditioning and then given access to a running wheel or housed conventionally for 6 weeks. During this period, mice were treated with GAN to suppress neurogenesis (Fig. 3A). GAN treatment in TK+ mice limited the running-induced increase in neurogenesis to WT sedentary levels (Fig. 3B) and prevented forgetting (Fig. 3C).

figure 3

Fig. 3.  Forgetting depends on increased neurogenesis. (A) Adult GAN-treated WT and TK+ mice remained sedentary or ran after contextual fear conditioning. Running (B) increased neurogenesis (sedentary n = 6, running n = 14; t18 = 2.81, P < 0.05) and (C) induced forgetting of a context fear memory in WT mice (sedentary n = 16, running n = 14; t28 = 2.63, P < 0.05). By contrast, in TK+ mice, both (B) the running-induced increase in neurogenesis (sedentary n = 8, running n = 9; TK+ running versus WT sedentary, P > 0.05; WT running versus TK+ running, t21 = 3.43, P < 0.01) and (C) forgetting (TK+sedentary n = 12, TK+ running n = 10; P > 0.05; genotype × running interaction, F1,48 = 4.88, P < 0.05) were blunted. (D) Adult WT mice treated with MEM after contextual fear conditioning showed (E) increased neurogenesis (vehicle n = 7, MEM n = 6; t11 = 2.55, P < 0.05) and (F) forgetting of a context fear memory (vehicle n = 8, MEM n = 8; t14 = 5.47, P < 0.001) relative to vehicle-treated controls. (G) Adult GAN-treated WT and TK+ mice were treated with vehicle or MEM after contextual fear conditioning. MEM (H) increased neurogenesis (vehicle n = 8, MEM n = 8; t14 = 2.25, P < 0.05) and (I) induced forgetting of a context fear memory in WT mice (vehicle n = 11, MEM n = 11; t20 = 2.23, P < 0.05). By contrast, in TK+ mice, both (H) the MEM-induced increase in neurogenesis (vehicle n = 8, MEM n = 8; TK+ vehicle versus TK+ MEM, P > 0.05; WT MEM versus TK+ MEM, t14 = 8.53, P < 0.001; genotype × drug interaction, F1,28 = 4.69, P < 0.05) and (I) forgetting (vehicle n = 13, MEM n = 14; TK+ vehicle versus TK+ MEM, P > 0.05; genotype × drug interaction,F1,45 = 5.41, P < 0.05) were attenuated. (J) Post-training tamoxifen treatment (K) increased neurogenesis (WT n = 4, iKO-p53 n = 4; t6 = 4.39, P < 0.01) and (L) induced forgetting of a context fear memory (WT n = 14, iKO-p53 n = 9; t21 = 2.68, P < 0.05) in iKO-p53 mice relative to WT controls. In all panels, cell counts are expressed as number of cells per 1000 μm2. *P < 0.05.

Hypothesis

Increased neurogenesis is necessary for forgetting.

Experiments

Adult WT and TK+ mice were treated with GAN, which should inhibit neurogenesis in TK+ mice but not WT controls, to determine if running still decreases memory stability if the increased neurogenesis associated with running is inhibited.

Adult WT and TK+ mice were treated with MEM to determine if the same effects on increasing neurogenesis and decreasing memory stability are present with a method other than running.

Adult WT and TK+ mice were treated with MEM and GAN to determine if MEM still decreases memory stability if the increased neurogenesis associated with the drug is inhibited.

Adult iKO-p53 and WT mice were treated with Tamoxifen, which will prevent p53 expression in iKO-p53 but not WT mice, to determine if increased neurogenesis associated with p53 knockdown results in decreased memory stability.

Conclusions

GAN treatment of TK+, but not WT mice, prevented the running-induced increase in neurogenesis and the associated decrease in memory stability.

MEM treatment increased neurogenesis and decreased memory stability.

GAN treatment of TK+, but not WT mice, prevented the MEM-induced increase in neurogenesis and the associated decrease in memory stability.

Tamoxifen treatment of iKO-p53 mice, but not WT mice, increased neurogenesis and decreased memory stability.

Significance

This figure demonstrates that increased neurogenesis is necessary for the decrease in memory stability associated with both running and MEM treatment.

Third, nonrunning interventions that increase neurogenesis might similarly induce forgetting of established memories. Administering the proneurogenic drugs memantine (MEM) (31) (Fig. 3, D to F, and fig. S10) or fluoxetine (32) (fig. S11) after training induced forgetting [see also (33)]. Both the increase in neurogenesis and forgetting induced by MEM were blocked by activation of the tk transgene in TK+ mice (Fig. 3, G to I). We also generated transgenic mice in which the tumor suppressor gene p53 was inducibly deleted from neural progenitor cells and their progeny during adulthood (iKO-p53 mice) (Fig. 3J). Similar to previous studies examining global deletion of p53 (34), conditional deletion of p53 increased the number of proliferating (Ki67+) cells (fig. S12) and immature (DCX+) neurons (Fig. 3K) in the DG. Furthermore, post-training deletion of p53 induced forgetting of context fear memory in adult mice (Fig. 3L).

Finally, running also induced forgetting of other hippocampus-dependent memories (incidental context learning, fig. S13; water maze, fig. S14). However, running did not induce forgetting of a conditioned taste aversion memory (fig. S15), which does not depend on the hippocampus (35).

Reducing Neurogenesis Increases Memory Persistence in Infant Mice

Using genetic and pharmacological strategies, we next examined whether decreasing neurogenesis mitigates forgetting normally observed in infant mice (Fig. 1J). To genetically suppress neurogenesis, we administered GAN to infant TK+ and WT mice (Fig. 4A). GAN treatment reduced the number of proliferating (Ki67+) cells and immature (DCX+) neurons in the DG of TK+ mice compared with WT littermates (Fig. 4, B and C). When tested 1 day after contextual fear conditioning, WT and TK+ mice exhibited equivalent levels of freezing (Fig. 4D), indicating normal memory at this short retention delay. However, when separate groups of mice were tested 7 days after training, TK+ mice froze more than WT mice (Fig. 4E), and freezing levels were inversely correlated with levels of neurogenesis (Fig. 4F). GAN-treated WT and TK+ mice trained without foot shocks showed little freezing when tested 7 days after training (Fig. 4E). These low levels of freezing were similar to those observed in shocked WT mice, indicating substantial forgetting in WT but not TK+ mice. Furthermore, the absence of freezing in nonshocked TK+ mice suggests that suppressing neurogenesis (or GAN treatment) does not simply increase the propensity to freeze.

figure 4

Fig. 4.  Genetic and pharmacological decreases in neurogenesis mitigate forgetting in infant mice. (A) Infant GAN-treated WT or TK+ mice were trained in contextual fear conditioning or placed in conditioning chambers without foot shocks. TK+ mice had fewer (B) proliferating (Ki67+) cells (WT n = 9, TK+ n = 14; t21 = 3.08, P < 0.01) and (C) immature (DCX+) neurons (WT n = 6, TK+ n = 6; t10 = 2.57, P < 0.05) in the DG than WT mice. (D) Shocked WT and TK+ mice showed equivalent context fear memory 1 day after training (WTn = 9, TK n = 13; P > 0.05), and nonshocked mice showed low freezing (WT n = 7, TK n = 9). (E) A separate group of shocked WT mice showed forgetting of a context fear memory 7 days after training, whereas this forgetting was reduced in a separate group of TK+ mice (WT n = 9, TK+ n = 14; t21 = 2.26, P < 0.05). Separate groups of nonshocked mice again showed low freezing (WT n = 6, TK n = 7). (F) Inverse relationship between proliferation and memory persistence among shocked mice 7 days after training (r = –0.46, P < 0.05). (G) Infant mice were treated with vehicle or TMZ after training in the incidental context learning paradigm (or were similarly treated without foot shocks). TMZ-treated mice had fewer (H) proliferating (Ki67+) cells (vehicle n = 30, TMZ n = 29; t57 = 3.57, P = 0.001) and (I) immature (DCX+) neurons (vehicle n = 5, TMZ n = 6; t9 = 2.29, P < 0.05) in the DG. (J) Shocked TMZ-treated mice showed less forgetting of a context-only memory (vehicle n = 30, TMZ n = 29; t57 = 2.47, P < 0.05). Nonshocked mice showed low freezing (vehicle n = 9, TMZ n = 9). (K) Inverse relationship between proliferation and memory persistence among shocked mice (r = -0.31, P < 0.05). In all panels, cells are expressed as number of cells per 1000 μm2. *P < 0.05.

Hypothesis

The high rate of neurogenesis in infant mice is necessary for exhibition of infantile amnesia.

Experiments

Infant WT and TK+ mice were treated with GAN to genetically decrease neurogenesis in TK+ but not WT mice.  Mice were tested for context fear memory to determine if decreasing neurogenesis increases memory stability in infant mice.

Infant mice were treated with a control vehicle or TMZ drug to pharmacologically inhibit neurogenesis.  Mice were tested for context fear memory to determine if decreasing neurogenesis increases memory stability in infant mice.

Conclusions

GAN treated TK+ mice had fewer Ki67+ and DCX+ cells than WT controls, indicating that GAN treatment inhibits neurogenesis in TK+ but not WT mice.

Both WT and TK+ mice respond to context fear conditioning.

Mice that were not shocked do not develop context fear conditioning (regardless of GAN or TMZ treatment).

GAN treatment of infant TK+ mice decreases forgetting compared to WT controls.

TMZ treated mice had fewer Ki67+ and DCX+ cells than vehicle controls, indicating that TMZ treatment inhibits neurogenesis.

TMZ treatment of infant mice decreases forgetting compared to vehicle controls.

Significance

This figure demonstrates that decreasing neurogenesis in infant mice decreases infantile forgetting.

figure 5

Fig. 5.  Precocial degus and guinea pigs show low postnatal neurogenesis and persistent infantile memories. (A) DCX+ cells in the DG of infant and adult degus. (Left) Low magnification. Scale bar, 200 μm. (Right) High magnification. Scale bar, 25 μm. (B) DCX+ cells in the DG of infant and adult guinea pigs. (Left) Low magnification. Scale bar, 200 μm. (Right) High magnification. Scale bar, 25 μm. Compared with mice (P17n = 4 to 5, P60 n = 4), degus (P17 n = 3, P60 n = 4 to 6) and guinea pigs (GP; P17 n = 4, P60 n = 4) showed reduced age-related declines in (C) proliferating (Ki67+) cells and (D) immature (DCX+) neurons in the DG. (E) Infant and adult degus were trained in contextual fear conditioning and tested 1 and 28 days later (P17 n = 7, P60 n = 10). (F) Both infant and adult degus showed persistent context fear memory (age × delay interaction and agemain effect, Ps > 0.05). (G) Separate groups of infant and adult guinea pigs were trained in a water maze and tested 1 (P17 n = 8, P60 n = 8) or 28 days later (P17 n = 8, P60 n = 8). (H) Both infant and adult guinea pigs showed persistent spatial memory during probe tests (age × delay interaction, P > 0.05; age main effect, F1,28= 6.05, P < 0.05).

Hypothesis

Animals that are more developed at birth and have lower rates of neurogenesis, as infants do not exhibit infantile amnesia.

Experiments

Immunohistochemistry with Ki67+ and DCK+ in infant and adult degus and guinea pigs to compare age-related decreases in neurogenesis between degus, guinea pigs, and mice.

Contextual fear memory testing to determine if degus exhibit infantile amnesia.

A water maze was used to test the spatial memory of guinea pigs (recall that spatial memory is also dependent on the hippocampus and is included in the infantile amnesia phenomenon). 

Conclusions

Degus and guinea pigs have a smaller decrease in neurogenesis (DCX+ and Ki67+ cells) between infants and adults than mice. This indicates that the amount of neurogenesis in degus and guinea pigs is already smaller at birth than in mice.

Degus do not show decreased retention of a contextual fear memory as infants.

Guinea pigs do not show decreased retention of spatial memories as infants.

Significance

This figure demonstrates that animals with lower levels of neurogenesis as infants do not exhibit infantile amnesia.

Using temozolomide (TMZ), a DNA alkylating agent (36), to pharmacologically reduce hippocampal neurogenesis in infant WT mice, we observed a similar attenuation of forgetting in an incidental context learning paradigm. Infant mice were preexposed to a context and then treated with TMZ or vehicle for 4 weeks (Fig. 4G). TMZ treatment suppressed neurogenesis (Fig. 4, H and I) and improved retention of the context-only memory (Fig. 4J), and there was an inverse relationship between hippocampal neurogenesis and freezing (Fig. 4K). TMZ treatment did not decrease the threshold for freezing, however, as similar TMZ treatment before learning did not alter freezing levels (fig. S16). During infancy, high levels of proliferation are associated with high levels of apoptotic cell death in the subgranular zone (37). However, post-training pharmacological inhibition of cell death alone failed to attenuate forgetting (fig. S17).

Infantile Forgetting Is Absent in Precocial Guinea Pigs and Degus

As guinea pigs have a longer (~65 days) gestation than mice (~21 days), they are more neurologically mature at birth and have reduced postnatal hippocampal neurogenesis (38). This led us to predict that guinea pigs (and possibly other rodents with similarly extended gestation) will not exhibit infantile forgetting. To test this, we examined hippocampal neurogenesis and memory retention in guinea pigs [in which postnatal hippocampal neurogenesis has been described (38)] and degus, another precocial rodent (in which postnatal neurogenesis has not previously been studied). Whereas there is a considerable reduction in proliferating (Ki67+) cells and immature (DCX+) neurons from P17 to P60 in mice, this reduction was much more modest in degus and guinea pigs (Fig. 5, A to D). Unlike infant mice that showed rapid forgetting, infant degus showed normal retention of a context fear memory for up to 1 month (Fig. 5, E and F), and infant guinea pigs showed no change in spatial memory as a function of retention delay (Fig. 5, G and H), indicating that memories are persistent in infant rodent species with low postnatal hippocampal neurogenesis.

Increasing Hippocampal Neurogenesis Is Sufficient to Induce Forgetting in Infant Guinea Pigs and Degus

If levels of hippocampal neurogenesis and forgetting are causally related, then increasing hippocampal neurogenesis in degus and guinea pigs should be sufficient to induce infantile amnesia in these precocial species. To test this, infant (P17) degus were trained in contextual fear conditioning and then either housed conventionally, given access to a running wheel, or treated with MEM or vehicle for 4 weeks (Fig. 6, A and D). Both running and MEM treatment increased neurogenesis (Fig. 6, B and E) and induced forgetting (Fig. 6, C and F). Similarly, forgetting of a spatial memory was observed after MEM treatment in infant guinea pigs (Fig. 6, G to I). These results indicate that increasing hippocampal neurogenesis using mechanistically distinct interventions induces forgetting in precocial rodent species.

Discussion

The hippocampus encodes memories for places and events (39). The observation that hippocampal neurogenesis persists into adulthood led to the idea that neurogenesis modulates hippocampal memory function (40). To our knowledge, all previous studies examining the relationship between hippocampal neurogenesis and memory have used essentially the same design; they manipulated hippocampal neurogenesis before training and examined the impact of this manipulation on subsequent memory formation (i.e., they investigated the anterograde effects of manipulating neurogenesis on memory) (40). The view that emerged from these studies is that, once sufficiently mature, new neurons positively contribute to encoding of new hippocampus-dependent memories, perhaps by providing new substrates for memory storage (40). Here, we examined the retrograde impact of similar manipulations of neurogenesis on memory. Through a series of studies, we showed that high levels of neurogenesis disrupt established hippocampus-dependent memories. As such, our findings reveal a novel role for neurogenesis in forgetting or memory clearance, in line with theoretical predictions (121416).

figure 6

Fig. 6.  Increasing neurogenesis promotes forgetting in infant degus and guinea pigs. (A) Infant degus were trained in contextual fear conditioning and then remained sedentary or given access to a running wheel for 28 days. Running (B) increased neurogenesis (sedentary n = 4, running n = 4; t6 = 4.74, P < 0.01) and (C) induced forgetting of a context fear memory (sedentary n = 21, running n = 16; t35 = 4.59, P < 0.001). (D) Infant degus were trained in contextual fear conditioning and then treated with vehicle or MEM for 28 days. MEM treatment (E) increased neurogenesis (vehicle n = 8, MEMn = 5; t11 = 4.41, P = 0.001) and (F) induced forgetting of a context fear memory (vehicle n = 12, MEM n = 11; t21 = 5.35, P < 0.001). (G) Infant guinea pigs were treated with vehicle or MEM after training in the water maze. MEM treatment (H) increased neurogenesis (vehicle n = 3, MEM n = 3; t4 = 3.11, P < 0.05) and (I) induced forgetting of a spatial memory (vehicle n = 11, MEM n = 12; t21 = 3.32, P < 0.01). In all panels, cells are expressed as number of cells per 1000 μm2. *P < 0.05.

Hypothesis

Increasing neurogenesis in infant precocial species will induce infantile amnesia.

Experiments

Infant degus were given access to a running wheel (or not) following contextual fear conditioning to determine if running-induced increases in neurogenesis would promote forgetting.

Infant degus were treated with MEM or vehicle control following contextual fear conditioning to determine if MEM-induced increases in neurogenesis would promote forgetting.

Infant guinea pigs were treated with MEM or vehicle control following water maze training to determine if MEM-induced increases in neurogenesis would promote forgetting.

Conclusions

Running increases DCX+ cells and induces forgetting in infant degus.

MEM increases DCX+ cells and induces forgetting in infant degus and guinea pigs.

Significance

This figure demonstrates that increasing neurogenesis in precocial species is sufficient to cause infantile forgetting.

The hippocampus is thought to rapidly and automatically encode experiences (41). Because not all experiences are ultimately remembered, it is likely that forgetting processes continuously degrade or clear stored information from the hippocampus. Our results identify neurogenesis as one such process that promotes degradation of hippocampus-dependent memories, most likely by reconfiguring DG-CA3 circuits. Successful memory retrieval may result from the reactivation of patterns of neural activity present at the time of memory encoding (i.e., pattern completion) [e.g., (4245)]. Because neurogenesis reconfigures hippocampal circuits, this may reduce the ability of a given set of cues (or inputs) to reinvoke the same pattern of activity (i.e., pattern completion failure) (46). During infancy, when neurogenesis levels are elevated, high rates of decay render hippocampus-dependent memories [that are declarative in nature (21)] inaccessible at later time points. Reducing neurogenesis at this developmental stage can increase the persistence of hippocampus-dependent memories. During adulthood, when neurogenesis levels are lower, memories are more resistant to decay. Artificially increasing neurogenesis after learning, however, may be sufficient to induce forgetting.

Supplementary Materials

www.sciencemag.org/content/344/6184/598/suppl/DC1

Materials and Methods

Figs. S1 to S17

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Acknowledgments: This work was supported by grants from Brain Canada to P.W.F., the Canadian Institutes of Health Research (CIHR) to P.W.F. (MOP86762) and S.A.J. (MOP74650), the Centre for Brain and Behavior at the Hospital for Sick Children to P.W.F. and S.A.J., and the Core Research for Evolutional Science and Technology of Japan Science and Technology Agency (JST-CREST) and Ministry of Education, Culture, Sports, Science and Technology Grants-in-Aid for Scientific Research (KAKENHI) to T.M. K.G.A. and H.-L.H. (CIHR), A.M.-C. (El Consejo Nacional de Ciencia y Tecnología), A.P.Y. (Donald and Doris Milne Alzheimer’s Disease Fellowship), A.L.W. (Ontario Mental Health Foundation), and A.G. and B.A.R. (Natural Sciences and Engineering Research Council of Canada) were supported by fellowships. Data are archived in the Dryad repository (doi:10.5061/dryad.s7q3c).