Develop. Growth Differ. (2009) 51, 379–386 doi: 10.1111/j.1440-169X.2009.01094.

Review
Blackwell Publishing Asia

Continuous neurogenesis in the adult brain

Itaru Imayoshi1,2*, Masayuki Sakamoto1,3, Toshiyuki Ohtsuka1,2 and
Ryoichiro Kageyama1,2
1
Institute for Virus Research, Kyoto University, Kyoto 606-8507, 2Japan Science and Technology Agency, CREST,
Kyoto 606-8507, and 3Kyoto University Graduate School of Biostudies, Kyoto 606-8502, Japan

Neurogenesis occurs throughout adulthood in the mammalian brain by coordinated proliferation and differentiation
of adult neural stem cells. Newborn neurons are incorporated into the functional networks of both the olfactory
bulb and the hippocampal dentate gyrus, suggesting significant roles of adult neurogenesis in brain functions. In
this review, we discuss the recent findings about the integration mode of new neurons into the existing neural
circuits. We further address the potential significance of adult neurogenesis in higher brain functions such as
olfactory and spatial memory.

Key words: adult neurogenesis, Cre, dentate gyrus, neural stem cell, olfactory bulb.

(SVZ) of the lateral ventricles and the subgranular zone

Introduction
It is now widely accepted that neurogenesis occurs
continuously in the forebrain of adult mammals, including
humans (McKay 1997; Gage 2000; Alvarez-Buylla et al.
2001; Temple 2001; Doetsch 2003; Ming & Song 2005).
The generation of new neurons in the adult brain was
first reported by Altman and colleagues using the [H3]-
thymidine-incorporation labeling method in the dentate
gyrus of the rat hippocampus (Altman & Das 1965).
They reported a series of papers about the neurogenesis
in various regions of adult rats, including neocortex and
olfactory bulb (Altman 1966, 1969). Long-term survival
of newborn neurons in the hippocampus was also
demonstrated (Kaplan & Hinds 1977). The development
of a bromodeoxyuridine (BrdU)-incorporation labeling
method enabled us to analyze the characteristics of
newborn neurons by combination with immunohisto-
chemistry. Adult neurogenesis was observed with
BrdU incorporation in all mammals examined, including
humans (Eriksson et al. 1998). In this review, we focus
on neurogenesis in the rodent brain.
In normal conditions, neurogenesis occurs in two
brain regions of adult rodents, the subventricular zone
*Author to whom all correspondence should be addressed.
Email: iimayosh@virus.kyoto-u.ac.jp
Received 25 November 2008; revised 17 December 2008;
accepted 02 January 2009.
© 2009 The Authors
Journal compilation © 2009 Japanese Society of
Developmental Biologists
(SGZ) of the hippocampal dentate gyrus. A huge number
of neurons born in the SVZ migrate through the rostral
migratory stream, and become local interneurons
(granule cells and periglomerular cells) in the olfactory
bulb, while neurons born in the SGZ migrate into the
granule cell layer and become granule cells of the
dentate gyrus. It has been shown that newly formed
neurons are incorporated into the functional networks
of both the olfactory bulb and the dentate gyrus,
suggesting a significant impact of adult neurogenesis
on brain functions (van Praag et al. 2002; Carleton et al.
2003; Kee et al. 2007).
Genetic labeling of neural stem cells
To study adult neurogenesis in vivo, it is necessary to
identify newly generated neurons among billions of
pre-existing neurons in the adult brain. Three major
approaches have been frequently adapted to label
newborn neurons (Ming & Song 2005; Gould 2007). One
is based on the incorporation of exogenous nucleotide
analogs, such as [H3]-thymidine and BrdU, into dividing
cells. During DNA replication, these nucleotide analogs
are incorporated into newly synthesized DNA of neural
stem cells (NSCs)/neural progenitor cells and inherited
to their progeny, such as new neurons. Another one
is genetic marking with retroviruses, which are only
infectious to dividing cells. The retroviral genome is
integrated into the host genome during the M-phase
of the cell cycle. Retroviruses carrying reporter genes,
380 I. Imayoshi et al.
such as green fluorescent protein (GFP) and LacZ, can
be used for specific labeling of dividing NSCs/progenitor
cells. However, these methods have only limited efficiency,
because [H3]-thymidine and BrdU can be administered
only for restricted periods, and because administration
of retroviruses into the brain using injection needles can
infect only small populations. A third one is a genetic
method using transgenic mice, in which reporter pro-
teins are expressed under the control of promoters of
immature neuron-specific genes such as doublecortin
(DCX) and proopiomelanocortin (POMC) (Overstreet
et al. 2004). However, these promoters are active
only transiently during neuronal differentiation, and it is
impossible to label newborn neurons permanently.
Therefore, we need more efficient methods that allow
selective and permanent marking of newborn neurons
in a temporally controlled manner.
Recently, a new method, the Cre/loxP system, has
been extensively used for fate-mapping analysis of
progenitor cells in various tissues (Nagy 2000; Kessaris
et al. 2005) (Fig. 1A). The bacteriophage P1 Cre recom-
binase efficiently excises DNA, which is flanked by two
directly repeated loxP recognition sites, in mammalian
cells. By crossing transgenic mice expressing Cre in
a cell type-specific manner with reporter mice, we can
trace the lineage of the progeny of Cre expressing cells
with reporter gene expression such as GFP and LacZ.
In reporter mice, a reporter gene is under the control of
an ubiquitous promoter such as CAG (CMV enhancer/
chicken β-actin promoter) and Rosa26 promoter, but the
expression is interrupted by a loxP-flanked transcrip-
tional STOP cassette. In these mice, recombination by
Cre results in the removal of the STOP cassette and
permanent expression of reporter protein. Temporal control
© 2009 The Authors
Journal compilation © 2009 Japanese Society of Developmental Biologists
Fig. 1. Genetic engineering of
neural stem cells (NSCs) by the
inducible Cre/loxP system. (A) Fate
mapping strategy to mark Cre-
expressing cells. Cre-expressing
transgenic animals are crossed
with the Rosa26-stop-reporter lines.
In Cre-expressing cells, the stop
cassette flanked by loxP sites is
excised, resulting in permanent
reporter expression. This genomic
recombination is transmitted to
their progeny. (B) Structure of the
Nes-CreERT2 transgene. Nestin
promoter and enhancer drive the
neural progenitor-specific expression
of CreERT2. Cre recombinase activity
is induced in neural progenitor cells
by the administration of tamoxifen.
of Cre-mediated recombination can be achieved by using
the ligand-dependent chimeric recombinase CreERT2.
CreERT2 is constructed by fusing Cre to the mutated
ligand-binding domain (LBD) of estrogen receptor
(Metzger et al. 1995; Feil et al. 1997). It has been shown
that in transgenic mice, CreERT2 is activated by admin-
istration of tamoxifen, a synthetic estrogen antagonist,
but not by natural ligands of LBD such as 17β-estradiol
(Indra et al. 2000; Li et al. 2000; Leone et al. 2003).
In order to mark NSCs and their progeny in the
adult brain, we generated Nes-CreERT2 transgenic mice
(Imayoshi et al. 2006), in which CreERT2 is expressed
under the control of the neural progenitor-specific nestin
promoter and enhancer (Zimmerman et al. 1994) (Fig. 1B).
In Nes-CreERT2 transgenic embryos, CreERT2 was spe-
cifically expressed in the ventricular zone of the developing
central nervous system and, in mice expressing CreERT2
at an appropriate level, Cre recombinase activity was
efficiently induced in neural stem cells by the adminis-
tration of tamoxifen (Imayoshi et al. 2006).
Nestin is expressed in the SVZ of the adult brain
(Doetsh et al. 1997), and in Nes-CreERT2 transgenic mice,
NSCs and transit-amplifying cells expressed CreERT2. In
the presence of tamoxifen, Cre-mediated recombination
occurred efficiently in NSCs, and the majority of newborn
neurons generated from such recombined NSCs were
labeled with reporter gene expression after tamoxifen
treatment (Imayoshi et al. 2008).
Replacement of granule cells in the olfactory
bulb
The SVZ is a layer extending along the lateral wall of
the lateral ventricle and contains many dividing cells
 Neurogenesis in adult brain
Fig. 2. Replacement of granule cells in the olfactory bulb by
newborn neurons. (A) Neurogenic niche of the subventricular zone
(SVZ). The SVZ is located near the lateral ventricle (LV) wall. In this
region, neural stem cells (NSCs) exist and new neurons are
continuously generated. Adult NSCs are glial fibrillary acidic protein
(GFAP)-positive cells with the structural and molecular characteristics
of astrocytes. NSCs divide to generate transit-amplifying cells
(Mash1-positive), which in turn differentiate into neuroblasts
(doublecortin [DCX]-positive) that migrate into the olfactory bulb.
(B) Newborn neurons reach the olfactory bulb (OB) through the
rostral migratory stream (RMS). (C) Most granule cells are replaced
by new neurons especially in deep regions of the granule cell layer
within one year. (D) Supply of new neurons is essential for
maintenance of the olfactory bulb. When olfactory neurogenesis is
blocked, extensive depletion of granule cells occurs in the olfactory
bulb.
(Fig. 2A). It was previously shown that a subset of glial
fibrillary acidic protein (GFAP)+ cells (Type B cells) func-
tion as neural stem cells in the adult SVZ (Doetsh et al.
1999). Type B cells divide slowly and give rise to rapidly
proliferating ‘transit-amplifying cells’ (Type C cells), which
then generate migrating neuroblasts (Type A cells)
after several cell divisions. The identification of the
SVZ cell types is mainly based on morphological anal-
ysis by electron microscopy and on the marker expres-
sion by immunohistochemistry (Doetsh et al. 1997,
Journal compilation © 2009 Japanese Society of Developmental Biologists
381
1999). Type B cells have the ultrastructural features of
astrocytes and express GFAP, a canonical astrocyte
marker protein. Type C and Type A cells can be identified
as GFAP-Mash1+Dlx2+ and GFAP-Dlx2+DCX+PSA-NCAM+
populations, respectively (Doetsh et al. 1997; Parras
et al. 2004). It has been reported that the ependymal
cells that surround the lateral ventricle (Type E cells)
may also be NSCs (Johansson et al. 1999), but several
studies have shown that ependymal cells are quiescent
and do not have the potentials of NSCs (Doetsh et al.
1999; Capela & Temple 2002).
Neuroblasts originating from the SVZ of adult rodents
migrate into the olfactory bulb, where they differentiate
into local interneurons (Lledo et al. 2006) (Fig. 2B).
Neuroblasts migrate through a path known as the
rostral migratory stream. This long distance migration
is not supported by radial glia or axon fibers as in the
embryonic brain, but through tubular structures formed by
specialized astrocytes (Lois et al. 1996). After neuroblasts
reach the core region of the olfactory bulb, they turn to
migrate radially to granule cell layers and periglomerular
layers, where they differentiate into mature interneurons.
Adhesion molecules (such as Poly-Sialated Neural Cell
Adhesion Molecule [PSA-NCAM] and tenascin-R) are
required for the proper organization of the rostral migratory
stream. Various chemoattractant and repellent signals
have been shown to regulate the direction of neuroblast
migration and the final destination of new neurons
(Lledo et al. 2006). Interestingly, the flow of cerebrospinal
fluid generated by beating of ependymal cilia appears
to generate gradients of Slit proteins for directing the
migration of neuroblasts (Sawamoto et al. 2006).
A huge number of neurons born in the SVZ continue
to migrate into the olfactory bulb. It has been suggested
that in the olfactory bulb old neurons are replaced by
new neurons, because the volume of the olfactory bulb
does not significantly change over the lifespan (Roselli-
Austin & Altman 1979; Kaplan et al. 1985; Petreanu &
Alvarez-Buylla 2002) and a relatively high-rate of apop-
tosis is observed in the granule cell layers (Biebl et al.
2000). However, it has not been clear to what extent
old granule cells are replaced by new neurons, and
whether the majority is replaced or only subsets of
granule cells are repeatedly replaced in the olfactory
bulb (Lledo et al. 2006).
Granule cells are axonless inhibitory interneurons
that have both a basal dendrite and an apical dendrite.
The apical dendrites make bidirectional dendrodendritic
synapses on their spines with the secondary dendrites
of mitral cells. These bidirectional synapses receive
glutamatergic input from the lateral dendrites of the
mitral or tufted cells (projection neurons of the olfactory
bulb) and release GABA back onto these projection
neurons (Mori & Shepherd 1994). These dendrodendritic
© 2009 The Authors
382 I. Imayoshi et al.
synapses are thought to be responsible for lateral
inhibition of the projection neurons in the olfactory bulb
(Yokoi et al. 1995).
Previous reports showed that olfactory discrimination
learning increases the survival of new neurons, and that
sensory experience during a critical period is important
for the survival of new neurons (Yamaguchi & Mori
2005; Alonso et al. 2006). These observations led to
the hypothesis that continual integration of new neurons
has important roles in the olfactory function, such as
olfactory discrimination and plasticity of olfactory circuits.
Several studies suggested that a supply of new
neurons is useful for plasticity and olfactory discrimina-
tion (Gheusi et al. 2000; Lledo et al. 2006). However,
the definite roles of olfactory interneuron replacement
have not been unveiled.
We crossed Nes-CreERT2 mice with Rosa26-stop-LacZ
and Rosa26-stop-CFP reporter mice and induced Cre
recombinase activity in adults with tamoxifen and
thereby efficiently labeled neural stem cells and their
progeny (Imayoshi et al. 2008). After tamoxifen treatment,
neuronal differentiation from recombined NSCs, and
migration and maturation of labeled new neurons into
the olfactory bulb were observed, as reported previously
(Petreanu & Alvarez-Buylla 2002). Long-term analysis
revealed that almost the entire granule cell population
was replaced by new neurons over a 12-month period
in the deep region of the olfactory bulb (Fig. 2C). In
contrast, in the superficial region, only half of the
neurons were replaced. It was reported that granule cells
born during an early postnatal stage predominantly
settle in the superficial region of the granule cell layer
and survive longer (Lemasson et al. 2005). Therefore,
deep and superficial granule cells have different turnover
rates, and the replacement mode is different between
superficial and deep regions.
Very little is known about synaptic outputs of new
granule cells. It remains to be determined whether a
new granule cell makes the synaptic contacts to the
same targets as the eliminated old granule cell, or to
the completely different targets. Further engineering
and application of in vivo multiphoton confocal imaging
might help to solve this issue (Mizrahi 2007).
We next asked whether death of old neurons is
induced by a supply of new neurons or old neurons
die even in the absence of new neurons. To address
this question, the majority of new neurons were ablated
by crossing Nes-CreERT2 mice with nuron-specific eno-
lase (NSE)-stop-DTA mice. In this double transgenic
mice, tamoxifen treatment in adults induces the DTA
(diphtheria toxin fragment A) expression from neuron-
specific enolase locus, and results in specific ablation
of newborn neurons (Imayoshi et al. 2008). Blockade of
the supply of new neurons led to gradual decrease of
© 2009 The Authors
Journal compilation © 2009 Japanese Society of Developmental Biologists
the granule cell number in the olfactory bulb (Fig. 2D).
Thus, granule cells have an intrinsic short lifetime, and
cell death occurs irrespective of a supply of new neu-
rons, indicating that continuous neurogenesis is essen-
tial for maintenance of the granule cell number in the
olfactory bulb. Recent study demonstrated that neuro-
nal elimination is an active process, rather than a simple
consequence of non-use, providing the vacancy for
coming new neurons (Mouret et al. 2008).
To assess whether the supply of new neurons is
required for discrimination and memory of odors, we
carried out olfactory discrimination tests using tamoxifen
treated Nes-CreERT2; NSE-stop-DTA mice (Imayoshi
et al. 2008). Surprisingly, blocking olfactory neurogenesis
had no effect on simple olfactory discrimination and
memory tests, although more difficult tasks about
odor-associated memory in different contexts could
depend on newborn neurons.
Periglomerular cells in the glomerular layer as well as
granule cells in the olfactory bulb are replaced by newborn
neurons (Kohwi et al. 2007). Periglomerular cells were
subdivided into at least three subtypes based on
immunoreactivity to tyrosine hydroxylase, calbindin and
calretinin (Kosaka et al. 1998), which show different
turnover rates (Kohwi et al. 2007; Ninkovic et al. 2007).
In mice, all three periglomerular cell subtypes seem
to be GABA-expressing inhibitory neurons, but the
physiological characteristics and functional roles of
each neuronal subtype have not been well determined.
Future studies using more sophisticated ablation
methods, such as interneuron subtype-specific
ablation, may unveil the functional significance of
replacement of each interneuron subtype in the adult
olfactory circuits.
Addition of granule cells in the dentate gyrus
In the SGZ of the adult brain, a subset of GFAP-positive
astrocytes has been shown to be progenitors for new
granule neurons (Seri et al. 2001, 2004) (Fig. 3A).
These GFAP-positive cells (Type I cells) have radial
processes extending through the entire granule cell
layer and short tangential processes in the molecular
layer. GFAP-negative progenitors, which have only
short processes (Type II cells), were also observed in
the SGZ (Suh et al. 2007). Definitive lineage relationship
between these two progenitors remains to be deter-
mined, but it seems that Type I cells give rise to Type II
cells (Fukuda et al. 2003). Nestin and Sox2 are expressed
by both progenitors (Suh et al. 2007). New neurons in
the SGZ migrate only a short distance into the inner
granule cell layer, and extend axons projecting through
the hilus region into the CA3 region (Zhao et al. 2006).
Recent studies showed that Reelin signaling and DISC1
 Neurogenesis in adult brain
Fig. 3. Addition of granule cells in
the hippocampal dentate gyrus. (A)
Adult neurogenesis in the dentate
gyrus (DG) of the hippocampus.
Type I and Type II progenitor cells
in the subgranular zone (SGZ) are
identified. These progenitor cells
give rise to transit-amplifying cells.
Then, transit-amplifying cells
differentiate into dentate granule
cells through several maturation
steps. (B) Adult neurogenesis
leads to addition of neurons to the
existing system in the dentate
gyrus. Newborn neurons are
mainly incorporated into the inner side of the granule cell layer. (C) When hippocampal neurogenesis is blocked, the existing neuronal circuit
is not significantly affected but the increase in the number of neurons is inhibited in the dentate gyrus.
regulate migration and positioning of newborn granule
cells (Duan et al. 2007; Gong et al. 2007). Granule cells
receive excitatory synaptic input from ascending fiber
tract originating from the entorhinal cortex.
Previous studies showed that few pre-existing neurons
die in the dentate gyrus, and that the total number of
granule cells of the dentate gyrus increases during
adulthood. Therefore, it has been suggested that
neurogenesis contributes to the increase in neuronal
number in the dentate gyrus (Bayer et al. 1982; Boss et al.
1985; Crespo et al. 1986; Dayer et al. 2003; Kempermann
et al. 2003). Tamoxifen treatment efficiently induced the
recombination in the SGZ progenitor cells of adult
Nes-CreERT2; Rosa26-stop-CFP mice, and labeled granule
cells gradually increased in number and expanded into
the granular cell layer, mostly in its inner side (Imayoshi
et al. 2008) (Fig. 3B). Labeled new neurons increased
to about 10% of the total granule cell number within
6 months after tamoxifen treatment, but this increase
stopped thereafter. This saturation is probably due to
decrease of neurogenesis in the SGZ of aged mice
(Kuhn et al. 1996). We also examined the effect of abla-
tion of neurogenesis in the dentate gyrus. Blockade of
neurogenesis inhibits the increase in the total number
of granule cells (Fig. 3C). This contrasts with control
mice, in which the number of granule cells increases
over the same period of time. Thus, neurogenesis in
the dentate gyrus led to addition of neurons to the
existing system, and without neurogenesis, the structure
was maintained but the increase in the granule cell
number was inhibited (Fig. 3). The pre-existing neuronal
circuit does not seem to be significantly affected by
blockade of neurogenesis. This is a sharp contrast to the
olfactory bulb (Fig. 2), where inhibition of neurogenesis
Journal compilation © 2009 Japanese Society of Developmental Biologists
383
resulted in substantial reduction of the granule cell
number. These results suggest different roles and
integration modes for adult neurogenesis: maintenance
and reorganization of the whole interneuron system in the
olfactory bulb; modulation and refinement of existing
neuronal circuits in the dentate gyrus.
To assess whether neurogenesis in the dentate gyrus is
required for hippocampus-dependent memory, tamoxifen-
treated Nes-CreERT2; NSE-stop-DTA mice were trained for
spatial and fear learning tasks. Blockade of neurogenesis
was found to impair retention of spatial memory and
formation of contextual memory. These results agreed
well with the previous reports (Snyder et al. 2005; Saxe
et al. 2006). However, the precise mechanism of how
new neurons contribute to the hippocampus-dependent
memory is still unclear. As in the case of the olfactory
bulb, very little is known about synaptic inputs and outputs
of new dentate granule cells. A possible hypothesis is
that newborn granule cells are preferentially integrated into
special hippocampal circuits. Place cells in hippocampal
CA1 area probably receive spatial information from the
entorhinal cortex and CA3 area. Interestingly, direct
input from the entorhinal cortex is sufficient for estab-
lishing fundamental properties of place cells in the CA1
area, and the input from the dentate gyrus-CA3 system
is required for the efficient associative recall of spatial
memory (Brun et al. 2002; Nakazawa et al. 2002).
Blockade of neurogenesis impaired recall, but not
acquisition, of spatial memory after a certain time
passed (Imayoshi et al. 2008). Detailed analysis of the
effect of continuous neuronal addition on the plasticity
of hippocampal circuits by electrophysiological studies will
define how neurogenesis contributes to the hippocampus-
dependent memory.
© 2009 The Authors
384 I. Imayoshi et al.
Concluding remarks
Since the discovery of adult neurogenesis, many extensive
studies have been carried out on various aspects of this
phenomenon, including proliferation, fate-specification
of adult neural stem cells, and migration, maturation and
synaptic integration of new neurons. Postdevelopmental
neurogenesis is found to be an evolutionarily conserved
phenomenon, and functional importance on brain
activities has just begun to be unveiled. Further under-
standing of adult neurogenesis will help the application
of cell replacement therapy to the damaged brain after
injury or neurological diseases. Especially, it is very
important to understand what is happening on existing
neural circuits and brain functions when exogenous
neurons are added. Full understanding of the regulatory
mechanism and functional roles of endogenous adult
neurogenesis will help us develop the sophisticated
strategy of regenerative medicine for neurological diseases
in humans.
Acknowledgments
This work was supported by Grants-in-aid from the
Ministry of Education, Culture, Sports, Science and
Technology of Japan. I. Imayoshi was supported by
Research Fellowships of the Japan Society for the
Promotion of Science for Young Scientists. We thank
Masahiro Yamaguchi, Kensaku Mori, Keizo Takao,
Tsuyoshi Miyakawa for valuable discussion.
Conflict of Interest
No conflict of interest has been declared by I. Imayoshi,
M. Sakamoto, T. Ohtsuka or R. Kageyama.
References
Alonso, M., Viollet, C., Gabellec, M. M., Meas-Yedid, V., Olivo-Marin,
J. C. & Lledo, P. M. 2006. Olfactory discrimination learning
increases the survival of adult-born neurons in the olfactory
bulb. J. Neurosci. 26, 10508–10513.
Altman, J. 1966. Autoradiographic and histological studies of
postnatal neurogenesis. II. A longitudinal investigation of the
kinetics, migration and transformation of cells incorporating
tritiated thymidine in infant rats, with special reference to
postnatal neurogenesis in some brain regions. J. Comp.
Neurol. 1966, 431–474.
Altman, J. 1969. Autoradiographic and histological studies of
postnatal neurogenesis. IV. Cell proliferation and migration in the
anterior forebrain, with special reference to persisting neuro-
genesis in the olfactory bulb. J. Comp. Neurol. 137, 433–457.
Altman, J. & Das, G. D. 1965. Autoradiographic and histological
evidence of postnatal hippocampal neurogenesis in rats. J.
Comp. Neurol. 124, 319–335.
Alvarez-Buylla, A., Garcia-Verdugo, J. M. & Tramontin, A. D. 2001.
A unified hypothesis on the lineage of neural stem cells. Nat.
Rev. Neurosci. 2, 287–293.
© 2009 The Authors
Journal compilation © 2009 Japanese Society of Developmental Biologists
Bayer, S. A., Yackel, J. W. & Puri, P. S. 1982. Neurons in the rat
dentate gyrus granular layer substantially increase during
juvenile and adult life. Science 216, 890–892.
Biebl, M., Cooper, C. M., Winkler, J. & Kuhn, H. G. 2000. Analysis
of neurogenesis and programmed cell death reveals a self-
renewing capacity in the adult rat brain. Neurosci. Lett. 291,
17–20.
Boss, B. D., Peterson, G. M. & Cowan, M. 1985. On the number
of neurons in the dentate gyrus of the rat. Brain Res. 338,
144–150.
Brun, V. H., Otnass, M. K., Molden, S., Steffenach, H. A., Witter,
M. P., Moser, M. B. & Moser, E. I. 2002. Place cells and place
recognition maintained by direct entorhinal-hippocampal
circuitry. Science 296, 2243–2246.
Capela, A. & Temple, S. 2002. LeX/ssea-1 is expressed by adult
mouse CNS stem cells, identifying them as nonependymal.
Neuron 35, 865–875.
Carleton, A., Petreanu, L. T., Lansford, R., Alvarez-Buylla, A. &
Lledo, P. M. 2003. Becoming a new neuron in the adult
olfactory bulb. Nat. Neurosci. 6, 507–518.
Crespo, D., Stanfield, B. B. & Cowan, W. M. 1986. Evidence that
late-generated granule cells do not simply replace earlier
formed neurons in the rat dentate gyrus. Exp. Brain Res. 62,
541–548.
Dayer, A. G., Ford, A. A., Cleaver, K. M., Yassaee, M. & Cameron,
H. 2003. Short-term and long-term survival of new neurons in
the rat dentate gyrus. J. Comp. Neurol. 460, 563–572.
Doetsch, F. 2003. The glial identity of neural stem cells. Nat.
Neurosci. 6, 1127–1134.
Doetsh, F., Caille, I., Lim, D. A., Garcia-Verdugo, J. M. & Alvarez-
Buylla, A. 1999. Subventricular zone astrocytes are neural
stem cells in the adult mammalian brain. Cell 97, 703–716.
Doetsh, F., Garcia-Verdugo, J. M. & Alvarez-Buylla, A. 1997.
Cellular composition and three-dimensional organization of
the subventricular germinal zone in the adult mammalian
brain. J. Neurosci. 17, 5046–5061.
Gong, C., Wang, T., Huang, H. S. & Parent, J. M. 2007. Reelin
regulates Neuronal Progenitor Migration in intact and epileptic
hippocampus. J. Neurosci. 27, 1803–1811.
Duan, X., Chang, J. H., Ge, S., Faulkner, R. L., Kim, J. Y.,
Kitabatake, Y., Liu, X. B., Yang, C. H., Jordan, J. D., Ma, D.
K., Liu, C. Y., Ganesan, S., Cheng, H. J., Ming, G. L., Lu, B.
& Song, H. 2007. Disrupted-In-Schizophrenia 1 regulates
integration of newly generated neurons in the adult brain. Cell
130, 1146–1158.
Eriksson, P. S., Perfililieva, E., Bjork-Eriksson, T., Alborn, A. M.,
Nordborg, C., Peterson, D. A. & Gage, F. H. 1998. Neurogenesis
in the adult human hippocampus. Nat. Med. 4, 1313–1317.
Feil, R., Wagner, J., Metzger, D. & Chambon, P. 1997. Regulation
of Cre recombinase activity by mutated estrogen receptor
ligand-binding domains. Biochem. Biophys. Res. Commun.
237, 752–757.
Fukuda, S., Kato, F., Tozuka, Y., Yamaguchi, M., Miyamoto, Y.
& Hisatsune, T. 2003. Two distinct subpopulations of nestin-
positive cells in adult mouse dentate gyrus. J. Neurosci. 23,
9357–9366.
Gage, F. H. 2000. Mammalian neural stem cells. Science 287,
1433–1438.
Gheusi, G., Cremer, H., McLean, H., Chazal, G., Vincent, J. D. &
Lledo, P. M. 2000. Importance of newly generated neurons in
the adult olfactory bulb for odor discrimination. Proc. Natl
Acad. Sci. USA 97, 1823–1828.
Gong, C., Wang, T., Huang, H. S. & Parent, J. M. 2007. Reelin
regulates neuronal progenitor migration in intact and epileptic
hippocampus. J. Neurosci. 27, 1803–1811.
 Neurogenesis in adult brain
Gould, E. 2007. How widespread is adult neurogenesis in mammals?
Nat. Rev. Neurosci. 8, 481–488.
Imayoshi, I., Ohtsuka, T., Metzger, D., Chambon, P. & Kageyama,
R. 2006. Temporal regulation of Cre recombinase activity in
neural stem cells. Genesis 44, 233–238.
Imayoshi, I., Sakamoto, M., Ohtsuka, T., Takao, K., Miyakawa, T.,
Yamaguchi, M., Mori, K., Ikeda, T., Itohara, S. & Kageyama,
R. 2008. Roles of continuous neurogenesis in the structural
and functional integrity of the adult forebrain. Nat. Neurosci.
11, 1153–1161.
Indra, A. K., Li, M., Brocard, J., Warot, X., Bornert, J. M., Gérard,
C., Messaddeq, N., Chambon, P. & Metzger, D. 2000.
Targeted somatic mutagenesis in mouse epidermis. Horm.
Res. 54, 296–300.
Johansson, C. B., Momma, S., Clarke, D. L., Risling, M., Lendahl,
U. & Frisen, J. 1999. Identification of a neural stem cell in the
adult mammalian central nervous system. Cell 96, 25–34.
Kaplan, M. S. & Hinds, J. W. 1977. Neurogenesis in the adult rat:
electron microscopic analysis of light radioautographs.
Science 197, 1092–1094.
Kaplan, M. S., McNelly, N. A. & Hinds, J. W. 1985. Population
dynamics of adult-formed granule neurons of the rat olfactory
bulb. J. Comp. Neurol. 239, 117–125.
Kee, N., Teixeira, C. M., Wang, A. H. & Frankland, P. W. 2007.
Preferential incorporation of adult-generated granule cells into
spatial memory networks in the dentate gyrus. Nat. Neurosci.
10, 355–362.
Kempermann, G., Gast, D., Kronenberg, G., Yamaguchi, M. & Gage,
F. H. 2003. Early determination and long-term persistence of
adult-generated new neurons in the hippocampus of mice.
Development 130, 391–399.
Kessaris, N., Fogarty, M., Iannnarelli, P., Grist, M., Wegner, M. &
Richardson, W. D. 2005. Competing waves of oligodendro-
cytes in the forebrain and postnatal elimination of an embryonic
lineage. Nat. Neurosci. 9, 173–179.
Kohwi, M., Petryniak, M. A., Long, J. E., Ekker, M., Obata, K.,
Yanagawa, Y., Rubenstein, J. L. & Alvarez-Buylla, A. 2007. A
subpopulation of olfactory bulb GABAergic interneurons is
derived from Emx1- and Dlx5/6-expressing progenitors. J.
Neurosci. 27, 6878–6891.
Kosaka, K., Toida, K., Aika, Y. & Kosaka, T. 1998. How simple is
the organization of the olfactory glomerulus? The heterogeneity
of so-called periglomerular cells. Neurosci. Res. 30, 101–110.
Kuhn, H. G., Dickinson-Anson, H. & Gage, F. H. 1996. Neurogenesis
in the dentate gyrus of the adult rat: age-related decrease
of neuronal progenitor proliferation. J. Neurosci. 16, 2027–
2033.
Lemasson, M., Saghatelyan, A., Olivo-Marin, J. C. & Lledo, P. M.
2005. Neonatal and adult neurogenesis provide two distinct
populations of newborn neurons to the mouse olfactory bulb.
J. Neurosci. 25, 6816–6825.
Leone, D. P., Genoud, S., Atanasoski, S., Grausenburger, R.,
Berger, P., Metzger, D., Macklin, W. B., Chambon, P. & Suter,
U. 2003. Tamoxifen-inducible glia-specific Cre mice for
somatic mutagenesis in oligodendrocytes and Schwann cells.
Mol. Cell. Neurosci. 22, 430–440.
Li, M., Indra, A. K., Warot, X., Brocard, J., Messaddeq, N., Kato,
S., Metzger, D. & Chambon, P. 2000. Skin abnormalities
generated by temporally controlled RXRalpha mutations in
mouse epidermis. Nature 407, 633–636.
Lledo, P. M., Alonso, M. & Grubb, M. S. 2006. Adult neurogenesis
and functional plasticity in neuronal circuits. Nat. Rev. Neurosci.
7, 179–193.
Lois, C., Garcia-Verdugo, J. M. & Alvarez-Buylla, A. 1996. Chain
migration of neuronal precursors. Science 271, 978–981.
Journal compilation © 2009 Japanese Society of Developmental Biologists
385
McKay, R. 1997. Stem cell in the central nervous system. Science
276, 66–71.
Metzger, D., Clifford, J., Chiba, H. & Chambon, P. 1995. Conditional
site-specific recombination in mammalian cells using a
ligand-dependent chimeric Cre recombinase. Proc. Natl Acad.
Sci. USA 92, 6991–6995.
Ming, G. & Song, H. 2005. Adult neurogenesis in the mammalian
central nervous system. Annu. Rev. Neurosci. 28, 223–250.
Mizrahi, A. 2007. Dendritic development and plasticity of adult-born
neurons in the mouse olfactory bulb. Nat. Neurosci. 10, 444–
452.
Mori, K. & Shepherd, G. M. 1994. Emerging principles of molecular
signal processing by mitral/tufted cells in the olfactory bulb.
Semin. Cell Biol. 5, 65–74.
Mouret, A., Gheusi, G., Gabellec, M. M., Chaumont, F. D., Olivo-Marin,
J. C. & Lledo, P. M. 2008. Learning and survival of newly
generated neurons: when time matters. J. Neurosci. 28,
11511–11516.
Nagy, A. 2000. Cre recombinase: the universal reagent for
genome tailoring. Genesis 26, 99–109.
Nakazawa, K., Quirk, M. C., Chitwood, R. A., Watanabe, M., Yeckel,
M. F., Sun, L. D., Kato, A., Carr, C. A., Johnston, D., Wilson,
M. A. & Tonegawa, S. 2002. Requirement for hippocampal
CA3 NMDA receptors in associative memory recall. Science
297, 211–218.
Ninkovic, J., Mori, T. & Götz, M. 2007. Distinct modes of neuron
addition in adult mouse neurogenesis. J. Neurosci. 27,
10906–10911.
Overstreet, L. S., Hentges, S. T., Bumaschny, V. F., de Souza, F. S.
J., Smart, J. L., Santangelo, A. M., Low, M. J., Westbrook, G.
L. & Rubinstein, M. 2004. A transgenic marker for newly
born granule cells in dentate gyrus. J. Neurosci. 23, 3251–
3259.
Parras, C. M., Galli, R., Britz, O., Soares, S., Galichet, C., Battiste,
J., Johnson, J. E., Nakafuku, M., Vescovi, A. & Guillemot, F.
2004. Mash1 specifies neurons and oligodendrocytes in the
postnatal brain. EMBO J. 23, 4495–4505.
Petreanu, L. & Alvarez-Buylla, A. 2002. Maturation and death of
adult-born olfactory bulb granule neurons: role of olfaction. J.
Neurosci. 22, 6106–6133.
van Praag, H., Schinder, A. F., Christie, B. R., Toni, N., Palmer, T.
D. & Gage, F. H. 2002. Functional neurogenesis in the adult
hippocampus. Nature 415, 1030–1034.
Roselli-Austin, L. & Altman, J. 1979. The postnatal development
of the main olfactory bulb of the rat. J. Dev. Physiol. 1, 295–
313.
Sawamoto, K., Wichterle, H., Gonzalez-Perez, O., Cholfin, J. A.,
Yamada, M., Spassky, N., Murcia, N. S., Garcia-Verdugo, J.
M., Marin, O., Rubenstein, J. L. R., Tessier-Lavigne, M.,
Okano, H. & Alvarez-Buylla, A. 2006. New neurons follow the
flow of cerebrospinal fluid in the adult brain. Science 311,
629–632.
Saxe, M. D., Battaglia, F., Wang, J. W., Malleret, G., David, D. J.,
Monckton, J. E., Garcia, A. D., Sofroniew, M. V., Kandel, E.
R., Santarelli, L., Hen, R. & Drew, M. R. 2006. Ablation of
hippocampal neurogenesis impairs contextual fear conditioning
and synaptic plasticity in the dentate gyrus. Proc. Natl Acad.
Sci. USA 103, 17501–17506.
Seri, B., Garcia-Verdugo, J. M., McEwen, B. S. & Alvarez-Buylla,
A. 2001. Astrocytes give rise to new neurons in the adult
mammalian hippocampus. J. Neurosci. 21, 7153–7160.
Seri, B., Garcia-Verdugo, J. M., Collado-Morente, L., McEwen, B. S.
& Alvarez-Buylla, A. 2004. Cell types, lineage, and architecture
of the germinal zone in the adult dentate gyrus. J. Comp.
Neurol. 478, 359–378.
© 2009 The Authors
386 I. Imayoshi et al.

Snyder, J. S., Hong, N. S., McDonald, R. J. & Wojtowicz, J. M.
2005. A role for adult neurogenesis in spatial long-term
memory. Neuroscience 130, 843–852.
Suh, H., Consiglio, A., Ray, J., Sawai, T., D-Amour, K. A. & Gage,
F. H. 2007. In vivo fate analysis reveals the multipotent and
self-renewal capacities of Sox2+ neural stem cells in the adult
hippocampus. Cell Stem Cell 1, 515–528.
Temple, S. 2001. The development of neural stem cells. Nature
414, 112–117.
Yamaguchi, M. & Mori, K. 2005. Critical period for sensory
experience-dependent survival of newly generated granule
cells in the adult mouse olfactory bulb. Proc. Natl Acad. Sci.
USA 102, 9697–9702.
Yokoi, M., MorI, K. & NakanishI, S. 1995. Refinement of odor
molecule tuning by dendrodendritic synaptic inhibition in
the olfactory bulb. Proc. Natl Acad. Sci. USA 92, 3371–
3375.
Zhao, C., Teng, E. M., Summers, R. G. Jr, Ming, G. L. & Gage, F.
H. 2006. Distinct morphological stages of dentate granule
neuron maturation in the adult mouse hippocampus. J.
Neurosci. 26, 3–11.
Zimmerman, L., Lendahl, U., Cunningham, M., McKay, R., Parr,
B., Gavin, B., Mann, J., Vassileva, G. & McMahon, A. 1994.
Independent regulatory elements in the nestin gene direct
transgene expression to neural stem cells or muscle precursoprs.
Neuron 12, 11–24.

© 2009 The Authors

Journal compilation © 2009 Japanese Society of Developmental Biologists