Archives of Biochemistry and Biophysics 611 (2016) 93e99

Contents lists available at ScienceDirect

Archives of Biochemistry and Biophysics

journal homepage: www.elsevier.com/locate/yabbi

Insight into cognitive decline from Zn2þ dynamics through

extracellular signaling of glutamate and glucocorticoids
Atsushi Takeda*, Hanuna Tamano
Department of Neurophysiology, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan

a r t i c l e i n f o a b s t r a c t

Article history: Glutamatergic neuron activity and/or the modification of the activity with glucocorticoids are closely
Received 1 April 2016 linked to synaptic Zn2þ dynamics as well as synaptic Ca2þ dynamics. The dynamic crosstalk of synaptic
Received in revised form Zn2þ signaling to intracellular Ca2þ signaling via calcium channels is involved in synaptic plasticity such
23 June 2016
as long-term potentiation (LTP) and cognitive activity. The influx of extracellular Zn2þ into postsynaptic
Accepted 30 June 2016
neurons, which is closely linked to glutamate signaling in the synaptic cleft, is critical for cognitive ac-
Available online 5 July 2016
tivity. However, excess intracellular Zn2þ signaling induced by excess glutamatergic neuron activity is
involved in not only cognitive decline in neurological disorders but also stress-induced cognitive decline.
Keywords:
Zinc
On the other hand, it has been recognized that excess Ca2þ influx into postsynaptic neurons induces
Glutamate neuronal death, while the involvement of excess intracellular Ca2þ signaling in cognitive decline is poorly
Glucocorticoid understood. Understanding of synaptic Zn2þ dynamics, which are modified by glutamate and gluco-
Hippocampus corticoid signaling, may be meaningful to prevent Zn2þ-mediated cognitive decline. This paper sum-
Cognition marizes the current knowledge on Zn2þ dynamics under changing synaptic environment and its impact
Calcium on cognitive decline.
© 2016 Elsevier Inc. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
2. Brain zinc homeostasis through the brain barrier system and its impact on cognition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
3. Impact on cognition from extracellular and intracellular Zn2þ homeostasis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
4. Impact on cognition from neuronal death via excess synaptic Zn2þ signaling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
5. Impact on cognition from synaptic Zn2þ signaling under diverse synaptic environment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
6. Perspective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98

1. Introduction responsible for the transport of Zn2þ out of the cytoplasm, are
involved in the absorption and excretion of Zn2þ to maintain zinc
Plasma zinc is thought to be a major pool of zinc in the living homeostasis in the living body [2e4].
body to transfer zinc to the tissues and organs including the brain. Zinc concentration in the plasma is approximately 15 mM. Zinc
Zinc homeostasis in the living body is tightly controlled by both transport from the plasma to the brain extracellular fluid and the
intestinal absorption and intestinal and renal excretions [1]. Zrt-Irt- cerebrospinal fluid (CSF) is regulated by the brain barrier system,
like proteins (ZIP), which are responsible for the movement of Zn2þ i.e., the bloodebrain and bloodeCSF barriers. Zinc concentration in
into the cytoplasm, and the zinc transporter (ZnT) family, which are the CSF is approximately 0.15e0.38 mM [5e7] and extracellular zinc
concentration in the brain is estimated to be less than 1 mM [8]. Zinc
concentration in the brain increases along with the development
* Corresponding author. and reaches 13.3 ± 0.3 mg/g wet brain in the human adult brain
E-mail address: takedaa@u-shizuoka-ken.ac.jp (A. Takeda). (infant brain, 8.2 ± 0.8 mg/g wet brain) [9]. Brain zinc homeostasis is

http://dx.doi.org/10.1016/j.abb.2016.06.021
0003-9861/© 2016 Elsevier Inc. All rights reserved.
94 A. Takeda, H. Tamano / Archives of Biochemistry and Biophysics 611 (2016) 93e99
strictly controlled through the brain barrier system under healthy
condition [10,11], while synaptic Zn2þ movement is changed by
synaptic activity, especially glutamatergic synapse activity and is
involved in synaptic function [12]. Approximately 80% of the total
brain zinc functions as zinc metalloproteins and the remaining part
is histochemically reactive as revealed by Timm’s sulfide-silver
staining method [13,14] and serves as a signal factor (in the form
of free Zn2þ) in the cytosolic compartment as well as the extra-
cellular compartment.
The hippocampus and amygdala are involved in cognitive and
emotional behavior and strongly stained by Timm’s method
[15e17]. To process the information on memory, glutamatergic
neurons compose the principal neural circuits in the hippocampus
and amygdala. The role of the neural circuits has been extensively
studied in the hippocampus. Memory are linked to strengthening of
synaptic connections between neurons, i.e., synaptic plasticity such
as long-term potentiation (LTP). Glutamatergic neuron activity is
closely linked to synaptic Zn2þ dynamics as well as synaptic Ca2þ
dynamics [15,16]. The dynamic crosstalk of synaptic Zn2þ signaling
to intracellular Ca2þ signaling via calcium channels is involved in
cognitive activity based on synaptic plasticity. Because a subclass of
glutamatergic (zincergic) neurons releases Zn2þ into the synaptic
cleft [13], synaptic (extracellular) Zn2þ movement is closely linked
to glutamate signaling in the synaptic cleft and modifies intracel-
lular Zn2þ signaling [18]. Even at non-zincergic synapses, intracel-
lular Zn2þ signaling, which mainly originates in the internal stores
containing Zn2þ, is critical for cognitive activity.
The hypothalamic-pituitary-adrenal (HPA) axis serves to main-
tain homeostasis in the living body and the HPA axis activity, i.e.,
glucocorticoid secretion, is enhanced after exposure to stress.
Glucocorticoids can potentiate glutamatergic neuron activity under
both pathological and physiological conditions [19,20] and modify
synaptic Zn2þ signaling [21]. Excess intracellular Zn2þ signaling
induced by excess glutamatergic neuron activity is involved in not
only cognitive decline in neurological disorders but also stress-
induced cognitive decline [22]. Thus, diverse Zn2þ dynamics
induced by glutamate and glucocorticoid signaling may modify
cognitive activity. To prevent Zn2þ-mediated cognitive decline, this
paper summarizes the current knowledge on Zn2þ dynamics under
changing synaptic environment and its impact on cognitive decline.
The present topics propose that the influx of extracellular Zn2þ into
postsynaptic neurons may be critical for cognitive decline in com-
parison with that of extracellular Ca2þ and probably involved in
age-related cognitive decline.
2. Brain zinc homeostasis through the brain barrier system
and its impact on cognition
Healthy brain maintains a very stable environment via the brain
barrier system, which is critical for brain function including
cognition. Zn2þ is slowly transported into the brain through the
bloodebrain and the bloodeCSF barriers [23,24]. The cells forming
the brain barrier system express ZIP and ZnT, and play key roles for
Zn2þ transport into the brain parenchyma and also brain zinc ho-
meostasis [25]. However, the mechanism on Zn2þ transport across
the brain barrier system is poorly understood and the mechanism
on brain zinc homeostasis is unclear [26,27]. On the other hand,
chronic zinc deficiency suppresses the increase in zinc concentra-
tion in the rat hippocampus, indicating that hippocampal zinc is
relatively susceptible to zinc deficiency in the brain [28]. Chronic
zinc deficiency also suppresses the increase in vesicular zinc. Brain
Zn2þ including hippocampal Zn2þ seems to be susceptible to
chronic zinc deficiency. The decrease in plasma zinc elicited by
dietary zinc deficiency may lead to the decrease in CSF zinc, which
is important for Zn2þ transport into the hippocampus [24]. Brain
Zn2þ homeostasis is critical for brain function such as cognitive
activity and seems to be modified along with brain aging [29e31].
Brain aging is characterized by neuronal loss, cognitive decline,
and susceptibility to neurological disorders [32]. The changes
occurring in the brain during aging may be often related to the
modification of Zn2þ homeostasis. Zinc transporter-3 (ZnT3) pro-
tein is responsible for loading zinc into presynaptic vesicles [33]
and consequently controls the availability of Zn2þ at zincergic
synapses, which is critical for hippocampus-dependent memory as
described later [34]. Because ZnT3 protein decreases along with
aging [35], it may be involved in age-related cognitive decline.
Adlard et al. [36,37] report that metal chaperones i.e., clioquinol
and PBT2, are transported into the brain through the brain barrier
system, can increase the availability of Zn2þ, and prevent normal
age-related cognitive decline. They demonstrate that metal chap-
erones are effective for preventing Zn2þ-mediated cognitive decline
that might be increased with aging. Metal chaperones transported
into the brain may be also effective as a therapeutic strategy for
neurodegenerative diseases associated with brain Zn2þ dysho-
meostasis [38,39] Brain Zn2þ dyshomeostasis may be involved in
cognitive decline under both pathological and physiological
conditions.
Zinc concentration in the brain is hardly affected after exposure
to zinc. The brain barrier system protects brain parenchyma cells by
blocking excess Zn2þ influx. In a bloodebrain barrier model, the
expressions of zinc transporters such as ZnT1 and ZnT2, and also
metallothioneins, zinc-binding proteins are changed to respond to
moderately excessive zinc environment [40]. The brain barrier
system protects brain parenchyma cells from neurotoxicity of
metals including zinc. The bloodebrain barrier breakdown that
occurs during normal aging is associated with inflammation and
disruption of tight junction complex assembly but not through
leukocyte trafficking [41]. Montagne et al. report that the bloode-
brain barrier breakdown is an early event in the aged human brain
that begins in the hippocampus and may contribute to cognitive
decline [42]. Furthermore, the brain capillary endothelial cells play
an active part in the process of neurological disorders, in which
cognitive decline is involved. The bloodebrain barrier is modified
and/or disrupted in the process of neurological disorders such as
neoplasia, stroke/ischemia, epilepsy, and dementia [43]. The in-
crease in the bloodebrain barrier permeability leads to the dys-
homeostasis of essential metals such as zinc and iron, which has
been implicated in many neurodegenerative disorders, followed by
cognitive decline [44].
3. Impact on cognition from extracellular and intracellular
Zn2þ homeostasis
The basal (static) concentration of Zn2þ in the brain extracellular
fluid is estimated to be 10 nM [45]. Fluorescence intensity of
extracellular ZnAF-2, an extracellular Zn2þ indicator [46], varies
among areas in the hippocampus [47,48], where the stratum luci-
dum and the hilus are strongly stained with ZnAF-2. At zincergic
synapses such as mossy fiber and Schaffer collateral synapses,
extracellular Zn2þ levels are increased by cognitive activity and LTP
induction. However, the optimal range (level) of Zn2þ in the
extracellular compartment remains to be clarified [49]. In vivo CA1
LTP is significantly attenuated under local perfusion of the
recording region with 100 nM ZnCl2 by using a recording electrode
attached to a microdialysis probe [50]. In the microdialysis exper-
iment, it is estimated that extracellular Zn2þ level in the recording
region is less than 100 nM, because the perfusate reaches the
equilibrium with extracellular fluid. When the basal level of
extracellular Zn2þ rises to several times by zincergic excitation in
the CA1 and the rise is maintained for a period of time, CA1 LTP can
 A. Takeda, H. Tamano / Archives of Biochemistry and Biophysics 611 (2016) 93e99
be affected. On the other hand, extracellular Zn2þ concentration
during tetanic stimulation to induce LTP is unclear at zincergic
synapses. It is possible that the concentration reaches micromolar
levels based on the estimated concentration of vesicular zinc
[51,52].
The basal Zn2þ concentrations are extremely low in the intra-
cellular (cytosol) compartment and estimated to be less than 1 nM
[53,54]. Intracellular Zn2þ homeostasis is closely linked to extra-
cellular Zn2þ dynamics even at non-zincergic synapses because the
basal concentration of intracellular Zn2þ is much lower than that of
extracellular Zn2þ. Fluorescence intensity of intracellular ZnAF-2
after addition of ZnAF-2DA, an intracellular Zn2þ indicator [46],
also varies among areas in the hippocampus [47,48], and stratum
lucidum and the hilus, which contain mossy fibers, are strongly
stained with ZnAF-2. It is likely that the basal levels of intracellular
Zn2þ are correlated with zinc concentration in the synaptic vesicle
[33]. There is the close correlation between vesicular Zn2þ level and
ZnT3 protein expression [55]. Zn2þ level other than Zn2þ released
from the synaptic vesicle is low and estimated to be less than 5% of
the total amount of Zn2þ in the hippocampus and cerebral cortex
[55]. At zincergic synapses, transsynaptic influx of Zn2þ released
from neuron terminals is involved in hippocampus-dependent
memory [50,56,57].
ZnT proteins such as ZnT1, ZnT3 and ZnT10, and ZIP such as
ZIP4 and ZIP6 are involved in the control of Zn2þ level in the
cytosolic compartment, especially under the basal circumstance
[58]. Some of these transporters transport cytosolic Zn2þ into
different subcellular organelles, e.g., mitochondria, lysosomes,
endosomes, and Golgi apparatus, probably to maintain the basal
Zn2þ level in the cytosolic compartment [59e61]. On the other
hand, Zn2þ release from the subcellular organelles is involved in
intracellular Zn2þ signaling [62]. Intracellular Ca2þ signaling via
glutamate receptor activation may play a key role for intracellular
Zn2þ signaling, although the mechanism is unknown. At non-
zincergic synapses, intracellular Zn2þ signaling via glutamate re-
ceptor activation is involved in hippocampus-dependent memory
[63].
ZnT1 is a major Zn2þ transporter on the plasma membrane
and involved in cytosolic Zn2þ homeostasis in neurons by
transporting Zn2þ from the somata to the extracellular space [64].
It has been reported that ZnT1 restricts excessive accumulation of
Zn2þ in the cytosolic compartment [65], resulting in protecting
neurons from Zn2þ toxicity in neurological disorders such as
transient forebrain ischemia [66]. Furthermore, ZnT1 concen-
trates at the postsynaptic density of hippocampal synapses
regardless of the presence of vesicular zinc, suggesting that ZnT1
is involved in hippocampus-dependent memory via maintaining
intracellular Zn2þ homeostasis [67]. ZIP1 and ZIP3 may provide
selective therapeutic targets to protect hippocampal neurons
from early zinc-induced neurodegeneration following injury [68].
Tissue plasminogen activator, a secreted serine protease is exci-
totoxic and increases lysosomal sequestration of the increased
Zn2þ in the cytosolic compartment through interaction with ZIP4,
which might also lead to protecting neurons from Zn2þ toxicity
[58]. The spatiotemporal control of intracellular Zn2þ via ZIP and
ZnT is involved in the steady environment of Zn2þ in the cytosolic
compartment, which is critical for hippocampus-dependent
memory [29]. The hippocampus, especially the dentate gyrus is
the most susceptible to aging in the brain [69]. If one of the
causes is the bloodebrain barrier breakdown in the hippocampus
[42], the control of ZIP and ZnT protein expression might be a key
for the strategy preventing age-related hippocampus-dependent
cognitive decline.
95
4. Impact on cognition from neuronal death via excess
synaptic Zn2þ signaling
Glutamatergic neuron activity plays a key role for cognition
under physiological and pathological conditions. Glutamate re-
ceptor activation by excess extracellular glutamate leads to a
number of deleterious consequences, including impairment of
calcium buffering, generation of free radicals, activation of the
mitochondrial permeability transition and secondary excitotoxicity
[70,71] and is known as glutamate excitotoxicity. It is a final com-
mon pathway for neuronal death and is observed in numerous
pathological processes such as stroke/ischemia, temporal lobe ep-
ilepsy, Alzheimer’s disease and amyotrophic lateral sclerosis
[72,73], which are often accompanied by cognitive impairment.
Neuronal death is induced by excess Zn2þ influx into postsynaptic
neurons, in addition to excess Ca2þ influx, at zincergic synapses and
is a cause for cognitive decline (Fig. 1) [74,75]. At non-zincergic
synapses, on the other hand, there is no report on Zn2þ-mediated
neuronal death. It is possible that Zn2þ influx into non-zincergic
postsynaptic neurons, in addition to Ca2þ influx, is involved in
neuronal death because extracellular Zn2þ can pass through
calcium-permeable channels during synaptic excitation induced
with glutamate. Bell et al. demonstrates a striking pathology-
dependent pattern of glutamatergic synaptic remodeling with
disease progression. Subjects with mild cognitive impairment
display a paradoxical elevation of glutamatergic presynaptic bou-
ton density [76]. Because glutamate excitotoxicity more readily
may occur in elderly people, it is likely that the excitotoxicity is a
cause for age-dependent cognitive decline [77].
Hippocampal CA1 and CA3 are susceptible to neuronal death
induced by glutamate excitotoxicity [78,79] and attention has
been paid to Zn2þ release from hippocampal zincergic neuron
terminals in neurodegeneration. Brief forebrain ischemia in ro-
dents induces selective and delayed neuronal death, particularly
in hippocampal CA1 pyramidal neurons mainly via Zn2þ accu-
mulation [80e82]. Epileptic seizure induces neuronal death in
the hippocampal CA1 and CA3, but not in the dentate gyrus
[83,84]. GluR2-lacking calcium-permeable AMPA receptors may
play a key role for Zn2þ-mediated neurodegeneration [81,85,86].
Therefore, Zn2þ-chelating strategies, which block Zn2þ-mediated
neurodegeneration, may be effective for protecting CA1 and CA3
neurons in neurological disorders associated with glutamate
excitotoxicity. On the other hand, dentate granule cells, which
were scarcely innervated by zincergic neurons, seem to be less
susceptible to Zn2þ neurotoxicity in the hippocampal formation
[87,88]. The levels of messenger RNA encoding the AMPA-
selective glutamate receptor subunit-1, GluR1 and GluR2 are
the highest in the dentate gyrus, followed by the CA1 and CA3
hippocampal subfields [77], suggesting that the dentate gyrus is
susceptible for the influx of Ca2þ and Zn2þ. The GluR1/GluR2
messenger RNA ratios, an index of calcium-permeable AMPA re-
ceptors, are increased in the aged hippocampus, suggesting that
intracellular Zn2þ dyshomeostasis, in addition to intracellular
Ca2þ dyshomeostasis, may contribute to age-related neuronal
death in the hippocampus including the dentate gyrus [77],
which may be linked to cognitive decline in elderly people
(Fig. 1). The social behavioral deficits are accompanied by a
change in AMPA receptor composition, which is regulated by the
brain-enriched microRNA-124, leading to an imbalance between
calcium-permeable and calcium-impermeable AMPA receptors
[89]. The evidence suggests a potential therapeutic avenue for
regulating social behavior in neurodegenerative disorders such as
frontotemporal dementia.
96 A. Takeda, H. Tamano / Archives of Biochemistry and Biophysics 611 (2016) 93e99
Fig. 1. Neuronal death induced by glutamate excitotoxicity often leads to cognitive decline. It is well known that Ca2þ influx through calcium channels (CC) such as NMDA receptors
and calcium-permeable AMPA receptors is involved in postsynaptic neuronal death, while Zn2þ influx at zincergic synapses is also involved in the death.
5. Impact on cognition from synaptic Zn2þ signaling under
diverse synaptic environment
The HPA axis activity, i.e., glucocorticoid secretion, is increased
after exposure to stress and buffers stress. The HPA axis activity is
linked to hippocampal function and also involved in cognitive and
emotional behavior [90]. It is well known that the HPA axis activity
is enhanced in aging [91,92] and neurological disorders [93]. Hip-
pocampal neuronal impairment is associated with the continuous
activation of the HPA axis, which is involved in the occurrence and
progression of cognitive disorders in elderly subjects [94].
Furthermore, correlations have been reported between increases in
HPA axis activity and dementia severity or hippocampal volume
loss in individuals with probable Alzheimer’s disease [95]. Corre-
lations are also reported between increases in HPA axis activity and
depression severity or hippocampal volume loss in human de-
pressives [96].
The hippocampus is a major target of glucocorticoids [97] and
enriched with corticosteroid receptors [98]. The hippocampus is
involved in the negative feedback mechanism in glucocorticoid
secretion. Mineralocorticoid receptors are extensively occupied
with low levels of corticosterone and glucocorticoid receptors are
particularly activated after exposure to stress [90,99]. An increase in
plasma corticosterone level leads to a rapid increase in corticoste-
rone level in the hippocampus, which is in parallel with an increase
in extracellular glutamate level [100]. In the hippocampus,
corticosterone-mediated increase in extracellular glutamate levels
may occur through the action of membrane-associated mineralo-
corticoid receptors and/or glucocorticoid receptors, which in-
creases glutamate release probability in the non-genomic manner
[19,20]. The rapid effects of corticosterone on glutamatergic
neuron activity seem to be linked to the diverse effects on synaptic
plasticity and memory processes in the hippocampus. Corticoste-
rone contributes to increasing the efficacy of glutamatergic trans-
mission by AMPA receptor insertion at synaptic sites through both
the rapid and the delayed (genomic) effects. These effects are of
advantage to the process of synaptic plasticity such as LTP and
cognition [90]. Interestingly, the extracellular zinc is decreased in
the hippocampus in spite of the increase in extracellular glutamate
after exposure to a novel environment [101]. When the decrease is
blocked with CaEDTA, an extracellular Zn2þ chelator, exploratory
activity is simultaneously reduced, suggesting that Zn2þ influx into
hippocampal neurons is linked to exploratory activity [101]. It is
possible that the increase in glucocorticoid secretion induced by
the novelty stress is involved in Zn2þ influx into hippocampal
neurons. Glucocorticoids require intracellular Zn2þ signaling for the
genomic effect [102], suggesting that glucocorticoids modify syn-
aptic Zn2þ signaling under stressful environment and that the
modification is involved in cognitive activity.
In contrast, restraint stress and foot-shock stress increases
glutamate and zinc in the extracellular compartment. Both in-
creases may be mediated by the action of glucocorticoids [103,104].
In amygdala perfusion experiment, psychological stress decreases
zinc in the extracellular fluid with no significant change in extra-
cellular glutamate [104]. The diverse dynamics of Zn2þ and gluta-
mate might be linked to stress-induced glucocorticoid signaling. A
high dose of corticosterone increases Zn2þ release probability from
zincergic neuron terminals through the rapid non-genomic effect
and affects CA1 LTP induction via excess influx of extracellular Zn2þ
into CA1 pyramidal neurons (Fig. 2) [21]. The evidence suggests
that glucocorticoid-mediated excess Zn2þ influx can affect learning
and memory after exposure to stress. It is likely that the increase in
glucocorticoid secretion in neurological disorders is linked to Zn2þ-
mediated cognitive decline. On the other hand, glucocorticoids also
increase Ca2þ influx into hippocampal neurons [105,106].
Glucocorticoid-mediated impairment of CA1 LTP induction is
rescued in the presence of CaEDTA [21], suggesting that the
impairment of LTP induction is due to Zn2þ influx, but not Ca2þ
influx. It has been recognized that excess Ca2þ influx into post-
synaptic neurons induces neuronal death. However, the involve-
ment of excess intracellular Ca2þ signaling in cognitive decline is
poorly understood.
The disease rate of stroke/ischemia is increased with aging. The
concentration of extracellular Kþ is 3.5e5.5 mM under
 A. Takeda, H. Tamano / Archives of Biochemistry and Biophysics 611 (2016) 93e99 97

Fig. 2. Attenuated LTP induction is linked to cognitive decline. Excess Zn2þ influx through calcium channels (CC) such as calcium-permeable AMPA receptors is induced by glu-
tamatergic synapse excitation. The Zn2þ influx subsequently induces cognitive decline via attenuated LTP induction.

physiological condition and reaches approximately 75 mM in brain cognitive decline [120,121]. Metal chaperone, i.e., PBT2, ameliorates
ischemia followed by glutamate excitotoxicity [107]. Glutamate- age-related cognitive decline via increases in dendritic spine den-
mediated neuronal death results in the efflux of intracellular Kþ sity, hippocampal neuron number, and markers of neurogenesis
[108]. Thus it is likely that the increase in extracellular Kþ, which [36], suggesting that the decrease in bioavailable Zn2þ is involved in
non-specifically induces neuronal excitation, leads to Zn2þ-medi- age-related cognitive decline.
ated cognitive decline. Excess glutamatergic neuron activity after In conclusion, extracellular Zn2þ dynamics are diversely modi-
injection of high Kþ into the hippocampal CA1 transiently impairs fied in response to changing synapse environment and involved in
object recognition (Fig. 2) [109]. The impairment is linked to excess cognitive activity in the normal brain. The modification is poten-
Zn2þ release from Schaffer collaterals and the influx of the Zn2þ into tially facilitated with normal aging and critical for cognitive activity
CA1 pyramidal neurons. Even in the dentate gyrus where is scarcely under pathological condition. Extracellular Zn2þ homeostasis is
innervated by zincergic neurons, excess influx of Zn2þ into dentate affected by biologically active substances such as glutamate and
granule cells affects object recognition via attenuated LTP induction glucocorticoids, followed by alteration of intracellular Zn2þ ho-
(Fig. 2) [110]. The findings suggest that the increase in extracellular meostasis (Fig. 3). This process induces Zn2þ-mediated cognitive
Kþ, which induces glutamatergic excitation, occurs in a probability
in the brain of elderly people and is a cause of age-related cognitive
decline.
Numerous studies have demonstrated that traumatic brain
injury and epileptic seizures increase hippocampal neurogenesis in
the rodent and human brain [111]. Newly born dentate granule cells
that arise as a result of traumatic brain injury and epileptic seizures
are integrated into existing hippocampal circuit and may provide
network plasticity for hippocampus-dependent learning and
memory [112,113]. Intracellular Zn2þ signaling is required for hip-
pocampal neurogenesis and has an essential role for hippocampal
neurogenesis after traumatic brain injury and seizure [114e116]. On
the other hand, stress and glucocorticoid strongly inhibit adult
hippocampal neurogenesis [117] and decreased neurogenesis has
been implicated in the pathogenesis of anxiety and depression
[118,119]. Aging is associated with decreased neurogenesis in the Fig. 3. Is Zn2þ-mediated cognitive decline increased along with aging? Glutamatergic
hippocampus, which is linked to hippocampus-dependent synapse excitation and calcium-permeable AMPA receptor expression are increased
with aging as described in the text and critical for Zn2þ influx.
98 A. Takeda, H. Tamano / Archives of Biochemistry and Biophysics 611 (2016) 93e99
decline and may be linked to the changes in Zn2þ-transport protein
expression, e.g., increase in calcium-permeable AMPA receptors. To
understand not only Zn2þ-mediated cognitive decline but also age-
related cognitive decline, it is important to analyze in vivo extra-
cellular Zn2þ dynamics.
6. Perspective
In the last century, cognitive decline in elderly people was
considered as the consequence of neuronal death (Fig. 1). However,
the later analyses suggest that the changes in synaptic function, i.e.,
progressive decrease in neural plasticity, are critical for cognitive
decline and that the changes generally precede loss of neurons in
normal aging (Fig. 2) [122,123]. The changes in synaptic function,
which lead to cognitive decline, may be dynamically linked to
intracellular Zn2þ dyshomeostasis. It is important to analyze Zn2þ-
mediated cognitive decline in the process of aging, based on evi-
dence of the possible involvement of Zn2þ in age-related cognitive
decline [32,36,124] (Fig. 3). A primary important issue that remains
to be addressed is how homeostasis of extracellular and intracel-
lular Zn2þ is controlled in the hippocampus and how it is altered
along with aging.
References
[1] M.Y. Jou, A.G. Hall, A.F. Philipps, S.L. Kelleher, B.J. Lo €nnerdal, Nutrition 139
(2009) 835e841.
[2] J. Dufner-Beattie, Y.M. Kuo, J. Gitschier, G.K. Andrews, J. Biol. Chem. 279
(2004) 49082e49090.
[3] R.J. Cousins, J.P. Liuzzi, L.A. Lichten, J. Biol. Chem. 281 (2006) 24085e24089.
[4] T. Fukada, T. Kambe, Metallomics 3 (2011) 662e674.
[5] C.O. Hershey, L.A. Hershey, A. Varnes, S.D. Vibhakar, P. Lavin, W.H. Strain,
Neurology 33 (1983) 1350e1353.
[6] K. Gellein, J.H. Skogholt, J. Aaseth, G.B. Thoresen, S. Lierhagen, E. Steinnes,
T. Syversen, T.P. Flaten, J. Neurol. Sci. 266 (2008) 70e78.
[7] B. Michalke, V. Nischwitz, Anal. Chim. Acta 682 (2010) 23e36.
[8] J.H. Weiss, S.L. Sensi, J.Y. Koh, Trends Pharmacol. Sci. 21 (2000) 395e401.
[9] W.R. Markesbery, W.D. Ehmann, M. Alauddin, T.I.M. Hossain, Neurobiol.
Aging 5 (1984) 19e28.
[10] A. Takeda, Brain Res. Rev. 34 (2000) 137e148.
[11] A. Takeda, BioMetals 14 (2001) 343e352.
[12] A. Takeda, M. Nakamura, H. Fujii, H. Tamano, Metallomics 5 (2013) 417e423.
[13] C.J. Frederickson, Int. Rev. Neurobiol. 31 (1989) 145e238.
[14] C.J. Frederickson, G. Danscher, Prog. Brain Res. 83 (1990) 71e84.
[15] A. Takeda, H. Tamano, Brain Res. Rev. 62 (2009) 33e34.
[16] A. Takeda, Mol. Neurobiol. 44 (2011) 167e174.
[17] C.B. Sindreu, H. Varoqui, J.D. Erickson, J. Pe rez-Clausell, Cereb. Cortex 13
(2003) 823e829.
[18] A. Takeda, H. Tamano, Significance of the degree of synaptic Zn2þ signaling in
cognition, BioMetals 29 (2016) 177e185.
[19] H. Karst, S. Berger, M. Turiault, F. Tronche, G. Schütz, M. Joe €ls, Proc. Natl.
Acad. Sci. U. S. A. 102 (2005) 19204e19207.
[20] L. Musazzi, M. Milanese, P. Farisello, S. Zappettini, D. Tardito, V.S. Barbiero,
T. Bonifacino, A. Mallei, P. Baldelli, G. Racagni, M. Raiteri, F. Benfenati,
G. Bonanno, M. Popoli, PLoS One 5 (2010) e8566.
[21] A. Takeda, M. Suzuki, H. Tamano, S. Takada, K. Ide, N. Oku, Neurochem. Int. 60
(2012) 394e399.
[22] A. Takeda, H. Tamano, Front. Aging Neurosci. 6 (2014) 26.
[23] A. Takeda, T. Akiyama, J. Sawashita, S. Okada, Brain Res. 640 (1994) 341e344.
[24] A. Takeda, J. Sawashita, S. Okada, Brain Res. 658 (1994) 252e254.
[25] L. Belloni-Olivi, C. Marshall, B. Laal, G.K. Andrews, J. Bressler, J. Neurosci. Res.
87 (2009) 3221e3230.
[26] W. Chowanadisai, S.L. Kelleher, B. Lo €nnerdal, J. Nutr. 135 (2005) 1002e1007.
[27] A.S. Nakashima, R.H. Dyck, Brain Res. Rev. 59 (2009) 347e373.
[28] A. Takeda, A. Minami, S. Takefuta, M. Tochigi, N. Oku, J. Neurosci. Res. 63
(2001) 447e452.
[29] B. Szewczyk, Front. Aging Neurosci. 5 (2013) 33.
[30] S. Sharma, M. Ebadi, Neurochem. Int. 65 (2014) 40e48.
[31] J.R. Nuttall, P.I. Oteiza, Genes Nutr. 9 (2014) 379.
[32] E. Mocchegiani, C. Bertoni-Freddari, F. Marcellini, M. Malavolta, Prog. Neu-
robiol. 75 (2005) 367e390.
[33] T.B. Cole, H.J. Wenzel, K.E. Kafer, P.A. Schwartzkroin, R.D. Palmiter, Proc. Natl.
Acad. Sci. U. S. A. 96 (1999) 1716e1721.
[34] J. Qian, J.L. Noebels, J. Physiol. 566 (2005) 747e758.
[35] P.A. Adlard, J.M. Parncutt, D.I. Finkelstein, A.I. Bush, J. Neurosci. 30 (2010)
1631e1636.
[36] P.A. Adlard, A. Sedjahtera, L. Gunawan, L. Bray, D. Hare, J. Lear, P. Doble,
A.I. Bush, D.I. Finkelstein, R.A. Cherny, Aging Cell. 13 (2014) 351e359.
[37] P.A. Adlard, J. Parncutt, V. Lal, S. James, D. Hare, P. Doble, D.I. Finkelstein,
A.I. Bush, Neurobiol. Dis. 81 (2015) 196e202.
[38] A.I. Bush, J. Alzheimers Dis. 33 (2013) S277eS281.
[39] S. Ayton, P. Lei, A.I. Bush, Neurotherapeutics 12 (2015) 109e120.
[40] D.J. Bobilya, N.A. Gauthier, S. Karki, B.J. Olley, W.K. Thomas, J. Nutr. Biochem.
19 (2008) 129e137.
[41] P. Ballabh, A. Braun, M. Nedergaard, Neurobiol. Dis. 16 (2004) 1e13.
[42] M. Elahy, C. Jackaman, J.C. Mamo, V. Lam, S.S. Dhaliwal, C. Giles, D. Nelson,
R. Takechi, Immun. Ageing 12 (2015) 2.
[43] A. Montagne, S.R. Barnes, M.D. Sweeney, M.R. Halliday, A.P. Sagare, Z. Zhao,
A.W. Toga, R.E. Jacobs, C.Y. Liu, L. Amezcua, M.G. Harrington, H.C. Chui,
M. Law, B.W. Zlokovic, Blood-brain barrier breakdown in the aging human
hippocampus, Neuron 85 (2015) 296e302.
[44] R.A. Yokel, J. Alzheimers Dis. 10 (2006) 223e253.
[45] C.J. Frederickson, L.J. Giblin, A. Krezel, D.J. McAdoo, R.N. Muelle, Y. Zeng,
R.V. Balaji, R. Masalha, R.B. Thompson, C.A. Fierke, J.M. Sarvey,
M. Valdenebro, D.S. Prough, M.H. Zornow, Exp. Neurol. 198 (2006) 285e293.
[46] S. Ueno, M. Tsukamoto, T. Hirano, K. Kikuchi, M.K. Yamada, N. Nishiyama,
T. Nagano, N. Matsuki, Y. Ikegaya, J. Cell Biol. 158 (2002) 215e220.
[47] A. Minami, N. Sakurada, S. Fuke, K. Kikuchi, T. Nagano, N. Oku, A. Takeda,
J. Neurosci. Res. 83 (2006) 167e176.
[48] A. Takeda, H. Tamano, T. Ogawa, S. Takada, M. Ando, N. Oku, M. Watanabe,
Behav. Brain Res. 226 (2012) 259e264.
[49] A. Takeda, H. Tamano, Mol. Neurobiol. (2016, Mar 16).
[50] A. Takeda, M. Suzuki, M. Tempaku, K. Ohashi, H. Tamano, Neuroscience 304
(2015) 209e216.
[51] P. Paoletti, A.M. Vergnano, B. Barbour, M. Casado, Neuroscience 158 (2009)
126e136.
[52] M. Khan, C.R. Goldsmith, Z. Huang, J. Georgiou, T.T. Luyben, J.C. Roder,
S.J. Lippard, K. Okamoto, Proc. Natl. Acad. Sci. U. S. A. 111 (2014) 6786e6791.
[53] S.L. Sensi, L.M.T. Canzoniero, S.P. Yu, H.S. Ying, J.Y. Koh, G.A. Kerchner,
D.W. Choi, J. Neurosci. 15 (1997) 9554e9564.
[54] R.A. Colvin, A.I. Bush, I. Volitakis, C.P. Fontaine, D. Thomas, K. Kikuchi,
W.R. Holmes, Am. J. Physiol. Cell Physiol. 294 (2008) C726eC742.
[55] J.Y. Lee, J.S. Kim, H.R. Byun, R.D. Palmiter, J.Y. Koh, Brain Res. 1418 (2011)
12e22.
[56] Y. Li, C.J. Hough, C.J. Frederickson, J.M. Sarvey, J. Neurosci. 21 (2001)
8015e8025.
[57] E. Pan, X.A. Zhang, Z. Huang, A. Krezel, M. Zhao, C.E. Tinberg, S.J. Lippard,
J.O. McNamara, Neuron 71 (2011) 1116e1126.
[58] J. Emmetsberger, M.M. Mirrione, C. Zhou, M. Fernandez-Monreal,
M.M. Siddiq, K. Ji, S.E. Tsirka, J. Neurosci. 30 (2010) 6538e6547.
[59] S.L. Sensi, D. Ton-That, P.G. Sullivan, E.A. Jonas, K.R. Gee, L.K. Kaczmarek,
J.H. Weiss, Proc. Natl. Acad. Sci. U. S. A. 100 (2003) 6157e6162.
[60] G. Danscher, M. Stoltenberg, J. Histochem. Cytochem 53 (2005) 141e153.
[61] R.A. Colvin, M. Laskowski, C.P. Fontaine, Brain Res. 1085 (2006) 1e10.
[62] C.J. Stork, Y.V. Li, J. Mol. Signal 5 (2010) 5.
[63] A. Takeda, H. Tamano, T. Ogawa, S. Takada, M. Nakamura, H. Fujii, M. Ando,
Hippocampus 24 (2014) 1404e1412.
[64] I. Sekler, A. Moran, M. Hershfinkel, A. Dori, A. Margulis, N. Birenzweig,
Y. Nitzan, W.F. Silverman, J. Comp. Neurol. 447 (2002) 201e209.
[65] C. Nolte, A. Gore, I. Sekler, W. Kresse, M. Hershfinkel, A. Hoffmann,
H. Kettenmann, A. Moran, Glia 48 (2004) 145e155.
[66] P. Aguilar-Alonso, D. Martinez-Fong, N.G. Pazos-Salazar, E. Brambila,
J.A. Gonzalez-Barrios, A. Mejorada, G. Flores, L. Millan-Perezpen ~ a, H. Rubio,
B.A. Leon-Chavez, Brain Res. 1200 (2008) 89e98.

[67] C. Sindreu, A. Baye s, X. Altafaj, J. Pe
rez-Clausell, Zinc transporter-1 concen-
trates at the postsynaptic density of hippocampal synapses, Mol. Brain 7
(2014) 16.
[68] J. Qian, K. Xu, J. Yoo, T.T. Chen, G. Andrews, J.E. Noebels, Knockout of Zn
transporters Zip-1 and Zip-3 attenuates seizure-induced CA1 neuro-
degeneration, J. Neurosci. 31 (2011) 97e104.
[69] S.A. Small, S.A. Schobel, R.B. Buxton, M.P. Witter, C.A. Barnes, Nat. Rev.
Neurosci. 12 (2011) 585e601.
[70] N.C. Danbolt, Prog. Neurobiol. 65 (2001) 1e105.
[71] X.X. Dong, Y. Wang, Z.H. Qin, Acta Pharmacol. Sin. 30 (2009) 379e387.
[72] T.W. Lai, S. Zhang, Y.T. Wang, Excitotoxicity and stroke: identifying novel
targets for neuroprotection, Prog. Neurobiol. 115 (2014) 157e188.
[73] J. Lewerenz, P. Maher, Front. Neurosci. 9 (2015) 469.
[74] C.J. Frederickson, J.Y. Koh, A.I. Bush, Nat. Rev. Neurosci. 6 (2005) 449e462.
[75] S.L. Sensi, P. Paoletti, J.Y. Koh, E. Aizenman, A.I. Bush, M. Hershfinkel, The
neurophysiology and pathology of brain zinc, J. Neurosci. 31 (2011)
16076e16085.
[76] K.F. Bell, D.A. Bennett, A.C. Cuello, Paradoxical upregulation of glutamatergic
presynaptic boutons during mild cognitive impairment, J. Neurosci. 27
(2007) 10810e10817.
[77] S.R. Pagliusi, P. Gerrard, M. Abdallah, D. Talabot, S. Catsicas, Neuroscience 61
(1994) 429e433.
[78] Y.V. Medvedeva, J.H. Weiss, Neurobiol. Dis. 68 (2014) 137e144.
[79] U. Malairaman, K. Dandapani, A. Katyal, PLoS One 9 (2014) e110253.
[80] F. Colbourne, S.Y. Grooms, R.S. Zukin, A.M. Buchan, M.V. Bennett, Proc. Natl.
Acad. Sci. U. S. A. 100 (2003) 2906e2910.
[81] K.M. Noh, H. Yokota, T. Mashiko, P.E. Castillo, R.S. Zukin, M.V. Bennett, Proc.
Natl. Acad. Sci. U. S. A. 102 (2005) 12230e12235.
 A. Takeda, H. Tamano / Archives of Biochemistry and Biophysics 611 (2016) 93e99
[82] C.J. Stork, Y.V. Li, J. Cereb, Blood Flow. Metab. 29 (2009) 1399e1408.
[83] A. Co ^ te
, M. Chiasson, M.R. Peralta 3rd, K. Lafortune, L. Pellegrini, K. To  th,
J. Physiol. 566 (2005) 821e837.
[84] A. Takeda, H. Tamano, A. Nagayoshi, K. Yamada, N. Oku, Neurochem. Int. 47
(2005) 539e544.
[85] S. Liu, L. Lau, J. Wei, D. Zhu, S. Zou, H.S. Sun, Y. Fu, F. Liu, Y. Lu, Neuron 43
(2004) 43e55.
[86] J.H. Weiss, Front. Mol. Neurosci. 4 (2011) 42.
[87] R. Schmidt-Kastner, K.A. Hossmann, Acta Neuropathol. 76 (1988) 411e421.
[88] J.H. Kim, B.G. Jang, B.Y. Choi, L.M. Kwon, M. Sohn, H.K. Song, S.W. Suh, PLoS
One 7 (2012) e48543.
[89] E. Gascon, K. Lynch, H. Ruan, S. Almeida, J.M. Verheyden, W.W. Seeley,
D.W. Dickson, L. Petrucelli, D. Sun, J. Jiao, H. Zhou, M. Jakovcevski,
S. Akbarian, W.D. Yao, F.B. Gao, Nat. Med. 20 (2014) 1444e1451.
[90] C. Sandi, Trends Neurosci. 34 (2011) 165e176.
[91] P.W. Landfield, J.C. Eldridge, Neurobiol. Aging 15 (1994) 579e588.
[92] E. Ferrari, D. Casarotti, B. Muzzoni, N. Albertelli, L. Cravello, M. Fioravanti,
S.B. Solerte, F. Magri, Brain Res. Rev. 37 (2001) 294e300.
[93] N.C. Nicolaides, E. Kyratzi, A. Lamprokostopoulou, G.P. Chrousos,
E. Charmandari, Neuroimmunomodulation 22 (2015) 6e19.
[94] F. Magri, L. Cravello, L. Barili, S. Sarra, W. Cinchetti, F. Salmoiraghi, G. Micale,
E. Ferrari, Stress and dementia: the role of the hypothalamicpituitary-
adrenal axis, Aging Clin. Exp. Res. 18 (2006) 167e170.
[95] J.G. Csernansky, H. Dong, A.M. Fagan, L. Wang, C. Xiong, D.M. Holtzman,
J.C. Morris, Am. J. Psychiatry 163 (2006) 2164e2169.
[96] J.T. O’Brien, D. Ames, I. Schweitzer, P. Colman, P. Desmond, B. Tress, Br. J.
Psychiatry 168 (1996) 679e687.
[97] M. Kawata, K. Yuri, H. Ozawa, M. Nishi, T. Ito, Z. Hu, H. Lu, M. Yoshida,
J. Steroid Biochem. Mol. Biol. 65 (1998) 273e280.
[98] M. Joe €ls, Eur. J. Pharmacol. 583 (2008) 312e321.
[99] M. Joe €ls, H. Karst, R. DeRijk, E.R. de Kloet, Trends Neurosci. 31 (2008) 1e7.
[100] C. Venero, J. Borrell, Eur. J. Neurosci. 11 (1999) 2465e2473.
[101] A. Takeda, N. Sakurada, S. Kanno, A. Minami, N. Oku, J. Neurochem. 99 (2006)
670e676.
[102] A. Takeda, H. Tamano, Metallomics 4 (2012) 614e618.
[103] T. Itoh, T. Saito, M. Fujimura, S. Watanabe, K. Saito, Brain Res. 618 (1993)
318e322.
99
[104] A. Takeda, H. Tamano, S. Imano, N. Oku, Neuroscience 168 (2010) 715e722.
[105] D.S. Kerr, L.W. Campbell, O. Thibault, P.W. Landfield, Proc. Natl. Acad. Sci. U.
S. A. 89 (1992) 8527e8531.
[106] A. Bhargava, O.C. Meijer, M.F. Dallman, D. Pearce, J. Neurosci. 20 (2000)
3129e3138.
[107] A.Z. Hansen, T. Zeuthen, Acta Physiol. Scand. 113 (1981) 437e445.
[108] D. Liu, J.R. Slevin, C. Lu, S.L. Chan, M. Hansson, E. Elme r, M.P. Mattson,
J. Neurochem. 86 (2003) 966e979.
[109] A. Takeda, S. Takada, M. Nakamura, M. Suzuki, H. Tamano, M. Ando, N. Oku,
PLoS One 6 (2011) e28615.
[110] M. Suzuki, Y. Fujise, Y. Tsuchiya, H. Tamano, A. Takeda, Neurochem. Int. 87
(2015) 60e65.
[111] J. Bengzon, Z. Kokaia, E. Elmer, A. Nanobashvili, M. Kokaia, O. Lindvall, Proc.
Natl. Acad. Sci. U. S. A. 94 (1997) 10432e10437.
[112] B.Y. Choi, J.H. Kim, H.J. Kim, B.E. Lee, I.Y. Kim, M. Sohn, S.W. Suh, J. Trace Elem.
Med. Biol. 28 (2014) 474e481.
[113] J.H. Kim, B.G. Jang, B.Y. Choi, L.M. Kwon, M. Sohn, H.K. Song, S.W. Suh, PLoS
One 7 (2012) e48543.
[114] S.W. Suh, S.J. Won, A.M. Hamby, Y. Fan, C.T. Sheline, H. Tamano, A. Takeda,
J. Liu, J. Cereb. Blood Flow. Metab. 29 (2009) 1579e1588.
[115] E.C. Cope, D.R. Morris, S.D. Gower-Winter, N.C. Brownstein, C.W. Levenson,
Exp. Neurol. 279 (2016) 96e103.
[116] J.R. Nuttall, S. Supasai, J. Kha, B.M. Vaeth, G.G. Mackenzie, A.M. Adamo,
P.I. Oteiza, J. Nutr. Biochem. 26 (2015) 1116e1123.
[117] C. Mirescu, E. Gould, Hippocampus 16 (2016) 233e238.
[118] J.S. Snyder, A. Soumier, M. Brewer, J. Pickel, H.A. Cameron, Nature 476 (2011)
458e461.
[119] H.K. Kim, P.V. Nunes, K.C. Oliveira, L.T. Young, B. Lafer, Prog. Neuro-
psychopharmacol. Biol. Psychiatry 67 (2016) 51e57.
[120] E.A. Meyers, K.T. Gobeske, A.M. Bond, J.C. Jarrett, C.Y. Peng, J.A. Kessler,
Neurobiol. Aging 38 (2016) 164e175.
[121] E. Pannese, Brain Struct. Funct. 216 (2011) 85e89.
[122] C. Freitas, F. Farzan, A. Pascual-Leone, Front. Neurosci. 7 (2013) 42.
[123] S. Jinno, J. Chem. Neuroanat. (2015) pii: S0891-0618(15) 00094e0.
[124] M. Warthon-Medina, V.H. Moran, A.L. Stammers, S. Dillon, P. Qualter,
M. Nissensohn, L. Serra-Majem, N.M. Lowe, Eur. J. Clin. Nutr. 69 (2015)
649e661.