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Endothelial GLUT1
deficiency

Glucose
LRP1
A

Reduced brain
glucose uptake

Microvascular
rarefaction

BBB leakage

Energy deficit

Hypoperfusion

Altered homeostasis

Reduced A
clearance

Amyloid pathology

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2015 Nature America, Inc. All rights reserved.

Synaptic dysfunction and neurodegeneration

COMPETING FINANCIAL INTERESTS

The author declares no competing financial interests.

Cognitive
deficits

Figure 1 Mechanisms of brain dysfunction and damage caused by GLUT1 deficiency. Endothelial
GLUT1 deficiency leads to reduced brain glucose transport, vascular rarefaction and disruption of
the BBB, as well as reduced A clearance by suppressing vascular LRP1 expression. These events
result in energy deficit, reduced cerebral blood flow (hypoperfusion), altered homeostasis of the
brain microenvironment and enhanced amyloid pathology. The resulting synaptic dysfunction and
neurodegeneration in turn lead to cognitive deficits.

early studies of cerebral metabolism1. A similar

situation is found in patients with SLC2A1 deficiency, a rare genetic disease associated with
prominent neurological deficits, in whom a
ketogenic diet is highly beneficial14.
Could GLUT1 be used to develop new therapeutic interventions in AD? If a reduction in
endothelial GLUT1 enhances AD pathology,
it is conceivable that restoring GLUT1 levels
could ameliorate brain dysfunction and damage
in AD. To begin to address this question,

Little is known about the mechanism causing GLUT1 dysregulation in AD. GLUT1
expression is controlled by hypoxia-inducible
factor 1 (HIF1,). Given that HIF1 is downregulated in AD15, it is conceivable that HIF1
suppression leads to reduced GLUT1 expression. However, earlier studies have indicated
that SLC2A1 mRNA is not reduced in AD,
implicating post-translational mechanisms8.
Thus, further studies on the molecular bases of
GLUT1 reduction are needed to provide some
indication of how to counteract it.
Irrespective of the many questions outstanding, the data of Winkler et al.4 demonstrate a
multifaceted role of glucose transport in the
maintenance of brain structure and function, and
unveil a damaging interaction with AD pathology. This may open new therapeutic avenues for
this devastating neurodegenerative disease.

Winkler et al.4 performed adenoviral gene transfer in APPSw mice deficient in Slc2a1. They found
that restoration of GLUT1 in the hippocampus
greatly reduced local A levels. Similar results
were obtained with viral gene transfer of LRP1,
the A vascular transport protein suppressed by
GLUT1 deficiency. Although the authors did not
demonstrate rescue of neuronal function and
behavior, the findings provide proof of principle
that upregulation of GLUT1 clears the brain of
amyloid and could have beneficial effects.

1. Hoyer, S., Oesterreich, K. & Wagner, O. J. Neurol. 235,

143148 (1988).
2. Harik, S.I., Kalaria, R.N., Andersson, L., Lundahl, P. &
Perry, G. J. Neurosci. 10, 38623872 (1990).
3. Mamelak, M. J. Alzheimers Dis. 31, 459474 (2012).
4. Winkler, E.A. et al. Nat. Neurosci. 18, 521530
(2015).
5. Nordberg, A., Rinne, J.O., Kadir, A. & Lngstrm, B.
Nat. Rev. Neurol. 6, 7887 (2010).
6. Jagust, W.J. et al. J. Cereb. Blood Flow Metab. 11,
323330 (1991).
7. Kalaria, R.N. & Harik, S. J. Neurochem. 53,
10831088 (1989).
8. Mooradian, A.D., Chung, H.C. & Shah, G.N. Neurobiol.
Aging 18, 469474 (1997).
9. Jack, C.R. et al. Lancet Neurol. 12, 207216 (2013).
10. Bateman, R.J. et al. N. Engl. J. Med. 367, 795804
(2012).
11. de le Monte, S.M. & Tong, M. Biochem. Pharmacol.
88, 548559 (2014).
12. Krikorian, R. et al. Neurobiol. Aging 33,
425.e19425.e27 (2012).
13. Reger, M.A. et al. Neurobiol. Aging 25, 311314 (2004).
14. Pearson, T.S., Akman, C., Hinton, V.J., Engelstad, K. &
De Vivo, D.C. Curr. Neurol. Neurosci. Rep. 13, 342
(2013).
15. Liu, Y., Liu, F., Iqbal, K., Grundke-Iqbal, I. &
Gong, C.-X. FEBS Lett. 582, 359364 (2008).

Cocaine shapes chromatin landscapes via Tet1

Anne E West
Chronic cocaine exposure induces long-lasting, transcription-dependent changes in neuronal function. A genome-wide
sequencing study shows how cocaine changes the epigenome to exert specific, long-lasting effects on neuronal transcription.
Memories are the essence of a life. Ask yourself Who am I? and you trigger a mental
Anne E. West is in the Department of Neurobiology,
Duke University Medical Center, Durham, North
Carolina, USA.
e-mail: west@neuro.duke.edu

478

movie of schoolrooms, a wedding kiss, a

tasty French pastry or a sled. Neuroscientists
have long sought to understand how past
experiences are encoded in the brain.
Recently, the field has fallen in love with
the idea of epigenetics, in which the brain
plasticity narrative of dynamic learning

and persistent memory finds a physical

instantiation in the biochemical modifications of genomic DNA and its associated
histone proteins. Methylation of DNA is
strongly associated with persistent biological processes in cells, such as X chromosome
inactivation and gene imprinting. When

volume 18 | number 4 | april 2015 nature neuroscience

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2015 Nature America, Inc. All rights reserved.

Marina Corral Spence/Nature Publishing Group

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H3K27Ac
5hmC

AAAA

AAAA

Cocaine
H3K4me3
Tet1

H3K4me1

5hmC

Figure 1 Cocaine-dependent changes in the architecture of the 5hmC landscape are associated
with changes in gene expression. In the NAc, 5hmC is enriched at active gene enhancers, across
gene bodies, and near exon boundaries. Enhancers are indicated by peaks of histone H3 modified
by acetylation at lysine 27 (H3K27Ac) and by monomethylation at lysine 4 (H3K4me1). Promoters
are indicated by peaks of histone H3 modified by trimethylation at lysine 4 (H3K4me3) and
transcription starts sites are shown by the arrows. Cocaine exposure reduces Tet1 expression
and results in reduced (shown) or increased 5hmC deposition at both enhancers and splice
sites. Changes in 5hmC at exon boundaries (*) are associated with changes in splice site usage.
Alternative splicing of an mRNA that generates transcript A-B-C-D before cocaine generates
transcript A-B-D after cocaine.

it was discovered that DNA methylation

can be dynamically lost through the action
of a family of enzymes called the Tets, this
suggested a mechanism by which environmental experience could be remembered
via its effect on epigenetic chromatin regulation, gene expression and neuronal function on a behaviorally relevant timescale.
Now, in this issue of Nature Neuroscience,
Feng et al.1 provide new evidence for the
biological functions of Tets in brain plasticity, reporting that Tet1 is a target of regulation by cocaine and that decreases in Tet1
remodel the chromatin landscape in ways
that change the expression of functionally
important neuronal genes.
In mammalian cells, the methylation of
cytosines (5mC) in genomic DNA is mediated
by a small family of DNA methyltransferases.
Cytosine methylation has been predominantly
studied in the context of CpG dinucleotides,
although in the brain non-CpG methylation
seems likely to be important as well2. In 2009,
Kriaucionis and Heintz3 made the intriguing
discovery that DNA from Purkinje cell nuclei
contained an unusual DNA nucleotide, which
they identified as 5-hydroxymethylcytosine
(5hmC). In parallel, Tahiliani et al.4 identified 5hmC in embryonic stem cells and
demonstrated that the human TET1 enzyme
catalyzed the conversion of 5mC to 5hmC. In
addition to Tet1, Tet2 and Tet3 were found to
have similar enzymatic activities, and knockout and knockdown studies quickly confirmed
the requirement for these enzymes in the generation of 5hmC during cellular differentiation
and embryonic development57.
The brain contains some of the highest
levels of 5hmC in the body, and they increase
from early postnatal development through
adulthood, suggesting a role for 5hmC in
mature brain function8. Consistent with

this possibility, Tet1 knockout mice show

diminished expression of activity-regulated
genes, abnormal hippocampal long-term
depression and impaired memory extinction, although the connection between these
phenotypes and environmentally driven
changes in Tet1 function or 5hmC distributions remains unknown9. Furthermore,
Tet3 expression is enhanced in infralimbic
cortex following fear conditioning and Tet3
knockdown in this brain region is associated
with impaired fear conditioning, further
suggesting a link between Tets, 5hmC
and transcription-dependent behavioral
plasticity10.
Repeated cocaine exposure can lead to
addiction, which is one of the most persistent forms of environmentally induced and
transcription-dependent brain plasticity.
Thus, Feng et al.1 asked whether cocaine
regulates the Tets in the nucleus accumbens (NAc), a brain region required for the
rewarding effects of cocaine. Mice that had
been repeatedly exposed to cocaine had significantly less Tet1 mRNA and protein in the
NAc than controls, whereas levels of Tet2 and
Tet3 were unchanged. TET1 mRNA expression was also reduced in the NAc of brains
from humans addicted to cocaine, suggesting
the relevance of this regulatory pathway for
addiction. Knocking down the expression of
Tet1 in the NAc enhanced cocaine-induced
conditioned place preference, a behavioral
assay of the rewarding effects of cocaine. By
contrast, overexpression of Tet1 in the NAc
impaired conditioned place preference.
These data show that the cocaine-dependent
decrease in Tet1 expression is required for
behavioral plasticity induced by repeated
exposure to cocaine, and, overall, these data
indicate that Tet1 functions to negatively
regulate cocaine reward.

nature neuroscience volume 18 | number 4 | april 2015

Despite the reduced expression of Tet1

in the NAc after cocaine, the authors found
no effect of cocaine on the global levels of
either 5hmC or 5mC as a percentage of total
cytosine in the NAc. However, when they
performed genome-wide sequencing for the
distribution of 5hmC, they found that cocaine
induced significant changes (both increases
and decreases) in 5hmC levels at more than
11,000 sites across the genome, distributed
both in intergenic regions and across gene
bodies. These data are consistent with a
model in which local recruitment of Tet1 to
specific gene regulatory elements mediates
the regulation of a discrete set of target genes.
To test this model, the authors compared the
cocaine-induced changes that they observed
in 5hmC levels at different kinds of genomic
elements with alterations in the expression of
nearby genes.
In the intergenic regions, 5hmC was
enriched at elements marked by acetylation of histone H3 on Lys27 and monomethylation on Lys4, which are the histone
modifications most strongly associated
with active distal gene enhancers. Even
though cocaine decreases Tet1 expression
and Tet1 promotes the conversion of 5mC
to 5hmC, cocaine exposure was associated
with an equal number of enhancers showing increases in 5hmC compared to those
showing decreases in 5hmC. The loss of Tet1
was sufficient to mediate increases in 5hmC:
when the authors knocked down Tet1 in the
NAc in the absence of cocaine exposure, they
observed enhanced 5hmC at several loci that
also showed cocaine-induced enhancement.
The mechanism by which loss of Tet1 leads
to increased 5hmC was not resolved in this
study, but the authors speculate that it could
arise from secondary dysregulation of 5hmC
metabolism, non-enzymatic contributions of
the Tets to chromatin regulation11, or functional compensation by Tet2 and/or Tet3.
Regardless, as the authors did not detect a
correlation between cocaine-induced 5hmC
changes at enhancers and global changes in
gene expression profiled by RNA-seq, the
functional importance of these chromatin
changes remains unknown.
Over gene bodies, 5hmC was depleted
at transcriptional start sites and enriched
both upstream of transcription ending sites
and in regions flanking exon boundaries,
all of which are consistent with previous
reports8,12. Following cocaine, the authors
found that changes in 5hmC were enriched
near splice sites. Furthermore, by comparing
5hmC distributions at exon boundaries with
RNA-seq data, they found an intriguing correlation between changes in 5hmC levels and
479

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2015 Nature America, Inc. All rights reserved.

news and views

the usage of specific splice sites. Specifically,
alternative splice isoforms upregulated after
cocaine were more likely to be associated
with increased 5hmC at the corresponding
splice site, whereas 5hmC at splice sites
was more likely to be reduced for isoforms
downregulated after cocaine (Fig. 1).
These data provide functional evidence
that 5hmC may regulate splice site usage,
which will be an important area for future
investigation.
The last question the authors asked is
whether changes in 5hmC contribute to the
persistent changes in NAc physiology that
are both induced by chronic cocaine and
relevant to addiction. The authors found
a significant global correlation between
genes that showed increased 5hmC following repeated cocaine and those that showed
enhanced steady-state expression 24 h after
withdrawal from chronic cocaine. The correlation with increased 5hmC was even stronger for genes that were induced by a cocaine
challenge. These data therefore indicate

that 5hmC levels not only reflect current

transcriptional states, but also predict the
potential for genes to turn on in response to
a future stimulus. Finally, the authors demonstrated that, at least for a subset of genes,
both mRNA induction and cocaine-induced
changes in 5hmC can persist for at least 1
month after the cessation of cocaine exposure. Thus, rather than being just an intermediate in the demethylation of DNA, these
data support a model of 5hmC as a meaningful epigenetic mark of its own, with potential
functions in the maintenance of transcriptional memory.
This work by Feng et al.1 underscores the
importance of epigenetic mechanisms of chromatin regulation in the long-lasting changes
in neuronal gene expression that are induced
by chronic cocaine. Furthermore, their findings demonstrate the power of genome-level
sequencing techniques to open new windows
of understanding into the mechanisms of
neuronal adaptation. The challenge for the
future will be to distill the detailed chromatin

landscape revealed here into a set of principles

for gene regulation that will better link
molecular mechanism via cellular function to
the maladaptive circuit changes that underlie
drug addiction.
COMPETING FINANCIAL INTERESTS
The author declares no competing financial interests.
1. Feng, J. et al. Nat. Neurosci. 18, 536544
(2015).
2. Lister, R. et al. Science 341, 1237905 (2013).
3. Kriaucionis, S. & Heintz, N. Science 324, 929930
(2009).
4. Tahiliani, M. et al. Science 324, 930935
(2009).
5. Koh, K.P. et al. Cell Stem Cell 8, 200213 (2011).
6. Ito, S. et al. Nature 466, 11291133 (2010).
7. Dawlaty, M.M. et al. Dev. Cell 24, 310323
(2013).
8. Szulwach, K.E. et al. Nat. Neurosci. 14, 16071616
(2011).
9. Rudenko, A. et al. Neuron 79, 11091122
(2013).
10. Li, X. et al. Proc. Natl. Acad. Sci. USA 111,
71207125 (2014).
11. Kaas, G.A. et al. Neuron 79, 10861093
(2013).
12. Wen, L. et al. Genome Biol. 15, R49 (2014).

Carrot or stick in motor learning

Dagmar Sternad & Konrad Paul Krding
A study shows that reward and punishment have distinct influences on motor adaptation. Punishing mistakes
accelerates adaptation, whereas rewarding good behavior improves retention.
We both love salsa dancing, but learning
salsa is not easy. When one partner misses
a step, the other may punish him with a
frown, but when he masters a new move, her
praise rewards himor does it make him
complacent? Carrot or stick: the manner
by which reward and punishment affects
motor learning is a long-standing question
in education, sports, therapy and beyond.
In this issue of Nature Neuroscience, Galea
et al.1 address this question using a simple reaching task in a perturbed visual
environment.

Dagmar Sternad is in the Departments of Biology,

Electrical and Computer Engineering, and Physics,
and the Center for the Interdisciplinary Research on
Complex Systems, Northeastern University, Boston,
Massachusetts, USA, and Konrad Paul Krding
is in the Sensory Motor Performance Program,
Rehabilitation Institute of Chicago, Chicago, Illinois,
USA, and the Departments of Physical Medicine
and Rehabilitation, and Physiology, Northwestern
University, Chicago, Illinois, USA.
e-mail: dagmar@neu.edu or kk@northwestern.edu

480

The authors build on previous studies that

have shown the crucial importance of reward
on retention2,3, but they now contrast the
effect of reward with that of punishment by
differentiating their effects on acquisition rate
and retention. This study examined participants moving a cursor to targets displayed on
a screen, steering with their hand movements
hidden from view. To create a learning challenge, the cursor position was rotated by 30
degrees and participants had to practice to
successfully reach the target. This exercise is
similar to moving a computer mouse when you
turn it upside down: a challenge that one masters with practice. The specific question of this
paper was how money received for good performance (reward) or lost for bad performance
(punishment) would affect the rate of learning
and the retention of the acquired performance.
The authors found that punishment accelerated the rate of adaptation, whereas reward
improved retention of the new mapping.
This study is at the crossroads of at least
three research disciplines that have examined the consequences of motivational and
error feedback on motor performance:

neuroscience, computational science and, of

course, psychology (Fig. 1). These fields have
approached this issue in different ways and
each can inform and motivate future directions in motor control.
In psychology, reward and punishment
have long been recognized as instrumental for
learning. As early as 1898, Edward Thorndikes
law of effect stated that if a response leads to a
satisfying state of affairs it will be strengthened and, conversely, if it leads to unpleasant
consequences it will be weakened4. The thesis
that reward is a better motivator than punishment was also at the core of B.F. Skinners
principle of reinforcement. Operant conditioning developed systematic reinforcement
schedules to enhance learning and thereby
shape behavior5. However, social psychologists
have also reminded us that human nature is
far more nuanced and more than a collection
of systematically reinforced associations. Many
studies have highlighted the mediating effects
of emotions, such as threat, anxiety, pride or
shame, on behavior. Invoking stereotypes, such
as inferior performance of females in mathematics or athletics, lowers test performance.

volume 18 | number 4 | april 2015 nature neuroscience