*Correspondence: spadilla@well.ox.ac.uk (S.
Padilla-Parra)
The authors’ genetic and pharmacological
approaches stress the importance of the
cortical actin cytoskeleton versus endocy-
tosis in HIV-1 entry, the two processes
being inversely correlated. It has been
shown that the cortical actin network cre-
ates tension that inhibits endocytosis [9].
These new, exciting findings are signifi-
cant in that they allow a better under-
standing of the mechanism of HIV-1
trans-enhancement from DCs to CD4+ T
cells. Interestingly, the multifaceted role of
dynamin in HIV-1 entry seems to extend
beyond trans-enhancement (see [10]). It
will be important to determine dynamin's
function in HIV-1 viral fusion in resting
CD4+ T cells. Several studies show that
the rates of HIV-1 spread from cell to cell
are faster than infection by free virions [11].
It is likely that the role of actin dynamics
and the establishment of the virological
synapse are consequently important in
viral fusion. Moreover, as fusion of the
HIV-1 envelope with the host cell's plasma
membrane is a key step in HIV-1 entry,
conclusive evidence for a mechanism of
HIV-1 fusion and the bona fide role of
DNM2 in all of these contexts is still war-
ranted. Taken together, the work by
Ménager and Littman provides a step for-
ward in helping to elucidate the mecha-
nism of TSPAN7 and DNM2 in HIV-1
trans-infection and may help to reconcile
the reports linking actin dynamics and
dynamin in this process, as both are cru-
cial for HIV-1 viral entry.
Acknowledgments
The S.P.-P. research group is funded by the Nuffield
Department of Medicine Leadership Fellowship from
the University of Oxford. The Wellcome Trust Centre
for Human Genetics is supported by the Wellcome
Trust (Grant No. 090532/Z/09/Z). M.L.D. is supported
by a Wellcome Trust Principal Research Fellowship
(Grant No. 100262/Z/12/Z).
1
Division of Structural Biology, University of Oxford, The
Henry Wellcome Building for Genomic Medicine,
Headington, Oxford OX3 7BN, UK
2
Wellcome Trust Human Genetics, Cellular Imaging Core,
University of Oxford, Oxford, UK
3
The Kennedy Institute of Rheumatology, University of
Oxford, Headington, Oxford OX3 7BN, UK
356 Trends in Molecular Medicine, May 2016, Vol. 22, No. 5
and michael.dustin@kennedy.ox.ac.uk (M.L. Dustin).
http://dx.doi.org/10.1016/j.molmed.2016.03.007
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Spotlight
Acute Glucose
Response Properties
Beyond Feeding
C. Joseph Burnett1,2,3 and
Michael J. Krashes1,2,*
Hypothalamic AgRP neurons po-
tently coordinate feeding behavior
to ensure an organism's viability.
However, their acute role in glu-
cose-regulatory function remains
to be addressed. Steculorum et al.
now report that activation of a spe- To test the contribution of AgRP neurons
cific set of AgRP neurons results in to acute insulin resistance and delineate
an impairment of insulin-stimulated
glucose uptake in brown fat through
a myogenic signature program.
Obesity is one of the most complicated
medical epidemics currently affecting the
Western world. This condition is caused
by a complicated interplay of various fac-
tors, including altered feeding patterns,
decreased energy expenditure, and
altered glucose homeostasis. Many of
these symptoms fall under the control of
neural circuits that maintain homeostatic
equilibrium. For example, agouti-related
peptide (AgRP)-expressing and pro-opio-
melanocortin (POMC) neurons localized in
the arcuate nucleus of the hypothalamus
(ARC) serve as a key substrate in feeding
behavior, both necessary and sufficient to
regulate appropriate energy levels [1–4].
Furthermore, chronic manipulations of
these explicit cell types directly affect glu-
cose homeostasis [5,6]. To date, how-
ever, no studies have investigated the
capacity of these neurons to acutely con-
trol glucose or insulin homeostasis, thus
obscuring direct mechanistic analyses
and the putative neural circuitry involved.
In a new study by Brüning and colleagues
[7] published in Cell, AgRP neurons are
shown to directly affect insulin sensitivity
through perturbation of insulin-stimulated
glucose uptake in brown adipose tissue
(BAT) within minutes to hours (Figure 1).
The researchers utilized molecularly
directed chemogenetic and optogenetic
techniques to stimulate the activity of
either anabolic AgRP, or catabolic POMC
neurons, and analyzed insulin sensitivity in
an acute fashion. Remarkably, activation
of AgRP (but not POMC) neurons rapidly
and robustly impaired both glucose and
insulin tolerance in the absence of food,
exposing a critical regulatory role of AgRP
neurons in modulating peripheral glucose
accumulation during conditions of low
energy resources.
 Brain
Optogenec smulaon

aBNSTvl

ARC

60

45 15

30

laon
cu
Cir Glucose ( )
Insulin ( )

Myostan
S NA transcripts

BAT Mstn
NE

Figure 1. Acute Effects of Agouti-Related Peptide (AgRP) Neuronal Stimulation on Insulin Sensitivity in the Mouse. Chemogenetic or optogenetic
stimulation of AgRP neurons, including their projections to the anterior bed nucleus of the stria terminalis, ventrolateral portion (aBNSTvl) induces changes in blood
glucose occurring within 30 min of stimulation onset. AgRP activity lowers glucose uptake by brown adipose tissue (BAT), rendering these tissues more insulin resistant.
AgRP neurons induce this change by suppressing sympathetic nerve tone (SNA) onto BAT via b-adrenergic-dependent mechanisms. This reduced tone is sufficient to
alter gene expression programs within brown adipocytes to a more myocyte-like expression profile. Importantly, expression of the myocyte-related gene myostatin (Mstn)
is directly linked to acutely suppressing insulin signaling and raising insulin resistance in brown adipocytes. Abbreviations: Arc, arcuate nucleus; NE, norepinephrine.

the peripheral system modulating this
response, the researchers performed
euglycemic/hyperinsulinemic clamps in
mice whereby blood glucose levels are
controlled at an equilibrium point after
acutely raising blood insulin. Compared
with littermate controls, mice with AgRP
neurons active during this clamp demon-
strated a lower glucose infusion rate nec-
essary to maintain constant blood glucose.
This suggested that glucose was not read-
ily cleared from the bloodstream, and was
elevated after AgRP stimulation. Interest-
ingly, this did not impact hepatic glucose
production; instead, insulin-mediated glu-
cose uptake into BAT was drastically dimin-
ished, without affecting white adipose
tissue or skeletal muscle. The authors
found elevated serum insulin, but not glu-
cagon or corticosterone levels after acute
stimulation, further substantiating the

Trends in Molecular Medicine, May 2016, Vol. 22, No. 5 357

capability of AgRP neurons to directly
impair systemic insulin sensitivity. This link
to altered glucose uptake in BAT is
strengthened by similar observations made
in obese mice fed a high-fat diet, which is
known to impact the activity of AgRP [8,9].
If AgRP neuron activity somehow alters
insulin sensitivity, particularly BAT's ability
to take up glucose, BAT itself may undergo
acute genetic expression changes upon
activation of AgRP neurons. To investigate
this, the researchers next looked for
changes in global gene expression in
BAT after AgRP neuron activation using a
microarray-based approach. The most
robustly upregulated RNA products were
transcripts normally produced in muscle
tissue including myostatin (Mstn) (Figure 1);
this upregulation seemed to depend on
feeding state, because fasted mice also
displayed an upregulation of Mstn and
other muscle-associated transcripts.
Exposure to myostatin inhibits differentia-
tion of BAT progenitor cells into brown
adipocytes [10], but b-adrenergic stimula-
tion directs these progenitor cells towards
differentiation into adipocytes. The likely
source of this stimulation is sympathetic
nerve activity (SNA), which may decrease
in response to AgRP activity. Thus, the
investigators directly recorded SNA in
response to AgRP stimulation in anesthe-
tized animals using sharp electrodes to
record from isolated sympathetic nerves
innervating BAT; indeed, this activation
reduced sympathetic nerve tone, presum-
ably lowering b-adrenergic stimulation and
permitting upregulation of Mstn. According
to ex vivo cell culture experiments, myosta-
tin itself was revealed to decrease the ability
of BAT to secrete insulin. To test whether
AgRP activity impacted insulin sensitivity via
b-adrenergic stimulation and myostatin
induction, the authors promoted b-adren-
ergic stimulation and, in a separate series of
358 Trends in Molecular Medicine, May 2016, Vol. 22, No. 5
experiments, blocked myosin activity using
an antibody treatment. Indeed, insulin tol-
erance tests indicated that pretreatment
with a b3-agonist or an anti-myostatin anti-
body blocked the activity of AgRP on insulin
sensitivity.
These experiments address the mecha-
nism of AgRP-mediated systemic insulin
sensitivity, but the neural circuitry relaying
AgRP signaling to the sympathetic ner-
vous system remained unexplored. Thus,
the researchers manipulated specific
AgRP projections to determine which
downstream regions mediated the insulin
sensitivity change observed following
whole-population cell body activation,
which nonspecifically provokes responses
at all efferent targets. Selective stimulation
of AgRP neuron terminals in the anterior
region of the ventrolateral bed nucleus of
the stria terminalis (aBNSTvl), but not the
dorsal medial aBNST, paraventricular
hypothalamus, dorsal raphe, or dorsal
vagal complex, acutely modified insulin
sensitivity through marked induction of
Mstn transcriptional levels in BAT (Fig-
ure 1). Interestingly, while stimulation of
AgRP projections to the lateral hypothala-
mus also altered insulin sensitivity, this
manipulation failed to change Mstn
expression in BAT.
Overall, Steculorum and colleagues have
demonstrated that AgRP neurons exert
more complicated effects on energy bal-
ance than previously understood. These
results suggest that AgRP-mediated
insulin resistance arises quickly and inde-
pendently of its melanocortin receptor-
mediated feeding effects. Specifically,
AgRP activity polysynaptically suppresses
SNA, acutely influencing the genetic com-
position of brown adipocytes to produce
more muscle-related products, especially
myostatin. These exciting findings
advocate obesity interventions targeting
multiple nonoverlapping signaling systems
for an optimal therapeutic benefit. Further
studies mapping the entire wiring diagram
from AgRP!aBNSTvl!BAT will provide
even more pharmacological targeting
opportunities in combating diabetes and
obesity. Clearly, this study provides a
strong bedrock for future investigations
into the interplay between the nervous
and endocrine systems.
1
Diabetes, Endocrinology and Obesity Branch, National
Institutes of Diabetes and Digestive and Kidney Diseases
(NIDDK), National Institutes of Health, Bethesda, MD, USA
2
National Institute on Drug Abuse (NIDA), National
Institutes of Health, Baltimore, MD, USA
3
Brown-NIH Graduate Partnerships Program, Brown
University, Providence, RI, USA
*Correspondence: michael.krashes@nih.gov
(M.J. Krashes).
http://dx.doi.org/10.1016/j.molmed.2016.03.001
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orchestrate feeding behavior rapidly and without training.
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adipose tissue. Cell Published online March 24, 2016.
http://dx.doi.org/10.1016/j.cell.2016.02.044
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