REVIEWS

Brown adipose tissue as a

secretory organ
Francesc Villarroya1,2, Rubén Cereijo2, Joan Villarroya2 and Marta Giralt1,2
Abstract | Brown adipose tissue (BAT) is the main site of adaptive thermogenesis and
experimental studies have associated BAT activity with protection against obesity and metabolic
diseases, such as type 2 diabetes mellitus and dyslipidaemia. Active BAT is present in adult
humans and its activity is impaired in patients with obesity. The ability of BAT to protect against
chronic metabolic disease has traditionally been attributed to its capacity to utilize glucose and
lipids for thermogenesis. However, BAT might also have a secretory role, which could contribute
to the systemic consequences of BAT activity. Several BAT-derived molecules that act in a
paracrine or autocrine manner have been identified. Most of these factors promote hypertrophy
and hyperplasia of BAT, vascularization, innervation and blood flow, processes that are all
associated with BAT recruitment when thermogenic activity is enhanced. Additionally, BAT can
release regulatory molecules that act on other tissues and organs. This secretory capacity of BAT
is thought to be involved in the beneficial effects of BAT transplantation in rodents. Fibroblast
growth factor 21, IL‑6 and neuregulin 4 are among the first BAT-derived endocrine factors to be
identified. In this Review, we discuss the current understanding of the regulatory molecules
(the so‑called brown adipokines or batokines) that are released by BAT that influence systemic
metabolism and convey the beneficial metabolic effects of BAT activation. The identification of
such adipokines might also direct drug discovery approaches for managing obesity and its
associated chronic metabolic diseases.

1
Departament de Bioquímica
i Biomedicina Molecular,
Institut de Biomedicina,
Universitat de Barcelona,
Avda Diagonal 643,
08028‑Barcelona, Catalonia,
Spain.
2
CIBER Fisiopatología de la
Obesidad y Nutrición,
Facultat de Biologia,
Universitat de Barcelona,
Avda Diagonal 643,
08028‑Barcelona, Catalonia,
Spain.
Correspondence to F.V. and
M.G.
fvillarroya@ub.edu; mgiralt@
ub.edu
doi:10.1038/nrendo.2016.136
Published online 12 Sep 2016
Brown adipose tissue (BAT) is the main site of non­
shivering thermogenesis in mammals whereas white
adipose tissue (WAT) is the main depot where meta­bolic
energy is stored, in the form of triglycerides. Brown
adipo­c ytes contain many mitochondria with a high
oxidative capacity and that have uncoupling protein 1
(UCP1) in their inner membrane1. UCP1, which is only
expressed in brown adipocytes, uncouples the respiratory
chain from oxidative phosphorylation yielding a high oxi­
dation rate and enabling the cell to use metabolic energy
to provide heat2. In rat and mouse models, BAT gener­
ates heat to enable the organism to adapt to a cold envi­
ronment and also protects against obesity by promoting
energy expenditure3. Contrary to the prevailing concept
that BAT function is restricted in humans to neonates
and young children, adults have active BAT and activity
of this tissue is systematically reduced in patients with
obesity4. Consistent with its need to adapt to changing
thermal and dietary conditions, BAT is an extremely plas­
tic tissue. When thermogenesis is activated, BAT depots
enlarge via hypertrophic and hyperplasic processes2.
Moreover, sustained thermogenic activation leads to the
‘browning’ of WAT, whereby brown adipocyte-like cells
appear in WAT depots5,6. These brown-like adipocytes,
which are called ‘beige’ or ‘brite’ adipocytes, produce
heat via UCP1‑mediated uncoupling of mitochondrial
respiration, just as that seen in brown adipocytes from
BAT depots7. Some investigators have suggested that the
browning of WAT might protect against obesity8 and
that beige cells might have additional energy expendi­
ture mechanisms that are not mediated by UCP1 (REF. 9).
However, the actual relevance of adipose tissue browning
to systemic metabolism is still a matter of debate10,11.
In the past two decades, a number of WAT-secreted
molecules (called adipokines) have been identified, after
the initial discovery of leptin12. Although the thermo­
genic function of BAT has been recognized by researchers
for many years, the secretory role of this fat depot has
historically been given much less attention than its
other functions. Many of the WAT-secreted adipokines
(including leptin) are poorly expressed in BAT2, which
might have led researchers to mistakenly assume that
BAT only has a limited secretory role. We now know
that BAT effectively secretes low levels of adipokines and

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Key points The secreted protein chemerin has been detected

• The activity of brown adipose tissue (BAT) is associated with protection against
obesity and associated metabolic alterations such as insulin resistance
• Experimental evidence indicates that BAT has systemic effects by secreting
regulatory molecules in addition to its capacity to use metabolic substrates for
thermogenesis
• Brown and beige adipocytes secrete multiple autocrine and paracrine factors that
control expansion and activity of BAT and the extent of browning of white adipose
tissue
• BAT releases endocrine factors that can target peripheral tissues such as white
adipose tissue, liver, pancreas, heart and bone, as well as affect systemic metabolism
by interacting with the CNS
inflammatory cytokines normally seen in WAT2,13. The
specific secretory profile of BAT is quite distinct from that
of WAT, which is perhaps unsurprising considering
that the two tissues have very different, largely oppo­
site, physiological roles in energy metabolism. Here, we
review the data that BAT is a specific source of regulatory
molecules, among them the so‑called brown adipokines
or ‘batokines’. As BAT dissipates metabolic energy as
heat, and actively uses lipids and glucose for oxidation,
molecules secreted by activated BAT might be expected
to support these functions and coordinate BAT activity
with systemic metabolism. Consequently, the identifica­
tion of BAT-secreted molecules, in addition to providing
basic information regarding physiology and metabolism,
might reveal targets for novel drugs to treat or prevent
obesity, type 2 diabetes mellitus and/or dyslipidaemia.
Autocrine and paracrine factors
Many molecules that are released by BAT and/or brown
adipocytes, which might be induced during brown adipo­
cyte differentiation and/or thermogenic, activation have
been identified (FIG. 1; Supplementary information S1
(table)). Although limited direct experimental evidence
exists for the specific actions of these batokines, our
knowledge of their general biological function suggests
that some might have autocrine and/or paracrine roles.
For example, nerve growth factor (NGF) and fibroblast
growth factor 2, are thought to increase sympathetic
innervation and the number of preadipocytes in BAT,
which is relevant to the thermogenic recruitment of
BAT14–16. Moreover, increased innervation17 and hyper­
plasia18 of BAT are required for the enhancement of BAT
thermogenic capacity in physiological conditions of
sustained thermogenic requirement. In addition, the
BAT-mediated secretion of vascular endothelial growth
factor A (VEGFA) can promote vascularization of BAT
itself19,20; increased perfusion is associated with BAT acti­
vation in rodents21 and humans22, especially in response
to cold. BAT-specific overexpression of Vegfa in mice can
increase vascularization and upregulate UCP1, which
thereby increases thermogenesis under chronic cold
exposure23. Conversely, knock out of the Vegfa gene in all
adipose tissue of non-obese mice inhibited the thermo­
genic function of BAT24, which supports the notion that
BAT-secreted VEGFA regulates tissue vascularity. VEGFA
can also target the brown adipocyte itself, which supports
evidence of an additional autocrine role of VEGFA24.
using a signal sequence trap study designed to identify
proteins secreted by brown adipocytes25. Importantly,
cold-induced thermogenic stimulation can repress
chemerin expression in BAT, whereas an obesogenic diet
augmented its expression25. However, the role of BAT-
secreted chemerin is unclear. Chemerin might func­
tion as an autocrine signal via the chemerin receptors
expressed in brown adipocytes to promote lipid accu­
mulation26. Chemerin might also function as a paracrine
agent, and has been reported to chemoattract macro­
phages and other immune cells, such as natural killer
cells and dendritic cells, to adipose tissues27. Although
an endocrine role of BAT-secreted chemerin cannot
be totally ruled out, the lack of correlation between
chemerin expression levels in BAT and circulating levels
of the protein makes this possibility unlikely25.
BAT also secretes nonpeptide signalling molecules.
Several lines of evidence suggest that prostaglandins are
involved in BAT activity and the browning of WAT28,29.
In mice in which the Ptgs2 gene (which encodes cyclo­
oxygenase‑2 and is required for prostaglandin forma­
tion) is disrupted, the browning of WAT is impaired28,
whereas knockout of lipocalin prostaglandin D synthase
impaired BAT activation29. Lipocalin prostaglandin D
synthase is also secreted by BAT and has additional
functions such as transport of thyroid hormones and
vitamin  A derivatives in the blood30. Moreover, in
another study, prostaglandin E2 was found to directly
induce browning in WAT31. Specifically, pharmacologi­
cal inhibition or small interfering RNA-mediated down­
regulation of microsomal prostaglandin E synthase 1, a
cyclooxygenase‑2 downstream synthase responsible
for the biosynthesis of prostaglandin E, repressed the
capacity of mouse preadipocytes to differentiate into
beige adipocytes31. Cold-induced thermogenic activa­
tion of BAT also induces the expression of nitric oxide
synthase in this adipose depot, which might contrib­
ute to the enhanced blood flow that is essential for the
trophic adaptation of BAT to increased thermogenesis32.
Moreover, inorganic nitrate can activate the browning
of WAT by inducing the synthesis of nitric oxide33. The
treatment of rats with nitrate promoted WAT brown­
ing, and exposure of mouse preadipocyte cell cultures
to nitrate can induce the acquisition of a beige adipo­
cyte phenotype33. These effects have been proposed to
be driven by nitric oxide generated from nitrate via serial
reduction to nitrite and then to nitric oxide33. The nucle­
otide adenosine is also released by brown adipocytes and
BAT stimulated by the sympathetic nervous system; in
turn, this secretion can enhance thermogenic activation
in an autocrine manner34.
Thermogenic activation of BAT is also associated
with a strong enhancement of intracellular lipolysis,
which produces fatty acids that fuel intracellular ther­
mogenesis2. Fatty acids might be released from brown
adipo­cytes to mediate relevant signalling processes either
locally or at a distance. For example, the noradrenergic-
mediated stimulation of lipolysis in cultured brown
adipocytes can trigger the release of fatty acids to
the medium2, but if this effect is simply an in vitro

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REVIEWS

endocannabinoid system37. Peripheral antagonists tar­

Endothelial cell
NO VEGFA
• ANGPTL8
• BMP8b
• Endothelin-1
• FGF21
Eosinophil
• IL-6
• LPGDS Metrnl
• sLR11
• Slit2-C
• Adenosine
• Prostaglandins
• Endocannabinoids
Adiponectin
Brown/beige
adipocyte
• NRG4
• NGF TH2 cytokines
Neuron
M2 macrophage
Figure 1 | The autocrine and paracrine factors released NaturebyReviews
brown and beige
| Endocrinology
adipocytes. Brown and beige adipocytes secrete factors with autocrine actions that
lead to enhanced (by BMP8b, endothelin‑1, FGF21, IL‑6, LPGDS) or inhibited (by sLR11)
thermogenic activity. Other factors secreted by brown and/or beige adipocytes act
locally on other cell types. Nerve growth fibre (NGF) secreted by brown adipocytes
promotes sympathetic innervation, secreted vascular endothelial growth factor A
(VEGFA) targets endothelial cells to induce vascularization of BAT and nitric oxide (NO)
increases blood flow, processes that are associated with brown adipose tissue (BAT)
recruitment in response to sustained thermogenic activation. Meteorin-like protein
(Metrnl) is secreted by beige, and possibly brown, adipocytes and promotes eosinophil
activation, which results in the recruitment of alternatively activated M2 macrophages.
The release of adiponectin by beige adipocytes also seems to induce alternative
activation and recruitment of M2 macrophages. ANGPTL8, angiopoietin-like 8; BMP,
bone morphogenetic protein; LPGDS, lipocalin D synthase; NRG4, neuregulin 4; sLR11,
soluble LDL receptor 11.
phenomenon or if activation of BAT in vivo can induce
a release of fatty acids is still unclear. Moreover, whether
endogenous oxidation can account for most of the
fatty acids released via the lipolysis of the intracellu­
lar tri­glyceride stores is also still to be determined.
Nevertheless, fatty acids released by BAT under lipolytic
stimulation might still act as signalling molecules with
autocrine and/or paracrine actions. For example, the
established action of free fatty acids released by lipolysis
on the activation of the UCP1‑mediated uncoupling of
mitochondria35 is c­ onsistent with this possibility.
Notably, of the factors that are released by BAT that
are thought to have autocrine or paracrine actions, most
have positive effects on thermogenic activation and
BAT recruitment. However, the soluble form of the LDL
receptor, sLR11, suppresses thermogenesis in brown
adipocytes, despite being increased by the cold-induced
activation in BAT36. This function might occur via an
autocrine-based homeostatic mechanism and is thought
to prevent excessive energy wastage upon activation of
thermogenesis36. An analogous scenario seems to occur
for endocannabinoids. The activation of BAT and the
browning of WAT via cold exposure or β3‑adrenergic
stimulation are associated with the induction of the
geting the cannabinoid receptor‑1 activate adaptive
thermo­genesis in BAT38, and the cold-mediated increase
in the endocannabinoid tone has, therefore, been pro­
posed as part of an autocrine negative feedback mech­
anism that controls β3‑adrenoceptor-induced BAT
activation and WAT browning37. Finally, endothelin‑1,
which is released by brown adipocytes, can also repress the
thermogenic activities of brown and beige adipose tissue39.
Bone morphogenetic proteins
Bone morphogenetic proteins (BMPs) have strong
effects on the processes leading to the differentiation
of brown and beige adipocytes40–42 (FIG. 2). BMPs, which
belong to the transforming growth factor β (TGFβ)
superfamily of extracellular signalling proteins, are
multifunctional regulators of development and tissue
homeostasis43. Several BMPs regulate adipogenic pre­
cursor cell commitment and differentiation: BMP2 and
BMP4 have been primarily associated with white adipo­
cyte differentiation40,43, and BMP7 seems to, at least in
part, regulate brown adipo­genesis41. Conversely, TGFβ
and activin A can inhibit the differentiation of both
white and brown adipocytes44. Most BMP signalling is
mediated via serine-threonine kinase receptor type 1
(BMPR1) and/or type 2 (BMPR2) and downstream
activation of the SMAD transcription factors40,43. In
humans, adipose tissue distribution and obesity have
been associated with changes in adipose tissue expres­
sion and/or serum levels of various BMPs and adipocyte-
specific expression levels of genetic variants of BMPR1A
and BMPR2 (REF. 45).
BMP7, which is expressed early in brown adipo­
genesis, is necessary for the formation of classic BAT
depots. Bmp7 knockout mice have a marked decrease in
the total mass of BAT (but not WAT), and adenoviral-
mediated overexpression of Bmp7 in adult mice is report­
edly associated with BAT recruitment, increased energy
expenditure and weight loss41. Moreover, specific dele­
tion of Bmpr1a in Myf5‑expressing cells (which give rise
to classic brown adipocytes) results in a severe paucity of
BAT and the compensatory activation of WAT browning46.
BMP7 also regulates beige adipogenesis and promotes
the commitment of mouse and human WAT-derived pro­
genitor cells into the beige adipocyte lineage47. However,
adenovirus-mediated overexpression of Bmp7 did not
induce WAT browning in adult mice41, while in another
study, BMP7 and a β3‑adrenergic receptor agonist
synergistically induced the browning of WAT47.
Within adipose tissues, BMP7 is mainly produced by
stromal vascular cells47. Mechanistically, autocrine and/or
paracrine BMP7 signalling in resident adipocyte precur­
sors induces the expression of PRDM16 and PGC1a41,47,
which are crucial to drive the differentiation of adipocyte
precursors to a brown adipocyte fate48. BMP7 enhances
thermogenic gene expression (including that of UCP1)
and mitochondrial biogenesis during brown adipocyte
differentiation, and increases mitochondrial activity
and fatty acid catabolism in mature brown adipocytes49.
BMP7 also acts on the central nervous system to regulate
thermogenesis50.

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obesity54. The overexpression of BMP4 alone, and/or the

Precursor cell
Brown Beige
preadipocyte preadipocyte
Commitment
BMP7 BMP4
Differentiation GDF8 Differentiation
Brown
Follistatin Follistatin
adipocyte
Beige
adipocyte
BMP8b
Endocrine?
Figure 2 | BMPs in brown and beige adipogenesis and function.
Nature ReviewsBMP7 is produced
| Endocrinology
by brown adipose tissue stromal vascular cells and promotes brown adipocyte
commitment and differentiation. By contrast, BMP4 is induced in adipogenic precursor
cells in white adipocytes where, in addition to promoting white adipogenesis
(not shown), it regulates beige precursor cell commitment and differentiation in the
presence of thermogenic activation stimuli. Growth/differentiation factor 8 (GDF8) is
produced by brown adipocytes in response to hunger-related stimuli in the central
nervous system and seems to have negative effects on brown and beige adipocyte
thermogenic activity. Follistatin is secreted by brown, and possibly beige, cells and
induces their activity by repressing GDF8 signalling. BMP8b is secreted by mature brown
adipocytes and it enhances the responsiveness of brown fat to sympathetic activation in
autocrine and endocrine manners. BMP, bone morphogenetic protein.
Whereas BMP7 seems to selectively induce brown
adipo­genesis, BMP4 was historically thought to be a
specific regulator of white adipogenesis51. However,
BMP4 not only acts as an endogenous regulator of white
adipocyte cell commitment and differentiation, it also
promotes beige adipocyte differentiation42. In mice, adi­
pose-tissue-specific overexpression of Bmp4 can promote
the browning of subcutaneous WAT, increase energy
expenditure and improve insulin sensitivity42. Conversely,
adipose-tissue-specific Bmp4‑null mice had enlarged
white adipocytes and developed insulin resistance42. In
vitro, BMP4 treatment of mesenchymal progenitors and
both white and brown preadipocytes can induce BAT-like
differentiation in mouse models52 and primary human adi­
pose stem cells53. Moreover, BMP4 is induced and secreted
from preadipocytes during preadipocyte differentiation51.
By contrast, BMP7 is poorly expressed in stromal cells
isolated from human WAT, and BMP2 is downregulated
during adipogenesis54. Consequently, the autocrine and/or
paracrine function of BMP4 seems to induce both white
and beige adipogenic commitment and differentiation.
This process is counterbalanced by the secretion of the
BMP4/BMP7 inhibitor Gremlin 1 from preadipocytes and
adipocytes, which is notably upregulated in hypertrophic
silencing of Gremlin 1, increases the transcriptional acti­
vation of PPARγ and drives preadipocytes towards an oxi­
dative beige adipose phenotype54. Consequently, BMP4
secreted by white adipocytes is an integral feedback regu­
lator for the adipogenic commitment and differentiation
of both white and beige adipocytes. BMP4 and BMP7
have been proposed to be useful for modulation of the
brown and beige adipocyte differentiation processes in
relation to the treatment of obesity and its comorbidities55.
However, this potential use of BMP modulation should be
considered with caution given the possible adverse effects,
including ectopic bone formation56.
BMP8b is mainly produced by mature brown adipo­
cytes and BMP8B expression is increased in response
to thermogenic and nutritional factors, such as cold
exposure or a high-fat diet57. BMP8b functions locally
by enhancing the response of BAT to β3‑adrenergic
stimulation57 and signals in BAT via SMAD1, SMAD5
and SMAD8 and increased activation of p38‑MAPK57;
however, the membrane receptor complex mediating
this response has not yet been identified. Expression of
Bmp8b in BAT is higher in female mice than in male
mice, and can be induced by oestrogens58. The BMP8b
mechanism might, therefore, contribute to sex-specific
differences in BAT activity. Beyond its local effects
in BAT, BMP8b also functions in the hypothalamus
to increase activation of BAT thermogenesis by the
­sympathetic nervous system57.
The expression and secretion of other BMP/TGFβ
family members in brown and/or beige adipocytes and
their precursors might also modulate adipocyte func­
tion. For example, both growth/differentiation factor 5
(GDF5)59 and BMP9 (REF. 60) can control brown and
beige adipogenesis and enhance energy expenditure in
mice. Conversely, inhibition of TGFβ signalling pro­
motes the browning of WAT and protects mice from
diet-induced obesity61,62. Furthermore, upon stimu­
lation of hunger-related neural circuits, BAT activates
a skeletal-muscle-like gene expression programme
that includes the induction of growth/differentiation
factor 8 (GDF8; commonly known as myostatin), a
secreted myokine that inhibits BAT thermogenesis,
browning of WAT and metabolic activity63,64. Further
research is needed to confirm that GDF8 is a negative
autocrine factor in the regulation of BAT. Conversely,
follistatin, which is a soluble glycoprotein that is
expressed in BAT exposed to cold temperatures, might
have a positive autocrine function on brown adipocytes
by directly antagonizing the effects of GDF8 (REFS 65,66).
Immunometabolism and BAT secretory role
In obese states, the production of proinflammatory
cytokines in WAT is dramatically increased due to a
quantitatively higher contribution by white adipocytes
and a greater number of infiltrating proinflammatory
immune cells (mainly classically activated macro­
phages) than in lean states 67. This upregulation of
proinflammatory cytokines in WAT is thought to be a
main cause of the onset of obesity-associated insulin
resistance67. By contrast, the role of immune cells in

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BAT and cytokine-mediated crosstalk between brown
adipocytes, preadipocytes and immune cells has only
begun to be explored in the past few years. In BAT, the
transcript levels of proinflammatory cytokines are gen­
erally lower than in WAT, which probably reflects the
enhanced anti-inflammatory phenotype of the resident
immune cells in BAT13,68. However, increased levels of
expression of some proinflammatory cytokines, such as
tumour necrosis factor (TNF), C‑C motif chemokine 2
(also known as monocyte chemoattractant protein 1)
and IL‑1, have been reported in BAT under obesogenic
conditions in rodents2,69,70; these molecules have also
been proposed to counteract the increase in thermo­
genesis in these tissues71–73. As the synthesis of these
cytokines is exacerbated under conditions of obesity,
evaluating the relative contribution of BAT (and spe­
cifically of brown adipocytes) to increased circulating
levels of cytokines and systemic insulin resistance has
been challenging. Conversely, some cytokines previously
regarded as proinflammatory do not seem to have this
specific role in BAT physiology and a complex scenario
is beginning to emerge. For example, the expression
and release of IL‑1α and IL‑6 from brown adipocytes is
induced in BAT in response to thermogenic activation74.
Conversely, GDF5 is expressed and released by BAT
under proinflammatory stimuli but can promote
BAT activation and the browning of WAT depots59,75.
The synthesis of anti-inflammatory cytokines in BAT
has also been associated with heat production. Immune
cells located in healthy adipose tissues have emerged as
key functional regulators of insulin sensitivity in brown
and white adipocytes and thermogenic adaptation
to cold stress in brown adipocytes13,68. Alternatively-
activated macrophages, under the coordination of type 2
T helper (TH2) response immune cells contribute to the
recruitment and/or activation of BAT and the browning
of WAT; however, this mechanism is currently poorly
understood68,76–78, and might involve the direct produc­
tion of catecholamines68. To orchestrate this immuno­
logical component of thermogenic activation, a complex
paracrine network of cytokines (such as, IL‑4, IL‑13,
IL‑5 and IL‑33) and TH2 immune cells (alternatively-
activated M2 macrophages, eosinophils and type 2
innate lymphoid cells) is established in BAT upon cold
stimulation68,77–79. Moreover, direct interactions of IL‑4
and IL‑13 in beige precursor cells have been proposed
to promote their differentiation into thermogenic adipo­
cytes76. Although these TH2 cytokines are believed to be
released mostly (but not exclusively) by diverse immune
cell populations, brown adipocytes can also communi­
cate with immune cells through the release of certain fac­
tors. For example, meteorin-like protein (Metrnl), which
is produced by brown and beige adipocytes in response
to cold stimuli, can activate eosinophils to produce IL‑4
and thereby activate macrophages to the alternatively
activated M2 phenotype80. Moreover, beige-adipocyte-
derived adiponectin can contribute to the proliferation
of alternatively activated M2 macrophages to ensure
long-term browning of WAT depots81. These findings
highlight a specific role for adiponectin secreted from
beige cells, which was previously considered to be pri­
marily a WAT adipokine. Further research is needed to
clarify the roles of the cytokines and chemokines that
are secreted by brown and beige adipocytes compared
with those produced by BAT-infiltrating immune cells.

Endocrine factors secreted by BAT

Brown/beige Evidence from transplantation studies. In addition to
adipocyte the molecules that seem to have autocrine and/or para­
crine functions, BAT might also be a source of factors
that have an endocrine role. In the late 1980s, T3 was first
recognized as a BAT-released hormone82. This assertion
IGFBP2 was based on the finding that in brown adipocytes, but
• FGF21 not white adipocytes, noradrenergically-regulated type 2
• FGF21 • NRG4 • IL-6
• FGF21 iodothyronine deiodinase activity was present that con­
• IL-6 • IGF1
• Slit2-C
• BMP8b verts T4 to T3 (REF. 83). T3 can induce the expression of
• IL-6 UCP1 and other components of the thermogenic BAT
machinery 84 and in enhanced thermogenic activa­
Bone White adipose Brain Liver Pancreas Heart
tion by cold, BAT contributes to the systemic levels
Figure 3 | Putative batokines and target organs. EndocrineNaturefactors released
Reviews by brown
| Endocrinology of T3 (REFS 82,85).
and beige adipocytes might signal to distinct tissues, including the brain. Some of these The concept that BAT might be a source of secreted
factors (fibroblast growth factor 21 (FGF21), Slit2‑C) might also target white adipose regulatory molecules has gained particular attention in
tissue (WAT) in an endocrine (and autocrine and/or paracrine) mechanism to induce the light of research exploring the effect of BAT transplan­
browning of WAT and the metabolic processes associated with provision of substrates to tation on systemic metabolism and other physiological
fuel thermogenesis. In the pancreas, FGF21, IL‑6 and possibly angiopoietin-like 8 functions (FIG. 3; TABLE 1). The transplantation of embry­
(ANGPTL8) might improve insulin secretion and β‑cell function. Brown adipose tissue onic BAT into diabetic adult mice can significantly
(BAT) might also release NRG4 to attenuate lipogenesis in the liver, insulin-like growth improve glycaemic conditions, WAT inflammation
factor binding protein 2 (IGFBP2) to promote bone formation and FGF21 and IL‑6 to
status and systemic adipokine profiles (for example, by
increase cardiac substrate oxidation and protect the heart from hypertrophy and
oxidative stress. The endocrine factors released by BAT might modulate systemic upregulating adiponectin)86,87. The antidiabetic action
metabolism indirectly through the central nervous system, and BAT-released factors such of transplanted BAT has been proposed to be due to its
as FGF21, IL‑6 and possibly BMP8b might influence sympathetic nervous system activity release of insulin-like growth factor 1 (IGF1)86. Other
and other processes such as feeding, circadian behaviour and female endocrine function. investigators have used adult mouse BAT for trans­
IGF1, insulin-like growth factor 1; NRG4, neuregulin 4. plantation into diabetic mice and found similar results,

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Table 1 | The effects of experimental BAT transplantation

Source of BAT Recipient of BAT Main outcomes Refs
Embryonic mice Streptozotocin-induced Improved glycaemia 86
(E16.5–17.5) diabetic mice
Embryonic mice Autoimmune-mediated Improved glycaemia 87
(E16.5–17.5) type 1 diabetic mice
Adult mouse iBAT HFD-induced obese mice Improved glucose homeostasis; decreased body weight 88
Adult mouse iBAT HFD-induced obese mice Improved glucose homeostasis; decreased body weight 89
Adult mouse iBAT HFD-induced obese mice Increased energy expenditure; increased sympathetic 91
activity
WAT from exercised Adult control mice Improved glucose homeostasis; increased glucose 92
mice (beige-enriched) uptake in skeletal muscle
Adult mouse iBAT HFD-induced obese mice Improved glucose homeostasis; cardioprotection 94
Adult mouse iBAT ob/ob mice Increased energy expenditure; improved glucose 90
tolerance; reduced hepatic steatosis
Adult rat iBAT Polycystic ovary syndrome Improved insulin sensitivity; improved fertility 95
model in adult rats
Human beige cells Glucose intolerant NSG Improved glucose tolerance 93
differentiated in vitro mice
E, embryonic day; HFD, high-fat diet; iBAT, interscapular brown adipose tissue; NSG, NOD scid gamma; WAT, white adipose tissue.

including a reversal of high-fat-diet-induced obesity,
improved glucose homeostasis and enhanced insulin
sensitivity88,89. BAT transplantation also had beneficial
effects in the ob/ob mouse model of obesity by reducing
body weight while increasing energy expenditure and
levels of adiponectin90. In transplantation studies, grafted
BAT tends to involute and lose the high intrinsic meta­
bolic and thermogenic activities that would be expected
to explain the beneficial effects of the BAT transplants88,89.
As a result, the benefits of BAT transplantation have
been hypothesized as being caused by the release of reg­
ulatory factors from the transplanted tissue. Moreover,
in some studies, BAT transplantation can promote whole
body sympathetic nervous system activity and elevated
sympathetic tone to heart, liver, muscle, visceral WAT
and BAT from recipient mice, which suggest that puta­
tive BAT-released factors might have effects on the
CNS91. Moreover, the transplantation of subcutaneous
WAT from mice exposed to exercise (which induces
browning in WAT) improves glucose homeostasis,
whereas transplantation of WAT from sedentary mice
does not92. This exercise-induced enrichment of beige
adipocytes in WAT might in turn enhance the release
of putative beige-derived adipokines (as discussed later
in the text). The implantation of in vitro-differentiated
human beige adipocytes to non-obese diabetic NOD-
scid IL2rgnull mice, which are glucose intolerant, fed
a high-fat diet can also improve glucose tolerance93.
Moreover, BAT transplantation has potential clinical
benefits beyond improvements in energy homeostasis
and glycaemia, such as protecting against experimen­
tally induced myocardial infarction in mice94. Notably,
these effects were lost when Ucp1‑null (that is, thermo­
genically inactive) BAT was used for transplantation94.
Finally, BAT transplantation has also been reported to
improve glucose homeostasis in the dehyroepiandroster­
one-induced rat model of polycystic ovary syndrome95.
Surprisingly, BAT transplantation was found to reverse
anovulation, hyperandrogenism and polycystic ovaries,
as well as improve fertility in the rat polycystic ovary
syndrome model, which might indicate the existence
of yet unidentified BAT-derived factors involved in the
control of the female reproductive system95.
Fibroblast growth factor 21. The secreted factor fibro­
blast growth factor 21 (FGF21) promotes glucose use in
adipose tissues and improves glycaemia and lipidaemia96.
In most conditions, the liver is the main source of sys­
temic FGF2197; however, when BAT is thermogenically
activated, brown adipocytes express and release high
amounts of FGF21 (REFS 98,99). Expression of FGF21
and its release from BAT is regulated by noradrenergic,
cAMP-mediated mechanisms, the same intracellular
pathways that induce thermogenic gene expression99.
An autocrine role for FGF21 is also likely, as FGF21
itself promotes thermogenesis and metabolite oxidation
in BAT100,101. However, activated BAT seems to contrib­
ute to systemic FGF21 levels. For example, in mice, cold
exposure leads to increased plasma levels of FGF21,
which is associated with increased FGF21 release by BAT
and repressed Fgf21 expression in the liver99. Moreover,
in Ucp1‑null mice, a dramatic rise in Fgf21 expression in
BAT accompanied by a large increase in serum levels of
FGF21 but unaltered hepatic FGF21 gene expression is
seen102,103. Expression and release of FGF21 are induced
in skeletal muscle as a consequence of mitochondrial
DNA mutations or experimentally-induced alterations
in mitochondrial function104–106. Genetic inactivation of
the mitochondrial protein UCP1 might lead to similar
mitochondrial dysfunction in BAT, which might explain
enhanced FGF21 expression and release in BAT from
Ucp1‑null mice102,103. In any case, these observations are
highly suggestive that a rise in BAT FGF21 expression is
capable of influencing systemic FGF21 levels. High levels

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of FGF21 are produced by activated BAT, which is con­
sistent with several metabolically healthy effects known to
be concomitantly associated with BAT activity and with
FGF21 action: prevention of hyperglycaemia and hyper­
lipidaemia and protection against obesity via enhance­
ment of energy expenditure96. Furthermore, the finding
that FGF21 might exert part of its metabolic effects
independently of UCP1‑mediated thermogenesis103,107
highlights that BAT can influence metabolism beyond
brown adipocyte thermogenic activity itself.
IL‑6. Thermogenic, noradrenergic-mediated activation
of brown adipocytes is associated with enhanced IL‑6
expression74. This observation might seem inconsistent
with the role of IL‑6 as a proinflammatory cytokine and
the inverse association between inflammatory signalling
and BAT activation71–73. However, IL‑6 is now considered
distinct from standard proinflammatory cytokines and
is known to be released by skeletal muscle in response to
exercise and to promote insulin sensitivity108. In adipose
tissue, IL‑6‑mediated signalling promotes M2 macro­
phage activation by sensitizing these cells to the action
of IL‑4, which in this context is concomitant with an
enhancement of insulin sensitivity109. Investigators have
tried to identify the adipokines mediating the effect of
BAT transplantation using tissue transferred from IL‑6-
null mice89. In this study, the lack of IL‑6 expression
impaired the beneficial effects of BAT transplantation
on metabolic health89. However, this manipulation also
attenuated the increase in FGF21 levels that are normally
seen after transplantation of wild-type BAT, so the role of
IL‑6 as the main mediator of beneficial BAT transplan­
tation effects remains unclear. Finally, IL‑6 is required
for the induction of browning of WAT in response to a
cold environment, as shown by the blunted induction of
UCP1 protein in subcutaneous WAT after cold exposure
of Il‑6-null mice110.
Neuregulin 4. Secreted by brown adipocytes, Neure­
gulin 4 (NRG4), is also thought to promote neurite
outgrowth111. The Nrg4 gene has been found in tran­
scriptomic microarray assays among genes predicted to
encode secreted proteins that are induced during brown
adipocyte differentiation and enriched in BAT111,112.
Gain‑of‑function and loss‑of‑function studies in mice
demonstrated that Nrg4 acts on the liver to attenuate
hepatic lipogenic signalling112.
Insulin-like growth factor-binding protein 2. BAT might
also influence bone. Beige adipocytes have been proposed
to secrete insulin-like growth factor-binding protein 2
and Wnt10b, which stimulate bone formation and bone
turnover with a net effect of bone gain113. These factors
might function at a distance and locally, as some bone
marrow adipose cells might acquire beige fat features and
influence osteogenesis via secretion of these factors113.
Retinol binding protein 4. The vitamin A transporter,
Retinol binding protein 4 (RBP4), has also been pro­
posed to be an adipokine released by WAT that pro­
motes insulin resistance114. Brown adipocytes under
nora­drenergic activation also release RBP4, but whether
this factor favours the transport of retinoids released
after lipolysis of retinyl esters in response to noradr­
energic activation of BAT, or is associated with RBP4
functions unrelated to retinoid transport is unclear115.
However, BAT-released RBP4 might not be associated
with insulin resistance given that cold-induced activation
of BAT is associated with insulin sensitization116.
Angiopoietin-like 8. The expression of Angiopoietin-
like 8 (Angptl8), also commonly called lipasin, RIFL
(‘refeeding induced fat and liver’) or betatrophin, is
induced in BAT in response to cold117. At present, the
biological significance of enhanced Angptl8 release by
activated BAT is unclear. Angptl8 can repress the activ­
ity of lipoprotein lipase118, which might counteract the
increase in lipoprotein lipase activity in cold-induced
BAT that supports thermogenesis2. Angptl8 was also
thought to enhance pancreatic β‑cell replication119.
Given the association between high BAT activity and
improved glucose homeostasis and insulin sensitivity,
BAT was thought to signal to β cells via Angptl8; how­
ever, new studies have questioned the role of Angptl8
in β-cell expansion120, which has led investigators to
dismiss this scenario.
Other factors. In a study of the YY1 transcription factor
in BAT, a number of other factors were found to be
secreted by brown adipocytes in vitro, including growth/
differentiation factor 15, angiopoietin-like 6, neurome­
din B and nesfatin121. These factors are known to act
as hormones and have been associated with energy
expenditure; however, further research is needed for
a direct demonstration of the role of BAT in releasing
physiologically relevant amounts of these factors that can
signal other tissues and organs121.
Beige versus brown adipocyte secretion
Is the secretory profile of beige adipocyte different
from that of classic brown adipocytes? At present, we
have no clear answer to this question and the evidence
is indirect. However, research in this field has given
some compelling evidence that beige cells can confer
protection against obesity and associated metabolic
disturbances. Moreover, these cells might have the
ability to secrete regulatory molecules that mediate
these effects. The use of non-biased approaches, such
as transcriptomic microarray assays, to identify differ­
entially or preferentially expressed genes in beige ver­
sus brown adipocytes has not found genes that encode
secretory factors6,122,123. However, analysis of gene tran­
scripts in cell cultures revealed that FGF21 is prefer­
entially expressed in beige adipocytes compared with
classic brown adipocytes6,122. As highlighted earlier in
the text, subcutaneous WAT (but not BAT) expresses
large amounts of adiponectin in response to cold,
which promotes M2 macrophage proliferation and
subsequent browning of WAT81. Although the details
of this process have not yet been fully established, this
finding might reflect a particularly relevant adiponec­
tin secretory capacity of beige cells relative to classic

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Box 1 | Future research directions
• Specific animal models are needed to investigate the
effect of tissue-specific knock out of putative
adipokines in brown adipocytes to further advance
knowledge of the secretory function of brown adipose
tissue
• Investigations need to compare the secretory function
of beige adipocytes with that of classic brown
adipocytes given their involvement in the browning of
white adipose tissue
• A thorough analysis of the secretome of brown and
beige adipocytes is needed to facilitate the translation
of mouse and rat studies to human applications
beige cell-enriched adipose tissue from patients with
pheochromocytoma compared with pure WAT (R.
Cereijo & F. Villarroya, unpublished work), which
suggests that this putative preferential brown and/or
beige adipokine identified in rodent studies is relevant
in humans. In another report, implantation of mice with
human beige cells can improve glucose homeostasis93,
which further supports the notion that human beige
cells can secrete factors that have systemic actions sim­
ilar to those seen in rodents. However, human BAT and
beige fat depots might not be identical to those in mice.
For example, according to their gene expression profile,
human supraclavicular BAT depots might in fact consist
of both classic brown adipocytes and beige adipocytes130.

brown adipocytes. Similarly, Metrnl is preferentially, Conclusions

but not exclusively, expressed in beige versus brown
adipocytes80.
Evidence of human batokines
Specific information about the BAT secretome in
humans is still scarce. This paucity of data is possibly
due to the intrinsic difficulties of conducting human
studies and the awareness of the presence of active BAT
in human adults. Moreover, inter-species differences in
secreted capacities should not be underestimated.
In some studies, investigators have reported preferen­
tial expression of type 2 iodothyronine deiodinase, an
enzyme that converts T4 to T3, in human BAT versus
WAT124, which suggests that human BAT is likely to
produce T3 from T4 as in rodent BAT. However, the
adipose-tissue-specific differences between humans and
rodents are highlighted by the case of FGF21, one of the
few factors secreted by BAT for which some informa­
tion in humans is available. In contrast to mice and rats,
expression of FGF21 is practically undetectable in human
WAT125,126; however, expression of FGF21 in human
BAT is significant, regardless of whether it is composed
mainly of classic brown adipocytes or beige adipocytes127.
Moreover, in vitro human brown adipocytes derived
from fetal BAT express and secrete FGF21 (REF. 127).
Cold exposure in humans leads to increases in plasma
levels of FGF21 (REF. 128), and the data from several stud­
ies has indicated that plasma levels of FGF21 reflect BAT
activity in humans128,129. However, further research will
be needed to assess the actual capacity of human BAT to
be a source of systemic FGF21 in humans. In an RNAseq
analysis, NRG4 was found to be highly expressed in
The standing association between BAT activity and
protection against obesity, hyperglycaemia and hyper­
lipidaemia cannot be explained only by the ability of
this tissue to burn glucose and lipids for thermogen­
esis; however, its capacity to signal other organs and
regulate systemic metabolism might also contribute to
this mechanism. BAT transplantation might even be a
strategy to take advantage of the beneficial effects of
brown adipocytes131. However, the major goal for future
research in this field is the precise identification of the
brown adipokines or batokines that are candidates
for drug development to combat metabolic diseases.
Research advances in the coming years are expected
to refine the analysis of the secretome in brown and/or
beige adipocytes (BOX 1). For example, in a study using
non-biased proteomics Slit2‑C, a cleaved form of the
extracellular protein Slit was identified as a secreted
factor that promotes energy expenditure and glucose
homeostasis, and seems to function in an autocrine
and/or endocrine manner 132. Moreover, the bio­
informatic prediction of how well a candidate protein
is secreted using -omics analysis of differential gene
expression in BAT has yielded promising results.
Ultimately, experimental models of gene manipula­
tion leading to BAT-specific gain and loss‑of‑function
approaches might be especially useful to establish the
actual systemic role of putative brown adipokines. In
the coming years, research should focus on identify­
ing and characterizing new adipokines from BAT, and
establishing their role in metabolic regulation and if
they can be pharmacologically exploited to treat obesity
or ­metabolic diseases.

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Acknowledgements
The authors wish to acknowledge the support of grants
SAF2014‑55725 from the Ministerio de Ciencia e Innovación
(MINECO) and PI14/00063 from the Instituto de Salud
Carlos III, Spain, co‑financed by the European Regional
Development Fund (ERDF), and the European Community
Seventh Framework Program (FP7 BetaBat).
Author contributions
The authors contributed equally to all aspects of the prepa-
ration of this article.
Competing interests
The authors declare no competing interests.
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