J Physiol 603.5 (2025) pp 1091–1107 1091
The Jou nal o Physiology
In acellula signalling in a e ial chemo ecep o s du ing
acu e hypoxia and glucose dep i a ion: ole o ATP
Ma ía To es-López1,2 , Pa icia González-Rod íguez1,2,3 , Olalla Colinas1,2,3 ,Hee-SoolRho
1,
Ho ensia To es-To elo1,2 ,An onioCas ellano
1,2 ,LinGao
1,2,3, Pa icia O ega-Sáenz1,2,3
and José López-Ba neo1,2,3
1Ins i u o de Biomedicina de Se illa (IBiS), Hospi al Uni e si a io Vi gen del Rocío/CSIC/Uni e sidad de Se illa, Se illa, Spain
2Depa amen o de Fisiología Médica y Bio ísica, Facul ad de Medicina, Uni e sidad de Se illa, Se illa, Spain
3Cen o de In es igación Biomédica en Red sob e En e medades Neu odegene a i as (CIBERNED)
Handling Edi o s: Ha old Schul z & And ew Holmes
The pee e iew his o y is a ailable in he Suppo ing In o ma ion sec ion o his a icle
(h ps://doi.o g/10.1113/JP287130#suppo -in o ma ion-sec ion)
Abs ac igu e legend Schema ic ep esen a ion o he dis inc mechanisms in ol ed in acu e oxygen and glucose
sensing by ca o id body glomus cells.
M. To es-López and P. González-Rod íguez con ibu ed equally o his wo k
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y. DOI: 10.1113/JP287130
This is an open access a icle unde he e ms o he C ea i e Commons A ibu ion License, which pe mi s use, dis ibu ion
and ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed.
1092 M. To es-López and o he s J Physiol 603.5
Abs ac The ca o id body (CB) is he main oxygen (O2) sensing o gan ha media es e lex hype -
en ila ion and inc eased ca diac ou pu in esponse o hypoxaemia. Acu e O2sensing is an in insic
p ope y o CB glomus cells, which con ain special mi ochond ia o gene a e signalling molecules
(NADH and H2O2) ha modula ememb aneK
+channels in esponse o lowe ed O2 ension
(hypoxia). In pa allel wi h hese memb ane-associa ed e en s, glomus cells a e highly sensi i e o
mi ochond ialelec on anspo chain (ETC)inhibi o s.I wassugges ed ha adec easeinoxida i e
p oduc ion o ATP is a c i ical e en media ing hypoxia-induced cell depola iza ion. He e, we show
ha o enone [an inhibi o o mi ochond ial complex (MC) I] ac i a es a and mouse glomus cells
bu abolishes hei esponsi eness o hypoxia. Ro enone does no p e en u he ac i a ion o he
cells by cyanide (a blocke o MCIV) o glucose dep i a ion. Responsi eness o glucose dep i a ion
is enhanced in O2-insens i e glomus cells wi h gene ic dis up ion o MCI. These indings sugges
ha acu e O2sensing equi es a unc ional MCI bu ha a dec ease in in acellula ATP, p esumably
p oduced by he simul aneous inhibi ion o MCI and MCIV, is no in ol ed in hypoxia signalling.
In suppo o his concep , ATP le els in single glomus cells we e unal e ed by hypoxia, bu apidly
declined ollowingexposu eo hecells olowglucoseo oinhibi o so oxida i ephospho yla ion.
These obse a ions indica e ha a educ ion in in acellula ATP does no pa icipa e in physio-
logical acu e O2sensing. Howe e , local dec eases in ATP o glycoly ic o igin may con ibu e o low
glucose signalling in glomus cells.
(Recei ed 18 June 2024; accep ed a e e ision 27 Janua y 2025; i s published online 11 Feb ua y 2025)
Co esponding au ho s J. López-Ba neo and P. O ega-Sáenz: Ins i u o de Biomedicina de Se illa (IBiS), Hospi al
Uni e si a io Vi gen del Rocío/CSIC/Uni e sidad de Se illa, Se illa, Spain. Email: [email p o ec ed] and go [email p o ec ed]
Key poin s
The ca o id body con ains oxygen-sensi i e glomus cells wi h specialized mi ochond ia ha
gene a e signalling molecules (NADH and H2O2) oinhibi memb aneK
+channels in esponse
o hypoxia.
Glomus cells a e highly sensi i e o elec on anspo chain (ETC) blocke s. I was sugges ed
ha a dec ease in in acellula ATP is he main signal inducing K+channel inhibi ion and
depola iza ion in esponse o hypoxia o ETC blockade.
Ro enone,aninhibi o o mi ochond ial complex(MC)I, ac i a esglomuscellsbu abolishes hei
esponsi eness o hypoxia. Howe e , o enone doesno p e en u he ac i a iono glomuscells
by cyanide (an MCIV blocke ) o glucose dep i a ion.
Single-cell ATP le els we e unal e ed by hypoxia, bu dec eased apidly ollowing exposu e o
glomus cells o 0 mM glucose o inhibi o s o oxida i e phospho yla ion.
A educ ion in in acellula ATP does no pa icipa e in signalling acu e hypoxia. Howe e , i may
con ibu e o hypoglycaemia signalling in glomus cells.
0Ma ía To es-López go he Deg ee in Biochemis y and Mas e ’s in Physiology and Neu oscience a he
Uni e si y o Se ille. She mo ed o he Ins i u e o Biomedical Resea ch o Ba celona o s udy axonal
neu odegene a ion in p od omal Pa kinson’s disease. F om 2020, she joined he Ins i u e o Biomedicine
o Se ille o wo k on he molecula mechanism unde lying O2sensing by ca o id body glomus cells as
pa o he PhD p ojec . Pa icia Gonzalez-Rod iguez is a neu oscien is and physiologis whose esea ch
ocuses on oxygen sensing, mi ochond ial dys unc ion and Pa kinson’s disease. D González-Rod íguez
ob ained he deg ee in Biology and comple ed he PhD a he Uni e si y o Se ille, Spain. He pos doc o al
aining has been conduc ed a ou esea ch ins i u ions ac oss h ee coun ies: EPFL in Swi ze land,
No hwes e n Uni e si y in Chicago (USA), he Uni e si y o Minneso a Medical Cen e in Minneapolis
(USA) and he Uni e si y o Se ille in Spain.
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
J Physiol 603.5 In acellula signalling in a e ial chemo ecep o s 1093
In oduc ion
The ca o id body (CB) is he main a e ial chemo-
ecep o and p o o ypical acu e oxygen (O2)sensing
o gan in mammals ha is esponsible o he hypoxic
en ila o y esponse (HVR). I is composed o clus e s
o glomus cells ha a e inne a ed by senso y ne e
ib esanda einclosecon ac wi hcapilla ies( o
de ailed e iews see I u iaga e al., 2021; O ega-Sáenz
& López-Ba neo, 2020). Glomus cells a e elec ically
exci able and elease neu o ansmi e s in esponse o
lowe ed O2 ension (hypoxia), he eby ac i a ing a e en
ib es o he pe osal ganglion synapsing on b ain s em
espi a o y cen es (see López-Ba neo, 2022). Hypoxia
inhibi s he ac i i y o backg ound and ol age-dependen
K+channels in glomus cells (Buckle , 1997; Delpiano
& Heschele , 1989; Kim e al., 2009; Lopez-Ba neo
e al., 1988; López-López e al., 1989; Pee s, 1990;
Pé ez-Ga cía e al., 2004; S ea & Nu se, 1991), leading o
memb ane depola iza ion, ex acellula Ca2+in lux and
neu o ansmi e sec e ion (Buckle & Vaughan-Jones,
1994; López-Ba neo e al., 1993; U eña e al., 1994).
In addi ion o hese memb ane-associa ed e en s, i is
known ha CBs a e s ongly ac i a ed by mi ochond ial
inhibi o s ( o e e ences see Mulligan & Lahi i, 1981;
Wilson e al., 1994) and ha se e al mi ochond ial
pa ame e s (e.g. memb ane po en ial o NADH le els)
in dissocia ed glomus cells a e modula ed by changes in
O2 ension (Buckle & Tu ne , 2013; Duchen & Biscoe,
1992a, b). Simila o hypoxia, he ac i a ion o glomus
cells (inc eased cy osolic Ca2+o sec e o y ac i i y) by
mi ochond ial elec on anspo chain (ETC) blocke s
equi es an in lux o ex acellula Ca2+, indica ing
ha mi ochond ia can signal memb ane ion channels
(O ega-Sáenz e al., 2003; Wya & Buckle , 2004).
O e he pas ew yea s, s udies on gene ically modi ied
mouse models ha e p o ided s ong e idence o suppo
a comp ehensi e mi ochond ial- o-memb ane signalling
(MMS) pa hway o acu e O2sensing by glomus cells,
which combines memb ane and mi ochond ial esponses
o hypoxia. CB glomus cells cons i u i ely exp ess high
le els o HIF2α(Gao e al., 2017; Tian e al., 1998), which
induces he exp ession o a ypical mi ochond ial complex
(MC) IV subuni iso o ms (Kadenbach & Hu emann,
2015) con e ing upon mi ochond ia a high sensi i i y
o changes in O2 ension (see Bishop & Ra cli e, 2020;
Colinas e al., 2023; Mo eno-Domínguez e al., 2020).
Hypoxia dec eases MCIV ac i i y and causes a backlog
o elec ons along he ETC, esul ing in an inc ease in
he educed s a us o MCIII and in he a io o educed
coenzyme Q (QH2) o coenzyme Q (Q). These dynamic
changes slow down MCI NADH dehyd ogenase ac i i y
and esul inanaccumula iono NADHandaninc eased
p oduc ion o eac i e oxygen species (ROS), gene a ed
in MCIII and MCI and con e ed o H2O2by supe -
oxide dismu ases. NADH and H2O2a e mi ochond ial
signals ha a e equilib a ionwi h hecy osolmodula e
ion channel unc ion (A ias-Mayenco e al., 2018;
Fe nández-Agüe a e al., 2015; Jiménez-Gómez e al.,
2023; Mo eno-Domínguez e al., 2020). An unsol ed
mechanis ic ques ion ega ding he MMS model o
acu e O2sensing is whe he o no hypoxia causes a
dec ease in cy osolic ATP le els in glomus cells ( o a
de ailed discussion, see López-Ba neo & O ega-Sáenz,
2022). A p io i, he ole o a d op in in acellula ATP in
signalling physiological acu e hypoxia is coun e in ui i e
since ATP is needed o suppo essen ial unc ions in
ac i e cells, such as main enance o ionic g adien s, Ca2+
channel unc ion and neu osec e ion. Howe e , i was
epo ed ha TASK-like channels, which media e he
backg ound K+cu en inhibi ed by hypoxia in glomus
cells, a e ac i a ed by in acellula ATP (Va as e al.,
2007). The e o e, i was sugges ed ha a dec ease in
cy osolic ATP du ing hypoxia is he signal esponsible
o he hypoxia-induced dec ease in K+conduc ance and
cell depola iza ion (Wya & Buckle , 2004). Challenging
his p oposal, i was ound ha CB glomus cells om
mice lacking TASK1 and TASK3 channel subuni s ha
o m ATP-ac i a ed backg ound K+channels (Kim
e al., 2009) ha e no mal sec e o y esponses o hypo-
xia (O ega-Sáenz e al., 2010). In addi ion, se e al
g oups ha e shown ha modula ion o ol age-dependen
K+channels by hypoxia is main ained in glomus cells
dialysed wi h cons an (3–5 mM) le els o in acellula
ATP (López-López e al., 1989; Pé ez-Ga cía e al.,
2004).
Tocla i y hepo en ial oleo ATPinsignalling
acu e hypoxia, we ha e he ein analysed he sec e o y
esponseso glomuscells,inducedbyinhibi o so he
mi ochond ial ETC and oxida i e me abolism (such as
o enone and cyanide), in conjunc ion wi h CB s imuli
(hypoxia o glucose dep i a ion) which may comp omise
oxida i e phospho yla ion and ATP syn hesis. We ha e
also s udied he bioene ge ic cha ac e is ics o a glomus
cells and di ec ly moni o ed single-cell changes in cy o-
solic ATP le els in esponse o hypoxia and o he s imuli.
In pa allel s udies we ha e es ed whe he ecombinan
TASK3 channel ac i i y is modula ed by mi ochond ia
hypoxic signals (NADHandH2O2). Ou da a indica e ha
CB glomus cells ha e an ac i e oxida i e me abolism and
ha cy osolic ATP le els a e dec eased by ei he inhibi ion
o cy och ome c oxidase wi h cyanide o exposu e o
hecells o0mMglucose.Incon as ,acu ehypo-
xia does no induce dec eases in in acellula ATP. We
also demons a e ha ecombinan TASK3 channels a e
inhibi ed by in acellula NADH and H2O2.Theseda a
indica e ha cy osolic ATP does no pa icipa e in physio-
logical acu e O2sensing, al hough i may con ibu e o
hypoglycaemia signalling in glomus cells.
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
1094 M. To es-López and o he s J Physiol 603.5
Me hods
E hical app o al
All p ocedu es in his s udy we e app o ed by he
Ins i u ionalCommi ee o AnimalCa eandUsea he
Uni e si y o Se ille (PN2019 07/04/2020/051). Handling
o he animals was conduc ed in acco dance wi h he
Eu opean Communi y Council di ec i e o 22 Sep embe
2010 (Di ec i e 2010/63/EU) and he implemen a ions
o 5 June 2019 (Regula ion 2019/2010) o he ca e and
use o labo a o y animals. All in es iga o s unde s and
he e hical p inciples unde which he jou nal ope a es
and ha hei wo k complies wi h he animal e hics
checklis as ou lined in he jou nal policy. Fo in i o
expe imen s,animalswe ekilledbyi.p.injec iono ale hal
dose o anaes hesia (sodium hiopen al, 120–200 mg/kg)
o a oid su e ing be o e he animals we e killed and
issue dissec ion. Dea h was con i med by he comple e
cessa ion o mo emen and he absence o b ea hing o
o e 2 min. Ca o id bi u ca ions con aining he CB we e
emo ed immedia ely a e dea h. They we e hen ans-
e ed o a Pe i dish wi h cold (∼4°C) PBS placed in he
s ageo as e eomic oscope.A e ca e ulcleaningo he
su ounding issues, each CB was dissec ed and ex ac ed.
MCI-de icien mouse model
Mice (s ain B6/129) ca ying he loxed allele o
he Ndu s2 gene we e gene a ed in ou labo a o y
andb edwi h hoseca yingCRE ecombinaseunde
con ol o he y osine hyd oxylase (TH) p omo e o
gene a e emb yonic condi ional knockou mice wi h
dele ion o Ndu s2 in he ca echolamine gic issues
(Fe nández-Agüe a e al., 2015; Jiménez-Gómez e al.,
2023). Ndu s2 encodes a co e subuni o mi ochond ial
espi a o y chain NADH dehyd ogenase (MCI). We used
2–4-mon h-old MCI-de icien mice in his wo k.
Animal ca e
Expe imen al animals used in his s udy we e
4–8-week-oldWis a a s p oduced inou animal acili ies
and 2–4-mon h-old gene ically modi ied MCI-de icien
mice. Bo h emale and male animals in app oxima ely
hesamep opo ionwe eused.Animalswe ehouseda
22 ±1°C on a 12 h ligh /12 h da k cycle wi h ad libi um
access o ood (Teklad global 14% p o ein, En igo) and
wa e .
P epa a ion o CB slices
CB slices we e used o s udy he sec e o y ac i i y
o glomus cells by ampe ome y o o moni o he
cy osolic ATP/ADP a io and in acellula Ca2+by
mic o luo ime y. CB slices we e p epa ed as p e iously
desc ibed by ou g oup (Gao e al., 2021; Pa dal e al.,
2000; Pi ua e al., 2004). B ie ly, dissec ed CBs om a s
and mice we e embedded in 1% (w/ ) low-mel ing-poin
aga ose a 42°C p epa ed in PBS. Sec ions o he ca o id
bodies, 150 μm hick, we e cu using a ib a ome
(Leica, We zla , Ge many; VT1000S) and s o ed in
cold PBS. Mice CB slices we e diges ed o 5 min, a
37°C wi h shaking, in an enzyma ic solu ion con aining
50 μM CaCl2, 0.27 mg/ml ypsin (Sigma, S Louis,
MO, USA; ca . no. #T8003), 0.6 mg/ml collagenase
II (Sigma, ca . no. #C6885), and 1.25 U/ml po cine
elas ase (Millipo e, Bille ica, MA, USA; ca . no. #324682)
p epa ed in PBS pH 7.4. A e wa ds, eshly cu a
slices and diges ed mouse slices we e washed wice
wi hcoldPBSandincuba eda 37°Cin5%CO
2 o
24–48 h be o e use wi h he ollowing cul u e medium:
DMEM (0 glucose)/DMEM-F-12 (Gibco, Wal ham,
MA, USA; ca . no. #11966-025/21331-020) medium
(3:1) supplemen ed wi h 100 U/ml penicillin and
10 mg/ml s ep omycin (Gibco, ca . no. #15140-122),
2 mM l-glu amine (Gibco, ca . no. #25030-024), 10%
e al bo ine se um (FBS; Gibco, ca . no.#10270-10),
84 U/l insulin (Ac apid, No ono disk, Bags æ d,
Denma k; ca . no. #EU/1/02/230/011) and 1.2 U/ml
e y h opoie in (Sandoz, Basel, Swi ze land; ca . no. #EU
1/07/410/028).
Dissocia ion o CB cells
Dissocia ed a glomuscellswe eused ope o m
bioene ge ic s udies. Dissec ed CBs we e diges ed in he
same enzyma ic solu ion used o diges mouse CB slices
a 37°Cin5%CO
2 o 15 min wi hou shaking. Then,
he issue was mechanically s e ched wi h needles and
incuba ed o ano he 5 min. Diges ion was s opped by
headdi iono coldSeaho seassaymediumsupplemen ed
wi h 10% FBS o he enzyma ic solu ion. Seaho se
assay medium con ains Seaho se XF DMEM medium
supplemen ed wi h 3.5 mM glucose, 0.1 mM py u a e
and 4.8 mM l-glu amine. When s udying he e ec o
glucose dep i a ion on glomus cell ATP p oduc ion, no
glucose was added o he Seaho se assay medium. A e
cen i uga ion (5 min a 300 ga 4°C) he pelle was
washedwi h1mlo mediumandcen i ugedagain
o 5 min a 300 g. Finally, cells we e mechanically
dispe sed by pipe ing and he inal olume was adjus ed
o 70 μl. A e coun ing, cells we e seeded di ec ly in o
he Seaho se PDL mic opla e and cen i uged a 200 g o
2 min. Assay medium was added o each well o each a
inal olume o 180 μl. Cells we e incuba ed a 37°C o
45 min.
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
J Physiol 603.5 In acellula signalling in a e ial chemo ecep o s 1095
Bioene ge ic measu emen s in dispe sed glomus cells
Mi ochond ial and glycoly ic ATP p oduc ion a e in
a CB glomus cells was de e mined based on oxygen
consump ion a e (OCR) and ex acellula acidi ica ion
a e (ECAR) using Seaho se XFp Real-Time ATP a e
assay ki (Agilen Technologies, San a Cla a, CA, USA)
ollowing he manu ac u e ’s ins uc ion. Dissocia ed
glomus cells we e incuba ed in Seaho se assay medium
(see‘Dissocia iono CBcells’).Basalle elso OCRand
ECAR and hose in he p esence o mi ochond ial ETC
inhibi o s (1.5 μM oligomycin, 0.5 μM o enone and
an imycinA)we e henmeasu edinaSeaho seXFp
analyse (Seaho se Bioscience) in o de o calcula e ATP
p oduc ion a e. ATP p oduc ion a e was no malized by
he numbe o cells used in each measu emen .
Ampe ome ic eco ding o ca echolamine sec e ion
in single glomus cells
Measu emen o ca echolamines sec e ed by glomus cells
was pe o med as p e iously desc ibed in ou labo a o y
(O ega-Sáenze al.,2003).To eco d hesec e o yac i i y
o glomus cells, CB slices we e ans e ed o a eco ding
chambe con inuously pe used wi h ex acellula
solu ion (see ‘Reco ding solu ions’). Sec e o y e en s we e
eco ded wi h a 10 μm ca bon ib e pola ized a +750 mV
o de ec dopamine oxida ion. Ampe ome ic cu en s
we e eco ded wi h an EPC-7 pa ch-clamp ampli ie
(HEKA Elec onics, Lamb ech /P al z, Ge many), il e ed
a 100 Hz and digi ized a 250 Hz be o e s o age. Da a
acquisi ion was pe o med and analysed wi h an ITC-16
in e ace (Ins u ech Co po a ion, Bellmo e, NY, USA)
and PULSE/PULSEFIT so wa e (Heka Elec onics). The
sec e ion a e (pC/min) was calcula ed as he amoun o
cha ge ans e ed o he eco ding elec ode du ing he
las minu e o exposu e o he s imulus.
Two-pho on lase scanning mic oscopy imaging
Two-pho on lase scanning mic oscopy (2PLSM) was
used o measu e ATP/ADP a io and cy osolic Ca2+in
glomus cells in CB slices. Fluo escence was measu ed
using a Scien i ica mul ipho on gal anome e scanning
sys em (Scien i ica L d, Uk ield, UK) wi h an Olympus
Å∼60/1.0 NA wa e -dipping objec i e lens. A Chameleon
Ul a II (680–1080 nm), 3.5 W Ti:sapphi e lase sys em
(Cohe en lase g oup) p o ided he 2P exci a ion
sou ce. Lase powe a enua ion was achie ed using
wo Pockels cell elec o-op ic modula o s (S-MP-4700
and S-MP-6000-INT/UK) in se ies con olled by he
Lab iew so wa e (Scien i ica). Non-descanned emission
pho ons we e de ec ed wi h a GaAsP pho omul iplie ube
(PMT), S-MDU-PMT-50, g een, 490–560 nm. In ou
expe imen s wi h Pe ce alHR, he gene ically encoded
ATP/ADP senso , 2P exci a ion wa eleng hs o 820 nm
(ADP-sensi i e) and 950 nm (ATP-sensi i e) we e used.
Each wa eleng h equi ed sepa a e adjus men s be o e
s a ing he expe imen . Using he imaging so wa e,
powe and image acquisi ion se ings we e p elimina ily
adjus ed by s a ing wi h lowe se ings and g adually
inc easing lase powe o gain as necessa y. Imaging
beganin‘li e’mode oiden i y he egiono in e es
(ROI), whe e pa ame e s such as zoom, ield o iew,
esolu ion, dwell ime and ame a e we e op imized. Fo
expe imen s on CB slices, he imaging se ings we e se o
a esolu ion o 256 ×256 pixels, a zoom ac o o 4 and
10 μs dwell ime, es ic ing he ROI so ha he ame a e
wi h hesese ingsis3–4 .p.s.( amespe second).
Pe ce alHR was exp essed in CB cells using he
adeno-associa ed i al ec o (AAV9) con aining he
TH p omo e agmen embedded. Fo in ec ion, CB
slices we e incuba ed in comple e cul u e medium
supplemen ed wi h 2 ×1013 i al pa icles o 48 h
be o e he expe imen . Fo expe imen s, a CB slice was
ans e ed o he eco ding chambe and con inuously
pe used wi h ex e nal solu ion. To es ima e he
ATP/ADP a io using Pe ce alHR, he p obe was
exci ed wi h 950 and 820 nm ligh in apid succession
(González-Rod íguez e al., 2021; Tan ama e al., 2013).
G een channel (490–560 nm) luo escen emission
signals o bo h wa eleng hs we e de ec ed using a
non-descanned Scien i ica S-MDU-PMT-50-50 selec
GaAsP PMT. Two ime se ies o i e ames ( a e o
3–4 .p.s., 0.195 Å ∼0.195 mM pixels) we e acqui ed
o each wa eleng h. The a io alues om indi idual
dual acquisi ions we e eco ded as a ime lapse se ies.
A ime-se ies acquisi ion p o ocol was de eloped o
sequen ially swi ch he 2P lase wa eleng hs. Manual
adjus men s opowe andgainse ingswe emade o each
wa eleng h, wi h a 10 s s abiliza ion pe iod o he ligh
pa h be o e da a acquisi ion began. Lase powe and PMT
gain se ings we e adjus ed and sa ed o each wa eleng h
o ensu e baseline luo escence was b igh bu below he
sa u a ion h eshold o 4095 luo escence uni s. Baseline
luo escence was adjus ed o ∼800 uni s, wi h back-
g ound luo escence ypically anging om 100 o 150
uni s. To ensu e accu a e measu emen o luo escence
changes, he baseline a io be ween luo escence a 950
and 820 nm was main ained nea 1. This acili a ed
moni o ing du ing he expe imen and simpli ied sub-
sequen da a analysis. Dual-wa eleng h acquisi ions we e
collec ed e e y 3 min du ing ime-se ies expe imen s,
wi h i e ames pe wa eleng h acqui ed and a e aged o
analysis. The expe imen s aimed o examine he e ec s
o a ious ea men s o s imuli on he cellula ATP/ADP
a io. Du ing me abolic manipula ions, luo escence
le els a 950 and 820 nm ypically exhibi ed opposi e
di ec ional changes. Time se ies analysis was conduc ed
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
1096 M. To es-López and o he s J Physiol 603.5
o line using FIJI. Mul iple cy osolic ROIs and a back-
g ound ROI we e measu ed and sub ac ed. The 950/820
a io was calcula ed o each ROI a each ime poin .
Thecon ibu iono mi ochond ia o hebioene ge ic
s a us o each cell ( he OXPHOS index) was es ima ed
by compa ing he dec ease in he Pe ce alHR ATP/ADP
a ioinducedbyba happlica iono oligomycin(10μM)
wi h he one induced by addi ion o oligomycin (10 μM)
wi h 2-deoxy-glucose (2-DG), a non-hyd olysable sub-
s i u e o glucose.
Fo cy osolic Ca2+measu emen CB slices we e
loaded wi h 10 μM Fluo-3/AM (In i ogen, Ca lsbad,
CA, USA; F1242) in Ty ode solu ion wi hou calcium
a 37°C o 15 min, ollowed by 10 min a oom
empe a u e. A e loading, CB slices we e ans e ed o
he eco ding chambe and con inuously pe used wi h
ex e nal solu ion. The p obe was exci ed a 950 nm o
analyse he Ca2+signal using Fluo-3/AM. G een channel
(490–560 nm) luo escen emission signals we e de ec ed
using a non-descanned Scien i ica S-MDU-PMT-50-50
selec GaAsPPMT. Cy osolic[Ca2+]signalswe edigi ized
a a sampling in e al o 500 ms.
Immunohis ochemis y
CB slices exp essing TH-Pe ce alHR a e 48 h o
in ec ion we e ixed wi h 4% pa a o maldehyde in PBS
o 30 min a oom empe a u e, pe meabilized wi h
PBS-T i on X (PBS-T) 0.1%, blocked wi h blocking se um
solu ion o 1ha oom empe a u eandincuba ed
wi h he TH (No us Biological, Li le on, CO, USA;
ca . no. #BD300-109, RRID: AB_10077691, 1:1000) and
g een luo escen p o ein (GFP) (A esLab, Da is, CA,
USA; ca . no. #GFP-1202, RRID:AB_2734732, 1:400)
p ima y an ibodies o e nigh a 4°C. Finally, slices we e
incuba ed o 2 h a oom empe a u e wi h he Alexa568
(In i ogen, ca . no. #A10042, RRID:AB_2534017,
1:400) and Alexa488 (In i ogen, ca . no. #A11039,
RRID:AB_2534096, 1:400) seconda y an ibodies and
nuclei we e s ained wi h 4,6-diamidino-2-phenylindole
(DAPI). Immuno luo escen images we e acqui ed
using a Leica S ella is 8 con ocal mic oscope (Leica,
RRID:SCR_02 4664).
Pa ch clamp eco dings om HEK cells exp essing
TASK3 channels
HEK293 cells we e main ained in high glucose and
py u a e DMEM medium (Gibco, ca . no. #21969-035)
supplemen ed wi h 10% FBS (Gibco, ca . no. #10270-10),
1% penicillin/s ep omycin (Gibco, ca . no. #15140-122)
and 1% l-glu amine (Gibco, ca . no. #25030-024). Cells
we e g own as a monolaye in he bo om o 100 mm
Pe i dishes o 70% con luence on he day o ans-
ec ion and washed wi h PBS. Cells we e dissocia ed by
adding 2 ml ypsin o he Pe i dish and incuba ing o
2 min a 37°C. A e wa ds, 9 ml o supplemen ed DMEM
was used o esuspend cells p e ious o cen i uga ion
a 200 g o 5 min in a 15 ml ube. The pelle was
esuspended in 1 ml DMEM and cells we e coun ed
using ypan blue solu ion. In o al, 15 ×104cells o
each ans ec ion we e pelle ed again, esuspended in
15 μl o Neon Resuspension Bu e R (Neon ans ec ion
sys em, In i ogen, ca . no. #MPK10096) and mixed
wi h 2 μg plasmid DNA (pcDNA3.1) encoding TASK3
o GFP. Elec opo a ion was ca ied ou acco ding o
he manu ac u e ’s ins uc ions. Resuspended cells we e
pulsed wicewi ha ol ageo 1.100Vandawid ho 20ms.
A e elec opo a ion cells we e pla ed on glass co e slips
ea edwi hpoly-l-lysineandkep in hesamecul u e
medium a 37°C in a 5% CO2and 21% O2incuba o o
24 h.
Mac oscopic ionic cu en s we e eco ded a oom
empe a u e om HEK cells ansien ly ans ec ed wi h
TASK3 and/o GFP using he whole cell con igu a ion
o he pa ch clamp echnique. Pa ch clamp pipe es
had esis ances anging om 2 o 4 Mwhen illed
wi h he in e nal solu ion (see ‘Reco ding solu ions’).
Vol age-clamp eco dings we e ob ained wi h an EPC-10
ampli ie wi h an in eg a ed AD in e ace (ITC-1600,
HEKA). Da a acquisi ion and analysis was pe o med
using he Pa chMas e /Fi Mas e so wa e (HEKA). HEK
cells we e held a −70 mV and hen subjec ed o
di e en pulse p o ocols gene a ed by he Pa chMas e
so wa e.
Reco ding solu ions
Fo ampe ome ic and mic o luo ime ic eco dings,
slices we e ans e ed o a eco ding chambe
con inuously pe used wi h con ol bica bona e bu e ed
solu ion, con aining (in mM) 125 NaCl, 23 NaHCO3,
4.5 KCl, 1 MgCl2,2.5CaCl
2, 5 suc ose and 5 glucose. In
he high K+solu ion, KCl subs i u es NaCl equimola ly.
Fo he low glucose expe imen s, cells we e exposed
o a glucose- ee solu ion, whe e suc ose eplaced
glucose (in mM, 125 NaCl, 23 NaHCO3, 4.5 KCl, 1
MgCl2,2.5CaCl
2and 10 suc ose). The 2-DG solu ion
was glucose/suc ose- ee and con ained 5 mM 2-DG.
Pha macological agen s we e added o he con ol
solu ion a he desi ed concen a ion when equi ed.
The ‘no moxic’ solu ions we e con inuously bubbled
wi h a gas mix u e con aining 5% CO2, 20% O2and 75%
N2(O2 ension ∼145 mmHg). The ‘hypoxic’ solu ions
we e bubbled wi h a gas mix u e o 5% CO2and 95% N2,
eaching an O2 ension o ∼10–15 mmHg in he eco ding
chambe . Osmolali y o solu ions was ∼300 mOsm/kg
wi h pH 7.4. Expe imen s we e pe o med a ∼35°C.
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
J Physiol 603.5 In acellula signalling in a e ial chemo ecep o s 1097
Comple e eplacemen o solu ions in he chambe was
achie ed in ∼60 s.
Fo pa ch-clamp eco dings, pipe es we e illed wi h
in e nal solu ion con aining (in mM) 150 KCl, 3 MgCl2,
10 HEPES and 5 EGTA (pH 7.2 adjus ed wi h KOH). The
composi ion o he ba h solu ion was (in mM) 145 NaCl,
2.5 KCl, 3 MgCl2,1CaCl
2,10glucoseand10HEPES(pH
was adjus ed using HCl o NaOH o he desi ed alues).
Tos udy hee ec o NADHandH
2O2on mac oscopic
cu en s, hese agen s we e added o he in e nal solu ion
a he desi ed concen a ion (NADH: 200 and 400 μM,
H2O2: 200 and 500 μM).
S a is ical analysis
S a is ical analysis was pe o med using P ism
Ve sion 8.2.1. (279) o MacOS. No mali y o he
da a ob ained in he expe imen s was assessed wi h
Shapi o-Wilk, D’Agos ino & Pea son, Ande son-Da ling
and Kolmogo o -Smi no es s. In some cases, a log
ans o ma ion o he da a was ca ied ou o no malize
he dis ibu ion p io o pa ame ic analysis. Fo g aphical
ep esen a ion o he da a wi h a no mal dis ibu ion, we
used ba diag ams wi h a sca e plo o da a poin s supe -
imposed. These da a a e gi en as mean ±SD and he
numbe (n) o expe imen s. Fo g aphical ep esen a ion
o non-pa ame ic da a, we used box plo s whe e median,
qua iles and whiske s (highes and lowes alues)
a e indica ed. Compa isons be ween wo g oups we e
pe o med using pai ed o unpai ed es s o da a wi h a
no mal dis ibu ion, and Wilcoxon o a Mann–Whi ney
es o non-pa ame ic da a. Compa isons o mul iple
g oups wi h a no mal dis ibu ion we e done using
a one-way ANOVA ollowed by Tukey’s mul iple
compa isons pos hoc es s. Mul iple non-pa ame ic
da a we e compa ed using K uskal–Wallis ollowed by
Dunn’s mul iple compa ison es s. S a is ical es s used
a e also indica ed in he igu e legends. P<0.05 was
conside ed s a is ically signi ican and he esul ing alue
is ep esen ed in each igu e panel.
Resul s
Blocking MCI abolishes esponsi eness o glomus
cells o hypoxia bu no o cyanide
Hypoxiainglomuscellsisknown op oduce
dose-dependen inc eases in he mi ochond ial
p oduc ion o NADH and ROS and a neu osec e o y
esponse, which a e mimicked by blocking MCI wi h
o enone. Ro enone and he gene ic dis up ion o
MCI a e bo h able o abolish any e ec o hypoxia
on mi ochond ial signalling o cell sec e o y ac i i y,
indica ing ha a unc ional MCI is essen ial o acu e
O2sensing by glomus cells (Fe nández-Agüe a e al.,
2015; O ega-Sáenz e al., 2003). Suppo ing his model
(Fig. 1A), in ampe ome ic expe imen s pe o med
he e, hypoxia and o enone sepa a ely elici ed clea
inc eases in he sec e o y ac i i y o glomus cells in CB
slices. Howe e , esponsi eness o hypoxia was no only
abolished by o enone bu o enone-induced sec e o y
ac i i y also dec eased du ing hypoxia (Fig. 1B,C,F). This
inding, which is eminiscen o he hypoxic inhibi ion
o o enone-induced ROS p oduc ion in mi ochond ia
(A ias-Mayenco e al., 2018), con as s wi h wha would
be expec ed i a dec ease in in acellula ATP (indica ed
by he ques ion ma k in Fig. 1A)pa icipa esinbo h
o enone and hypoxia signalling.
I couldbea gued ha he lowo elec ons ed ocy o-
ch ome c oxidase is d as ically educed in he p esence
o o enone and ha unde hese condi ions hypoxia has
no e ec because i canno p oduce u he inhibi ion o
MCIV. To es his hypo hesis, we eco ded he sec e o y
ac i i y induced by cyanide in cells al eady exposed o
o enone o se e al minu es. Cyanide is a po en MCIV
blocke which has no e ec on glomus cells when he ETC
is in e up ed (e.g. due o gene ic dis up ion o MCIII)
and elec ons do no each MCIV (Cabello-Ri e a e al.,
2022). In all cells es ed (n=11), applica ion o cyanide in
he p esence o o enone induced a po en and e e sible
sec e o y esponse (Fig. 1D–F). These da a indica e ha
MCIV is s ill unc ional and inhibi able by cyanide in cells
ea ed wi h o enone o se e al minu es. Howe e , e en
wi ha unc ionalMCIV,blockingMCImakesglomus cells
un esponsi e o hypoxia.
Di e en ial e ec o MCI dis up ion on sensi i i y o
hypoxia and hypoglycaemia
TheCBisoneo heo ganswi h hehighes O
2
consump ion o which glucose p o ides he uel
o oxida i e phospho yla ion. On he o he hand,
glomuscellsa eac i a edbyhypoxiaandhypoglycaemia
(Ga cía-Fe nández e al., 2007; Pa dal & López-Ba neo,
2002; Zhang e al., 2007). We compa ed he e ec o
inhibi ion o he ETC wi h o enone on he esponsi eness
o a glomus cells o hypoxia and glucose dep i a ion.
Whe eas he sec e o y esponse o cells o hypoxia was
occluded in he p esence o o enone, esponsi eness
o 0 mM glucose was main ained (Fig. 2A,B). Simila
di e ences in esponses o hypoxia and low glucose
in he p esence o o enone we e obse ed in glomus
cells o mice (Fig. 2C,D). Mo eo e , glomus cells om
gene ically modi ied mice lacking a unc ional MCI in he
ca echolamine gic e i o y (see Fe nández-Agüe a e al.,
2015; Jiménez-Gómez e al., 2023) we e un esponsi e
o hypoxia bu showed a obus sec e o y esponse o
glucose dep i a ion, which was e en highe han in
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
1098 M. To es-López and o he s J Physiol 603.5
wild- ype glomus cells (Fig. 2E–H). In MCI-de icien
glomus cells ob ained om gene ically modi ied mice,
sensi i i y o glucose dep i a ion is o ally independen
o oxida i e phospho yla ion because in hese cells,
in which MCI has been dis up ed o o e 2 mon hs,
mi ochond ial ETC and oxida i e ATP syn hesis a e
s ongly inhibi ed as hey a e insensi i e o cyanide,
al hough main ain no mal esponses o CO2(Fig. 2I–L)
(see also Jiménez-Gómez e al., 2023). The ac ha
sensi i i y o hypoxia and glucose dep i a ion (bo h
comp omising oxida i e phospho yla ion) depends on
sepa a e pa hways di e en ially a ec ed by o enone
o gene ic MCI dis up ion u he sugges s ha
changes in mi ochond ial p oduc ion o ATP a e
no in ol ed in acu e hypoxia signalling. Howe e ,
modi ica ions in non-oxida i e cy osolic ATP le els could
con ibu e o he esponsi eness o glomus cells o low
glucose.
0.0009
D
A
MCI
MCII
CoQH2MCIII MCIV
O2
H2O
Hypoxia
NADH
ROS ROS
Ro CN
ATP?
Hypoxia
Ro enone
Hypoxia CN
5 pA
B
5 pA
1 min
Hypoxia
Ro enone
Hypoxia
1 min
1 min
2 pA
Ro enone
CN CN
0
5
10
15
Sec e ion a e (pC/min)
C
Ro Ro
Hx
E
F
0.0106
0
5
10
15
Sec e ion a e (pC/min)
20
25
Ro Ro
CN
Figu e 1. Blockade o MCI abolishes esponsi eness o glomus cells o hypoxia bu no o cyanide.
A, scheme illus a ing he elec on anspo chain (ETC) and i s dynamic changes in esponse o hypoxia ( ed
lines and symbols). No e he accumula ion o educed coenzyme Q (CoQH2), NADH and ROS du ing exposu e
o hypoxia. The inhibi ion o mi ochond ial complex I (MCI) and IV (MCIV) by o enone (Ro ) and cyanide (CN)
a e illus a ed in g ey and blue, espec i ely. B, ep esen a i e ampe ome ic eco ding o ca echolamine sec e ion
o glomus cells in a CB slices in esponse o hypoxia (O2 ension ∼15 mmHg), o enone (1 μM) and o enone
+hypoxia. C, a e age sec e ion a e induced by o enone (Ro , 1–5 μM) and o enone +hypoxia (Ro +Hx) in
a CB glomus cells. Values in pC/min a e: Ro (5.4 ±3.1, n = 8 cells/5 a s), Ro +Hx (1.6 ±0.8, n = 8 cells/5
a s). D, ep esen a i e ampe ome ic aces eco ded om a CB glomus cells in slices in esponse o cyanide
(CN, 300 μM), o enone (1 μM) and o enone +cyanide. E, a e age sec e ion a e induced by o enone (Ro
1–5 μM) and o enone +cyanide (Ro +CN) in a CB glomus cells. Values in pC/min a e: Ro (3.5 ±3.5, n = 7
cells/3 a s), Ro +CN (11.6 ±5.7, n = 7 cells/3 a s). F, ep esen a i e ampe ome ic eco ding (ou o ou simila
expe imen s) o ca echolamine sec e ion o glomus cells om a a CB slice in esponse o hypoxia, o enone (1 μM),
o enone +hypoxia and o enone +cyanide (300 μM). No e he dec ease in he sec e o y ac i i y when he cell
is exposed simul aneously o o enone and hypoxia. By con as , inhibi ion o MCIV by cyanide in he p esence o
o enone s ongly ac i a es he cell. Da a a e exp essed as mean ±SD wi h all da a alues supe imposed. P- alues
(<0.05) calcula ed by a wo- ailed pai ed es a e indica ed in he panel.
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
J Physiol 603.5 In acellula signalling in a e ial chemo ecep o s 1099
0.0282
Basal
Sec e ion a e (pC/min)
20
15
10
5
0Ro
< 0.0001
Ro
0 Glu
<0.0001
Sec e ion a e (pC/min)
CN
Sec e ion a e (pC/min)
Basal CN
10
5
0
Basal
Con ol
EHypoxia 0 Glu
5 pA
40K
1 min
ARo enone
5 pA
1 min
Hypoxia 0 Glu
Hypoxia 0 Glu
B
Sec e ion a e (pC/min)
10
5
00 Glu
F
MCI-de icien
Hypoxia
0 Glu
5 pA
40K
1 min
G
Sec e ion a e (pC/min)
20
10
0
0 Glu
Basal
<0.0001
H
Ra
Mouse
Con ol MCI-de icien
Hypoxia CN
5 pA
1 min
IJK
CNHypoxia
5 pA
CO2CO2
1 min
L
C
D
Sec e ion a e (pC/min)
15
10
5
0Ro Ro
0 Glu
1 min
Ro enone
5 pA
Hypoxia
0 Glu
Hypoxia
0 Glu
hypoxia
0 Glu
0.0008
15
10
5
0
15
Hx Hx
0.0035
0.537
0.121
>0.999
>0.999
Figu e 2. Di e en ial e ec o MCI dis up ion on sensi i i y o hypoxia and glucose dep i a ion.
A, ep esen a i e ampe ome ic aces o he sec e o y ac i i y eco ded om a CB glomus cells in slices in
esponse o hypoxia (O2 ension ∼15 mmHg), 0 mM glucose (0 Glu), o enone (1 μM), o enone +hypoxia and
o enone +0 Glu. B, a e age sec e ion a e (pC/min) induced by o enone (Ro , 1–5 μM) and o enone +0glucose
(Ro +0 Glu) in a CB glomus cells. Da a a e exp essed as mean ±SD wi h all da a alues supe imposed. Values
in pC/min a e: Ro (4.7 ±3.2, n = 19 cells/11 a s) and Ro +0 Glu (12.1 ±3.7, n = 6 cells/3 a s). P- alue (<0.05)
calcula ed by unpai ed wo- ailed es is indica ed. C, ep esen a i e ampe ome ic aces o sec e o y ac i i y
eco ded om mouse CB glomus cells in slices in esponse o hypoxia (O2, ension∼15 mmHg), 0 mM glucose (0
Glu), o enone (5 μM), o enone +hypoxia and o enone +0 Glu. D, a e age sec e ion a e (pC/min) eco ded om
con ol mouse glomus cells in esponse o o enone (Ro , 5 μM) and o enone +0 glucose (Ro +0 Glu). Values in
pC/min a e: Ro (2.3 ±2.3, n = 7 cells/4 mice), Ro +0 Glu (10.1 ±3.5, n = 7 cells/4 mice). Compa ison be ween
bo h g oups was pe o med using a pai ed wo- ailed es . P- alue is indica ed in he panel. E and G, ep esen a i e
eco dings o he sec e o y ac i i y induced by hypoxia, glucose dep i a ion (0 Glu) and high K+in glomus cells
in CB slices om con ol mice (E) and MCI-de icien mice (G). F and H, quan i ica ion o he sec e o y ac i i y
(pC/min) eco ded in basal condi ions and du ing 0 Glu in glomus cells om con ol mice (F; n = 18 cells/15 mice)
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
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Addi ional in o ma ion
Da a a ailabili y s a emen
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The au ho s ha e no con lic s o in e es o decla e.
Au ho con ibu ions
M.T.-L., P.G.-R., O.C., H.-S. R., H.T.-T., A.C., L.G., P. O.-S. and
J. L.-B. pe o med he expe imen s and pa icipa ed in he in e -
p e a ion o da a. M-T.-L., P.G.-R., O.C., L.G., P. O.-S. and J.L.-B.
designed he igu esandw o e he i s d a o hepape .P.
O.-S. and J.L.-B w o e he inal d a o he pape and supe ised
he p ojec . All au ho s ead and app o ed he inal pape . All
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ag ee obeaccoun able o allaspec so hewo k.Allpe sons
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quali y o au ho ship a e lis ed.
Funding
This esea ch was suppo ed by Spanish Minis ies o Science
and Inno a ion and Heal h (G an s PID2019-106410RB-
I00, PID2019-110817R, PID2022-138131OB-I00 and PID2023-
146862OB-100 unded by MCIN/AEI/10.13039/501100011033
o J.L.-B., L.G. and P.O.-S) and he Eu opean Resea ch Council
(ERC Ad anced G an PRJ201502629).
Acknowledgemen s
We hank Al edo Ca o-Maldonado (Agilen Technologies) o
his echnical help in se ing up he me abolic analysis using he
Seaho se analyse . We also hank echnical help p o ided by he
s a a he Co e Facili ies o Ins i u o de Biomedicina de Se illa
(IBiS).
Keywo ds
acu e oxygen sensing, ca o id body glomus cells, cy osolic
ATP, elec on anspo chain inhibi o s, glucose sensing,
H2O2,hypoxia,mi ochond ia- o-memb anesignalling,NADH,
TASK3 channels
Suppo ing in o ma ion
Addi ional suppo ing in o ma ion can be ound online in he
Suppo ing In o ma ion sec ion a he end o he HTML iew o
he a icle. Suppo ing in o ma ion iles a ailable:
Pee Re iew His o y
© 2025 The Au ho (s). The Jou nal o Physiology published by John Wiley & Sons L d on behal o The Physiological Socie y.
14697793, 2025, 5, Downloaded om h ps://physoc.onlinelib a y.wiley.com/doi/10.1113/JP287130 by Readcube (Lab i a Inc.), Wiley Online Lib a y on [16/06/2025]. See he Te ms and Condi ions (h ps://onlinelib a y.wiley.com/ e ms-and-condi ions) on Wiley Online Lib a y o ules o use; OA a icles a e go e ned by he applicable C ea i e Commons License
