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Tiêu đề Endovanilloids putative endogenous ligands of transient receptor potential vanilloid 1 channels
Tác giả Mario Van Der Stelt, Vincenzo Di Marzo
Trường học Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche
Chuyên ngành Biochemistry
Thể loại Minireview
Năm xuất bản 2004
Thành phố Pozzuoli
Định dạng
Số trang 8
Dung lượng 240,17 KB

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M I N I R E V I E WEndovanilloids Putative endogenous ligands of transient receptor potential vanilloid 1 channels Mario van der Stelt and Vincenzo Di Marzo Endocannabinoid Research Grou

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M I N I R E V I E W

Endovanilloids

Putative endogenous ligands of transient receptor potential vanilloid 1 channels

Mario van der Stelt and Vincenzo Di Marzo

Endocannabinoid Research Group, Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, Pozzuoli, Italy

Endovanilloids are defined as endogenous ligands of the

transient receptor potential vanilloid type 1 (TRPV1)

protein,a nonselective cation channel that belongs to the

large family of TRP ion channels,and is activated by

the pungent ingredient of hot chilli peppers,capsaicin

TRPV1 is expressed in some nociceptor efferent neurons,

where it acts as a molecular sensor of noxious heat and

low pH However,the presence of these channels in

var-ious regions of the central nervous system,where they are

not likely to be targeted by these noxious stimuli,suggests

the existence of endovanilloids Three different classes of

endogenous lipids have been found recently that can

activate TRPV1,i.e unsaturated

N-acyldopamines,lip-oxygenase products of arachidonic acid and the

endo-cannabinoid anandamide with some of its congeners To classify a molecule as an endovanilloid,the compound should be formed or released in an activity-dependent manner in sufficient amounts to evoke a TRPV1-mediated response by direct activation of the channel To control TRPV1 signaling,endovanilloids should be inactivated within a short time-span In this review,we will discuss, for each of the proposed endogenous ligands of TRPV1, their ability to act as endovanilloids in light of the criteria mentioned above

Keywords: anandamide; arachidonic acid; cannabinoid; lipid; signaling; TRP; VR1

Introduction

The transient receptor potential vanilloid type 1 (TRPV1)

protein is a nonselective cation channel that belongs to the

large family of TRP ion channels and is highly expressed in

small diameter primary afferent fibers [1] It is a molecular

integrator of noxious stimuli,such as heat and low pH,

and can also be activated by the pungent ingredient of hot

chilli peppers,capsaicin,as well as by other plant toxins,the

most potent of which is resiniferatoxin (RTX) [2] In

primary sensory neurons,TRPV1 is essential for the

development of inflammatory hyperalgesia [3,4] Although

initially controversial,it has now been firmly established

that TRPV1 is synthesized in cells outside the peripheral nervous system,such as keratinocytes,epithial and endo-thelial cells,where it serves as yet undefined purposes [5] Furthermore,TRPV1 has also been found in various brain areas,including dopaminergic neurons of the substantia nigra,hippocampal pyramidal neurons,hypothalamic neu-rons,the locus coeruleus in the brainstem and in various layers of the cortex,where it might be involved in modulation of synaptic plasticity [6,7]

In the central nervous system or under normal physio-logical conditions,TRPV1 is unlikely to be activated by heat or low pH,therefore it has been suggested that other endogenous ligands of this ion channel exist Indeed,three different classes of lipids,all derived from the metabolism

of arachidonic acid (AA),have been recently characterized that can activate TRPV1,i.e in chronological order,the endocannabinoid anandamide,some lipoxygenase products

of AA,and N-arachidonoyldopamine (Fig 1) To qualify as

an endovanilloid,i.e an endogenous activator of TRPV1, the compound should be formed by cells and be released in

an activity-dependent manner in sufficient amounts to evoke

a TRPV1-mediated response by direct binding and subse-quent activation of the channel Finally,endovanilloid signalling should be terminated within a short time-span to allow a strict regulation of its actions Therefore,biosyn-thetic and metabolic pathways for a putative endovanilloid should be present in close proximity to TRPV1 As the putative binding site for an endogenous ligand at TRPV1

is intracellular [8,9], it might be expected that putative endogenous ligands are produced inside the cell or,in the case of production at a more distant site,brought into the cell In this review we will discuss the capabilities of

Correspondence to V Di Marzo,Instituto di Chimica Biomolecolare,

Consiglio Nazionale delle Ricerche,Via Campi Flegrei 34,

Comprensorio Olivetti,Bldg 70 80078 Pozzuoli (NA),Italy.

Fax: + 39 081 8041770,Tel.: + 39 081 8675093,

E-mail: vdimarzo@icmib.na.cnr.it

Abbreviations: 12-HETE,12-hydroxyeicosatetraenoic acid; 12S- and

15S-HPETE,12-(S)- and 15-(S)-hydroperoxyeicosatetraenoic acid;

AA,arachidonic acid; DRG,dorsal root ganglia;

RTX,resinifera-toxin; TRPV1,transient receptor potential vanilloid type 1 protein.

Note added during revision: During the revision process of this article,

NADA was reported to exert a potent vasodilation of rat mesenteric

arteries via mechanisms including TRPV1 receptors as well as

cann-abinoid CB1 receptors and an as-yet-uncharacterized canncann-abinoid

endothelial receptor [O’Sullivan,S.E.,Kendall,D.A & Randall,M.D.

(2004) Br J Pharmacol., 141,803–812].

(Received 16 January 2004,revised 13 February 2004,

accepted 18 February 2004)

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N-arachidonoyldopamine,lipoxygenase products of AA

and anandamide to act as endogenous activators for TRPV1

in vivoin light of the above mentioned criteria

N-arachidonoyldopamine

Biosynthesis

N-arachidonoyldopamine was characterized originally in

the striatum of bovine brain and its distribution in the rat

nervous system is as follows (greatest abundance to lowest):

striatum > hippocampus > cerebellum > thalamus >>

dorsal root ganglia (DRG) [10] The basal levels of

N-arachidonoyldopamine are low (6 pmolÆg)1 striatum)

and its stimulus-evoked formation has yet to be

demon-strated [10] Noteworthy,other congeners of this compound

have been detected recently in the bovine brain,i.e

N-oleoyldopamine, N-palmitoyldopamine and

N-stearoyl-dopamine [11] Of these, N-oleoylN-stearoyl-dopamine also possesses

the ability to activate TRPV1 with the same potency as

N-arachidonoyldopamine [11] N-arachidonoyldopamine

can theoretically be formed through two biosynthetic

pathways [10] Firstly,a direct condensation of AA (or its

Coenzyme A derivative) with dopamine has been proposed,

but the enzyme responsible for this condensation has not

yet been characterized The second route is an indirect

pathway in which N-arachidonoyltyrosine is converted

into N-arachidonoyldopamine Interestingly, TRPV1 mRNA was colocalized with mRNA of tyrosine hydroxy-lase in dopaminergic neurons of the substantia nigra, which project to the striatum [6] To date,conclusive experimental evidence for either pathway is still lacking

If N-arachidonoyldopamine acts in a paracrine manner,

it has to be transported into the cell to reach its site of action In support to this hypothesis, N-arachidonoyl-dopamine was rapidly taken-up by C6 glioma cells via the rapid facilitated diffusion process [10] that is also respon-sible for anandamide transport across membranes (reviewed

in [12])

Degradation The same membrane transport process carrying N-arachi-donoyldopamine into the cell can theoretically also be used

to extrude it from the cell,thereby terminating its actions

at TRPV1 Other possible inactivation pathways are the hydrolysis of its amide bond by a hydrolase or its methylation of the hydroxyl group of its cathechol moiety

by a cathechol-O-methyl-transferase (Fig 2) The latter route might represent the inactivation pathway in in vivo nervous tissues where this enzyme is abundant because N-arachidonoyldopamine was only very slowly hydrolysed

by brain homogenates and its methylated derivative was less potent or inactive at TRPV1 [10]

Fig 1 Chemical structures of endogenous TRPV1 ligands.

Fig 2 Metabolic pathways of N-arachido-noyldopamine.

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Pharmacology and physiological actions

N-arachidonoyldopamine is a full agonist of TRPV1 and

the most potent endogenous lipid ligand discovered to date

It has a Kd of approximately 5–10 lM in binding assays

performed in heterologous or native expression systems,

respectively [13,14] It is five to 10-fold more potent than

anandamide and almost equipotent as capsaicin in some

functional assays,with an EC50 in Ca2+-influx assays of

approximately 50 nM at human and rat TRPV1

over-expressed in HEK293 cells N-arachidonoyldopamine is

also able to rapidly desensitize the TRPV1 receptor to

subsequent capsaicin activation [10] Its potency seems to be

dependent on the cell and assay conditions used to assess its

functional activity,because in rat DRG neurons (using

Ca2+imaging) and rat TRPV1-CHO cells (using a Ca2+

-uptake assay) its potency is in the low micromolar range

(0.8–5 lM) [10,13] Exogenous N-arachidonoyldopamine

induces,in a TRPV1-dependent manner,calcitonin

gene-related peptide and substance P release from spinal cord

slices and enhances hippocampal paired-pulse depression

(EC50 0.1–0.2 lM [10]) It is also able to induce

TRPV1-mediated thermal hyperalgesia [10],and constriction of

isolated bronchi and urinary bladder [15],albeit,in these

two assays N-arachidonoyldopamine is much less potent

and efficacious than capsaicin Species differences and

pharmacodynamic factors (such as reduced bioavailability

and limited access to the intracellular site on TRPV1) were

suggested to be responsible for these varying potencies [15]

As yet,no TRPV1-mediated physiological or pathological

conditions have been attributed to endogenously formed

N-arachidonoyldopamine

Lipoxygenase products of arachidonic acid

Biosynthesis

Various oxygenated AA derivatives were shown to activate

TRPV1 [16] 12-(S)- and

15-(S)-hydroperoxyeicosatetrae-noic acid (12S- and 15S-HPETE) possessed the highest

potency These AA metabolites can be produced by

lipoxygenases that introduce molecular oxygen in a

regio-and stereoselective dependent manner in AA released from

the membrane by phospholipase A2enzymes (Fig 3) [17]

Several isoforms for both 12S- and 15S-lipoxygenases have

been identified 15(S)-Lipoxygenase-1 has been found in

human eosinophils and a second

isoform,15(S)-lipoxyge-nase-2,is expressed in the prostate,lung,cornea and hair

roots [18] There are three 12-lipoxygenase isoforms that

have a broad tissue distribution,including the lung,pineal

gland,spleen,macrophages and keratinocytes [19]

12S-Lipoxygenase has also been found in the canine brain,

including basal ganglia,hippocampus and hypothalamus

[20,21] Importantly, 12-lipoxygenase mRNA was found

to be coexpressed with TRPV1 in rat DRG neurons [22]

However,colocalization with TRPV1 in neurons of the

central nervous system has not been studied yet

Lipoxygenases are not stimulated by calcium,but their

substrate AA can be liberated from the membrane by an

agonist-stimulated increase in intracellular calcium that

activates Ca2+-dependent phospholipases A2 [17] For

example,it was shown that the potent inflammatory

mediator bradykinin could induce the production of 12-hydroxyeicosatetraenoic acid (12-HETE),a metabolic product of 12-HPETE,from rat primary sensory neurons [22] Furthermore,AA can be produced from anandamide

by hydrolysis of its amide bond by fatty acid amide hydrolase [23] Interestingly,12/15-lipoxygenases can also accept AA esterified into membrane lipids and the endo-cannabinoid anandamide as a substrate [18,24,25–27] The lipoxygenase products of anandamide are potent endo-genous inhibitors of fatty acid amide hydrolase [27,28] and might stimulate TRPV1 themselves [29,30], although attempts to demonstrate their direct activity at these receptors have given negative results [8,31]

Degradation The hydroperoxy group in the lipoxygenase products of AA

is very labile and easily reduced into a hydroxy-group by glutathione peroxidases,which might constitute an inacti-vation pathway,because the reduced lipoxygenase products are far less potent on TRPV1 (Fig 3) [16]

Pharmacology and physiological actions 12-HPETE could displace [3H]RTX from recombinant TRPV1-HEK293 cells more potently than capsaicin (Ki¼ 0.35 and 2.5 lM,respectively) [22] However,in inside-out patches of neonatal rat dorsal root ganglia neurons 12(S)- and 15(S)HPETE had an EC50 values of approximately 8 lM,which was seven- to eightfold higher than that for capsaicin as measured by single channel current recordings [16] To date,no pharmacological study has described the activity of exogenous lipoxygenase products on typical TRPV1-mediated responses in vivo or

Fig 3 Biosynthetic and metabolic pathways of 12- and 15-HPETE Pathways indicated (a) under physiological conditions and during pharmacological assays and (b) during pharmacological assays.

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ex vivo,such as hyperalgesia,and vasodilation or

broncho-or urinary bladder constriction This lack of data might

be due to practical problems such as the instability of

these compounds However,some studies indirectly suggest

that endogenously produced lipoxygenase metabolites

are responsible for TRPV1-mediated actions in vivo For

example,protease-activated receptor-2 activation causes

endothelium-dependent coronary vasodilation in a

capsaze-pine-sensitive manner,which might be mediated through a

lipoxygenase metabolite,because three structurally different

lipoxygenase inhibitors were able to attenuate this response

[32] In their elegant study,Shin et al provided evidence

that 12(S)-HPETE is produced endogenously in sensory

neurons upon stimulation of sensory nerve endings by the

inflammatory mediator bradykinin,and activates TRPV1

This might suggest that this lipoxygenase product is

important in the development of inflammatory pain [22]

As TPRV1 activation in bronchial afferents leads to

bronchoconstriction and lipoxygenase activity is

up-regula-ted during inflammatory reactions,it has also been

hypo-thesized that lipoxygenase product-mediated activation of

TRPV1 might contribute to bradykinin-induced

hypersen-sitivity of airways and asthma [33] Subsequent studies

performed with TRPV1 null mice have shown,however,

that whereas TRPV1 may have a modulatory role in the

activation of bronchopulmonary C-fibres induced by

brady-kinin,it is not required for action potential discharge evoked

by this stimulus [34]

Anandamide

Biosynthesis

The endogenous lipid anandamide was isolated from pig

brain in 1992 [35] and characterized originally as an

endogenous agonist of cannabinoid receptors Therefore,

its biosynthetic and metabolic pathways have been studied

in great detail (Fig 4) [36,37] In 1999, it was reported that

anandamide could also activate rat TRPV1 receptors in

mesenteric arteries as well as both rat and human TRPV1 in

heterologous expression systems This made this compound

the first discovered endogenous ligand for TRPV1 [38,39] It

is widely recognized that anandamide is not stored in

vescicles like other mediators but by analogy with other

eicosanoids,is produced on demand in a Ca2+-dependent

manner [40] This is the result of a biosynthetic mechanism

relying on the existence of a phospholipid precursor for

anandamide,and of a Ca2+-sensitive phosphodiesterease for the conversion of this precursor into anandamide Although the biosynthetic route underlying the formation

of anandamide has been extensively studied,the phospho-lipase D (PLD) responsible for release of anandamide from its precursor N-arachidonoylphosphatidylethanolamine has only been purified recently,cloned and characterized [41]

It is still unknown which type of neurons express the biosynthetic enzymes,and therefore a classification of

anandamidergic neurons is not yet possible Nevertheless, anandamide has been found in all the brain regions which express TRPV1,with high levels in hippocampus,substantia nigra and striatum Sensory neurons of rat dorsal root ganglia are also able to produce anandamide in high amounts [42]

Degradation Theoretically,intracellular anandamide can be inactivated through two concurrent processes [43] Firstly,anandamide might be extruded from the cell via a selective transporter This might be the same protein that is responsible for the uptake of anandamide from the extracellular space into the cell,because,among other things,anandamide has been proposed to be transported by a carrier-facilitated diffusion process according to its concentration gradient across the membrane [44,45] This process might be bi-directional, because it is neither dependent on external Na+nor affected

by metabolic inhibitors Anandamide uptake (and possibly its release) can be stimulated by NO [28] and blocked by selective inhibitors [46–48] However,the elusive nature of the putative protein responsible for endocannabinoid transport across membranes has initiated a debate on its existence [12]

Secondly,once inside the cell,anandamide undergoes metabolism via two possible pathways: hydrolysis and oxygenation [43,49,50] The amide bond in anandamide can be hydrolysed,which yields AA and ethanolamine Although AA does not activate TRPV1 by itself,it is a substrate for lipoxygenases,which can produce metabolites active on this channel (see above) Therefore,when studying the effects of exogenous anandamide at TRPV1 its hydro-lysis should be taken into account Fatty acid amide hydrolase,the protein responsible for anandamide hydro-lysis in vivo,has been cloned and studied in detail [23,51] It

is an intracellular membrane-associated hydrolase with high activity in several brain areas,including the hippocampus,

Fig 4 Biosynthetic and metabolic pathways of anandamide.

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subtantia nigra and striatum [52] At the moment it is

not known whether TRPV1 is coexpressed with fatty acid

amide hydrolase in the same neurons

Much less is known about the oxygenation pathways

Lipoxygenase- and cycloxygenase-catalyzed oxygenation of

anandamide has been shown to generate a vast array of

possibly biologically active compounds,such as

hydro-peroxy-anandamides and prostamides [50] While the latter

compounds are not able to activate TRPV1,and hence

conversion of anandamide by cyclooxygenase-2 represents

an inactivation pathway,the lipoxygenase products of

anandamide might still be able to activate this channel

[29,30], much in the same way they appear to be still active

to some extent also on cannabinoid receptors [27]

Pharmacology and physiological actions

The pharmacology of anandamide actions on TRPV1 has

recently been reviewed extensively by Ross [53] In short,

anandamide displaces [3H]RTX from TPRV1 with a Kiof

 2 lMin recombinant cell lines,which is similar to that of

capsaicin [54,55] However, its potency in various assays is

usually five- to 10-fold lower than that of capsaicin For

example,in high recombinant expression systems,the EC50

value for anandamide-induced Ca2+-influx ranges from 0.4

to 5 lMand anandamide appears to act as a full agonist,

while in native systems,such as Ca2+-influx and inward

current in DRG neurons,anandamide is a partial agonist

with a potency varying from 6 to 10 lM [10,16,56] In

isolated organs,exogenous anandamide has also been

shown to induce typical TRPV1-mediated effects with

varying potency and efficacy For example,anandamide

induces CGRP-mediated relaxation of blood vessels with an

EC50of 0.3–0.8 lMas a full agonist [38,57,58], while it is

much less potent when causing tachykinin-mediated

con-striction of bronchi and urinary bladder (EC50varying from

6 to > 10 lM) [57]

Due to its low potency and partial agonism in some

assays,anandamide’s ability to be a physiologically relevant

activator of TRPV1 was originally controversial [59]

However,it is now well established that the potency and

efficacy of (exogenous) anandamide at TRPV1 receptors are

influenced by a multitude of different factors,ranging from

assay conditions and species differences to TRPV1

modi-fication and the ability of anandamide to reach the

intracellular binding site on TRPV1

Thus,due to its low intrinsic efficacy,anandamide is a

partial agonist in tissues with a low receptor reserve (e.g

bronchi),but it appears to be a full agonist in tissues with a

high receptor reserve such as the mesentery artery [53,57] In

several pathological conditions,TRPV1 activity/expression

is upregulated,and this results in a significantly higher

efficacy of anandamide [60] It has been shown that ethanol,

which is frequently used as a vehicle to dissolve

anand-amide,can potentiate TRPV1 mediated-responses to

anandamide via an unknown mechanism [61],whereas

bovine serum albumin and plastic may prevent anandamide

from reaching the intracellular binding site [55] Apart from

high temperature (> 43°C) and low pH (< 7.2),which

may activate and/or sensitize TRPV1,multiple signalling

pathways have also been shown to interact with TRPV1 to

modify its gating properties and response to anandamide

[1] The channel might be sensitized by (a) removal of its inhibition by phosphatidylinositolbisphosphate,via PLC-mediated hydrolysis [62],(b) protein kinase C-catalysed phosphorylation following PLC-mediated diacylglycerol release [63,64], (c) protein kinase A-mediated phosphoryla-tion [31],(d) phosphorylaphosphoryla-tion induced by by calmodulin-dependent protein kinase II [65] or (e) voltage-calmodulin-dependent priming [66] Conversely,TRPV1 can be rapidly desensi-tized subsequent to activating stimuli by a calmodulin-dependent step [67]

As discussed in the previous paragraph,the level of expression and the activity of the putative anandamide transporter,which may vary in different types of cells,is of crucial importance for exogenous anandamide to reach its cytosolic binding site at TRPV1 [8,57] The rapid metabo-lism of anandamide inside cells may also limit its activation

of TRPV1 [8,54,68], or potentiate its effect in the case of lipoxygenase-mediated conversion [29,30]

N-acyl-ethanolamine anandamide congeners,which are cobiosynthesized with anandamide from the phospholipase D-dependent hydrolysis of the corresponding N-acyl-phosphatidylethanolamines [41],also significantly enhance ananadamide effects at TRPV1 [55,69] One of these compounds, N-oleoylethanolamine,was even found to be able to activate TRPV1 per se under certain conditions [70] Last,but not least,anandamide functional activity at TRPV1 can be significantly masked by its concomitant activity on cannabinoid CB1receptors,particularly in those tissues and cells where the two receptors are coexpressed and are coupled to opposing biological effects [80] In this case,a significant enhancement of the potency of anand-amide at TRPV1 can be observed in the presence of CB1

antagonists [80]

At the moment,very few studies have been reported in which endogenous anandamide was shown to activate TRPV1 in vivo Although exogenous anandamide induces TRPV1-mediated vasodilation,calcium influx in DRG neurons and bronchoconstriction in situ,which might suggest that endogenous anandamide is involved through TRPV1 in physiological processes such as blood pressure, pain sensation and airway responsiveness,respectively,to date there is only little experimental data providing conclu-sive evidence supporting these hypotheses [14,60,71,72] However,recently the first example of activation of TRPV1

by endogenous anandamide has been reported Ananda-mide,endogenously produced in the inflamed ileum of rats treated with toxin A,was shown to cause TRPV1-depend-ent ileitis [68] This finding strengthens the hypothesis that anandamide behaves as an endovanilloid particularly under some pathological conditions,such as inflammation With respect to the central nervous system,it is not known whether endogenously formed anandamide activates TRPV1 under normal physiological conditions Exogenous anandamide induces,in hippocampal slices,a TRPV1-mediated enhancement of paired-pulse depression,which is

a form of short-term synaptic plasticity [73] Tonically activated TRPV1 receptors have been found in the substantia nigra compacta While the mechanism of activa-tion has not been elucidated,this suggests that TRPV1 might be involved in the control of movement [74] Interestingly,the anandamide uptake inhibitor and TRPV1 agonist,AM404

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[N-(4-hydroxyphenyl)-arachidonyl-ethanolamine],and capsaicin could attenuate,via direct

activation of TRPV1,motor disturbances by restoring

GABA and dopamine transmission in an animal model for

Huntington’s disease [75,76] However, capsazepine alone

did not affect motor disturbances,arguing against a tonic

activation of TRPV1 receptors in this animal model [76]

Finally,hippocampal anandamide has been shown to be

up-regulated during kainate-induced seizures in mice [77],

while capsazepine was shown to reduce neuronal damage in

another animal model of excitotoxicity [78] This suggests

that anandamide-mediated activation of TRPV1 might

contribute to neurotoxicity and neurodegeneration under

conditions of excitotoxicity [79]

Conclusion and perspectives

At the moment,three classes of putative endogenous ligands

for TRPV1 have been identified,i.e

N-arachidonoyldop-amine,lipoxygenase products of AA and anandamide,

thereby providing a rationale for the presence of this protein

outside the peripheral efferents of sensory neurons [5]

Although much progress has been made towards their

characterization as endovanilloids,it is still unclear whether

any of these ligands is indeed responsible of

TRPV1-mediated physiological and pathological effects in and

outside the sensory nervous system To clarify this issue,it

will be necessary to identify the physiological processes in

which TRPV1 is participating,which can then lead to the

characterization of its endogenous activators Novel

endo-vanilloids might also be discovered in this way Conversely,

knowledge about the stimulus-induced formation of any of

the three classes of the endogenous ligands can lead to the

identification of TRPV1-mediated physiological processes

By looking at the biosynthetic routes of the putative

endovanilloids,it can be suggested that each of the ligands is

formed in a spatially and temporally different manner to

selectively gate TRPV1 with different potencies and

effica-cies For example,while constitutive activity of

lipoxygen-ases responsible for the formation of some endovanilloids is

low under normal physiological conditions – especially in

the central nervous system – its activity and expression are

upregulated under pathological conditions,such as

inflam-mation,asthma and neurodegeneration At the moment,the

biosynthetic pathway of N-arachidonoyldopamine and its

regulation still need to be clarified Although anandamide

biosynthesis is also significantly upregulated during

patho-logical conditions,it is important to notice that normal

neuronal activity leads to production of anandamide,

thereby making anandamide a likely candidate to be an

endovanilloid in the central nervous system,also under

physiological conditions Molecular probes directed at

manipulating the biosynthesis and metabolism of putative

endovanilloids and genetically engineered mice lacking

functional genes for fatty acid amide hydrolase,12- or

15-lipoxygenase,

N-acyl-phosphatidylethanolamine-select-ive PLD and TRPV1,will be highly informatN-acyl-phosphatidylethanolamine-select-ive while

dissecting the physiological processes in which

endovanil-loids are active Finally,the biosynthetic and metabolic

proteins of endovanilloids might serve as valuable targets

for drug development for the increasing number of diseases

associated with abnormal TRPV1 signaling,such as

neuro-pathic pain,cough and urinary incontinence [2,71]

Acknowledgements

The authors’ research on this subject is funded by a research grant from the Volkswagen Stiftung.

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