Molecular cloning, functional characterization of the porcine transient receptor potential V1 (pTRPV1) and pharmacological comparison with endogenous pTRPV1

Ohta, Toshio; Komatsu, Ryuichi; Imagawa, Toshiaki; Otsuguro, Ken-ichi; S, Itó

doi:10.1016/j.bcp.2005.09.028

Cited by 27 publications

(21 citation statements)

References 46 publications

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“…1F) TRPV1 functions as a polymodal receptor that is activated not only by the vanilloid agonist capsaicin, but also by protons, noxious heat (2), and endovanilloids (3,4). We previously reported that these stimuli are effective for activating pTRPV1 (16). Next we examined whether low [Na ϩ ] o produced potentiation effects on proton-, heat-, or endovanilloid-induced .…”

Section: Methodsmentioning

confidence: 99%

“…Human embryonic kidney (HEK) 293 cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin G (Meiji-Seika), and 100 g/ml streptomycin (Banyu). The expression vectors pcDNA-pTRPV1-DEST53 for N-terminal GFP fusion protein, pcDNA-pTRPV1-DEST47 for C-terminal GFP fusion protein, and pGATE-pTRPV1 were constructed using Gateway Technology (Invitrogen) as described previously (16). Cells were transfected with the expression vector using a transfection reagent (Lipofectamine 2000, Invitrogen) and used 24 -48 h after transfection.…”

Section: Methodsmentioning

confidence: 99%

“…All clones and mutations used were verified by sequencing using an automated sequencer (AB1 PRISM 3100, Applied Biosystems). Porcine dorsal root ganglion (DRG) neurons were obtained from male pigs (2-4 weeks after birth, 5-10 kg) as described previously (16). In brief, pigs were deeply anesthetized by sodium pentobarbital (50 mg/kg, intraperitoneal) after tranquilization with ketamine (15 mg/kg, intramascular) and then killed by bloodletting from the cervical artery.…”

Section: Methodsmentioning

confidence: 99%

“…We recently cloned a porcine orthologue of TRPV1 (pTRPV1) and analyzed its functional properties using a heterologous expression system (16). We showed that pTRPV1 was a nonselective cation channel sensitive to a number of vanilloid agonists, including capsaicin and endovanilloids, heat, and protons.…”

mentioning

confidence: 99%

“…Phorbol 12-phenylacetate 13-acetate 20-homovanillate (PPAHV), a TRPV1 agonist, can stimulate rodent (19 -22) but not human TRPV1 (20). Capsazepine, a TRPV1 antagonist, does not antagonize proton-and heat-induced responses in rodent (20,21) but does antagonize in human (20) and porcine TRPV1 (16). Therefore, using a porcine orthologue may be more advantageous to predict physiological and pharmacological characteristics in human TRPV1.…”

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confidence: 99%

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Novel Gating and Sensitizing Mechanism of Capsaicin Receptor (TRPV1)

Ohta

Imagawa

2008

Journal of Biological Chemistry

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Transient receptor potential V1 (TRPV1) is a nonselective cation channel expressed in nociceptors andsurrounding several proton-sensitive sites on the extracellular side of the pore-forming loop of the TRPV1 channel may play an important role as a brake to suppress the excessive activity of this channel under physiological conditions. Transient receptor potential V1 (TRPV1) 2 is a nonselective cation channel with high Ca 2ϩ permeability found in sensory neurons that functions as a molecular integrator of pain perception (1). TRPV1 is activated by polymodal stimuli such as capsaicin, protons, and noxious heat (2). Some endogenous ligands termed endovanilloids (3), including 15-hydroperoxyeicosa-5Z,8Z,11Z,13E-tetraenoic acid (15(S)-HPETE), a lipoxygenase product, also activate TRPV1 (4). Because mice genetically lacking the TRPV1 channel exhibit impaired nociception (5, 6) and several TRPV1 antagonists have antinociceptive activity in vivo (7-9), TRPV1 is considered to be a key component of signal transduction pathways in the nociceptive system. Several regulatory mechanisms of TRPV1 function have been elucidated. Sensitization can arise from phosphorylation of TRPV1 at multiple sites via protein kinase C and protein kinase A downstream of activation of GTP-binding protein-coupling receptor by bradykinin, ATP, nerve growth factor, calcitonin gene-related peptide, and prostaglandins (10). Our recent report also showed that the metabotropic 5-HT receptor facilitates TRPV1 functions through protein kinase C/protein kinase A pathways (11). TRPV1 is subject to tonic inhibition by phosphatidylinositol 4,5-bisphosphate (PIP 2 ) (12), providing an additional biochemical pathway through which the activity of the channel can be regulated after GTPbinding protein-coupling receptor activation. Recently, increases of cationic strength have been shown to contribute to inflammatory pain signaling through modulation of TRPV1 channels, since extracellular cations such as Na ϩ , Mg 2ϩ , and Ca 2ϩ gate and sensitize them (13). Hyponatremia, a disease due to reduction of the plasma Na ϩ concentration, is one of the most common electrolyte disorders (14). It occurs via excess loss of plasma Na ϩ as a result of various causes such as chronic renal failure and congestive heart failure. When the serum Na ϩ level falls gradually over a period of several days or weeks, sodium levels as low as 110 meq/liter may be reached with minimal symptomatology. Its symptoms are often concomitant with pain (15). However, the molecular detector of pain perception in hyponatremia has not yet been identified.We recently cloned a porcine orthologue of TRPV1 (pTRPV1) and analyzed its functional properties using a heterologous expression system (16). We showed that pTRPV1 was a nonselective cation channel sensitive to a number of vanilloid agonists, including capsaicin and endovanilloids, heat, and protons. In pharmacological characteristics of TRPV1, there are remarkable species differences; chicken TRPV1 is not sensitive * This work was supported by grants...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Novel Gating and Sensitizing Mechanism of Capsaicin Receptor (TRPV1)

Ohta

Imagawa

2008

Journal of Biological Chemistry

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TRPV1 Genetics

Dorfman

Tsui

Salter

et al. 2010

Vanilloid Receptor TRPV1 in Drug Discovery

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Neurochemical features of boar lumbosacral dorsal root ganglion neurons and characterization of sensory neurons innervating the urinary bladder trigone

Russo

Clavenzani

Sorteni

et al. 2012

J of Comparative Neurology

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Porcine lumbosacral dorsal root ganglion (DRG) neurons were neurochemically characterized by using six neuronal markers: calcitonin gene‐related peptide (CGRP), substance P (SP), neuronal nitric oxide synthase (nNOS), neurofilament 200kDa (NF200), transient receptor potential vanilloid 1 (TRPV1), and isolectin B4 (IB4) from Griffonia simplicifolia. In addition, the phenotype and cross‐sectional area of DRG neurons innervating the urinary bladder trigone (UBT) were evaluated by coupling retrograde tracer technique and immunohistochemistry. Lumbar and sacral DRG neuronal subpopulations were immunoreactive (IR) for CGRP (30 ± 3% and 29 ± 3%, respectively), SP (26 ± 8% and 27 ± 12%, respectively), nNOS (21 ± 4% and 26 ± 7%, respectively), NF200 (75 ± 14% and 81 ± 7%, respectively), and TRPV1 (48 ± 13% and 43 ± 6%, respectively), and labeled for IB4 (56 ± 6% and 43 ± 10%, respectively). UBT sensory neurons, which were distributed from L2 to Ca1 DRG, had a segmental localization, showing their highest density in L4–L5 and S2–S4 DRG. Lumbar and sacral UBT sensory neurons expressed similar percentages of NF200 immunoreactivity (64 ± 33% and 58 ± 12%, respectively) but showed a significantly different immunoreactivity for CGRP, SP, nNOS, and TRPV1 (56 ± 9%, 39 ± 15%, 17 ± 13%, 62 ± 10% vs. 16 ± 6%, 16 ± 11%, 6 ± 1%, 45 ± 24%, respectively). Lumbar and sacral UBT sensory neurons also showed different IB4 labeling (67 ± 19% and 48 ± 16, respectively). Taken together, these data indicate that the lumbar and sacral pathways probably play different roles in sensory transmission from the UBT. The findings related to cell size also reinforced this hypothesis, because lumbar UBT sensory neurons were significantly larger than sacral ones (1,112 ± 624 μm2 vs. 716 ± 421 μm2). J. Comp. Neurol. 521:342–366, 2013. © 2012 Wiley Periodicals, Inc.

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Molecular cloning, functional characterization of the porcine transient receptor potential V1 (pTRPV1) and pharmacological comparison with endogenous pTRPV1

Cited by 27 publications

References 46 publications

Novel Gating and Sensitizing Mechanism of Capsaicin Receptor (TRPV1)

Novel Gating and Sensitizing Mechanism of Capsaicin Receptor (TRPV1)

TRPV1 Genetics

Neurochemical features of boar lumbosacral dorsal root ganglion neurons and characterization of sensory neurons innervating the urinary bladder trigone

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